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Sample records for microbial biomass enzyme

  1. Phosphorus fractions, microbial biomass and enzyme activities in ...

    African Journals Online (AJOL)

    Potohar, northern Punjab, Pakistan in September, 2008 and analysed for P fractions and microbial parameters including microbial biomass C, microbial biomass N, microbial biomass P, and activities of dehydrogenase and alkaline phosphatase enzymes. The average size of different P fractions (% of total P) in the soils ...

  2. Effects of uranium on soil microbial biomass carbon, enzymes, plant biomass and microbial diversity in yellow soils

    International Nuclear Information System (INIS)

    Yan, X.; Zhang, Y.; Luo, X.; Yu, L.

    2016-01-01

    We conducted an experiment to investigate the effects of uranium (U) on soil microbial biomass carbon (MBC), enzymes, plant biomass and microbial diversity in yellow soils under three concentrations: 0 mg kg"-"1 (T1, control), 30 mg kg"-"1 (T2) and 60 mg kg"-"1 (T3). Under each treatment, elevated U did not reduce soil MBC or plant biomass, but inhibited the activity of the soil enzymes urease (UR), dehydrogenase (DH) and phosphatase (PHO). The microbial diversity was different, with eight dominant phyla in T1 and six in T2 and T3. Furthermore, Proteobacteria and material X were both detected in each treatment site (T1, T2 and T3). Pseudomonas sp. was the dominant strain, followed by Acidiphilium sp. This initial study provided valuable data for further research toward a better understanding of U contamination in yellow soils in China. (authors)

  3. [Effects of altitudes on soil microbial biomass and enzyme activity in alpine-gorge regions.

    Science.gov (United States)

    Cao, Rui; Wu, Fu Zhong; Yang, Wan Qin; Xu, Zhen Feng; Tani, Bo; Wang, Bin; Li, Jun; Chang, Chen Hui

    2016-04-22

    In order to understand the variations of soil microbial biomass and soil enzyme activities with the change of altitude, a field incubation was conducted in dry valley, ecotone between dry valley and mountain forest, subalpine coniferous forest, alpine forest and alpine meadow from 1563 m to 3994 m of altitude in the alpine-gorge region of western Sichuan. The microbial biomass carbon and nitrogen, and the activities of invertase, urease and acid phosphorus were measured in both soil organic layer and mineral soil layer. Both the soil microbial biomass and soil enzyme activities showed the similar tendency in soil organic layer. They increased from 2158 m to 3028 m, then decreased to the lowest value at 3593 m, and thereafter increased until 3994 m in the alpine-gorge region. In contrast, the soil microbial biomass and soil enzyme activities in mineral soil layer showed the trends as, the subalpine forest at 3028 m > alpine meadow at 3994 m > montane forest ecotone at 2158 m > alpine forest at 3593 m > dry valley at 1563 m. Regardless of altitudes, soil microbial biomass and soil enzyme activities were significantly higher in soil organic layer than in mineral soil layer. The soil microbial biomass was significantly positively correlated with the activities of the measured soil enzymes. Moreover, both the soil microbial biomass and soil enzyme activities were significantly positively correlated with soil water content, organic carbon, and total nitrogen. The activity of soil invertase was significantly positively correlated with soil phosphorus content, and the soil acid phosphatase was so with soil phosphorus content and soil temperature. In brief, changes in vegetation and other environmental factors resulting from altitude change might have strong effects on soil biochemical properties in the alpine-gorge region.

  4. [Effects of bio-crust on soil microbial biomass and enzyme activities in copper mine tailings].

    Science.gov (United States)

    Chen, Zheng; Yang, Gui-de; Sun, Qing-ye

    2009-09-01

    Bio-crust is the initial stage of natural primary succession in copper mine tailings. With the Yangshanchong and Tongguanshan copper mine tailings in Tongling City of Anhui Province as test objects, this paper studied the soil microbial biomass C and N and the activities of dehydrogenase, catalase, alkaline phosphatase, and urease under different types of bio-crust. The bio-crusts improved the soil microbial biomass and enzyme activities in the upper layer of the tailings markedly. Algal crust had the best effect in improving soil microbial biomass C and N, followed by moss-algal crust, and moss crust. Soil microflora also varied with the type of bio-crust. No'significant difference was observed in the soil enzyme activities under the three types of bio-crust. Soil alkaline phosphatase activity was significantly positively correlated with soil microbial biomass and dehydrogenase and urease activities, but negatively correlated with soil pH. In addition, moss rhizoid could markedly enhance the soil microbial biomass and enzyme activities in moss crust rhizoid.

  5. Changes in Soil Enzyme Activities and Microbial Biomass after Revegetation in the Three Gorges Reservoir, China

    Directory of Open Access Journals (Sweden)

    Qingshui Ren

    2018-05-01

    Full Text Available Soil enzymes and microbes are central to the decomposition of plant and microbial detritus, and play important roles in carbon, nitrogen, and phosphorus biogeochemistry cycling at the ecosystem level. In the present study, we characterized the soil enzyme activity and microbial biomass in revegetated (with Taxodium distichum (L. Rich. and Cynodon dactylon (L. Pers. versus unplanted soil in the riparian zone of the Three Gorges Dam Reservoir (TGDR, in order to quantify the effect of revegetation on the edaphic microenvironment after water flooding in situ. After revegetation, the soil physical and chemical properties in revegetated soil showed significant differences to those in unplanted soil. The microbial biomass carbon and phosphorus in soils of T. distichum were significantly higher than those in C. dactylon and unplanted soils, respectively. The microbial biomass nitrogen in revegetated T. distichum and C. dactylon soils was significantly increased by 273% and 203%, respectively. The enzyme activities of T. distichum and C. dactylon soils displayed no significant difference between each other, but exhibited a great increase compared to those of the unplanted soil. Elements ratio (except C/N (S did not vary significantly between T. distichum and C. dactylon soils; meanwhile, a strong community-level elemental homeostasis in the revegetated soils was found. The correlation analyses demonstrated that only microbial biomass carbon and phosphorus had a significantly positive relationship with soil enzyme activities. After revegetation, both soil enzyme activities and microbial biomasses were relatively stable in the T. distichum and C. dactylon soils, with the wooded soil being more superior. The higher enzyme activities and microbial biomasses demonstrate the C, N, and P cycling and the maintenance of soil quality in the riparian zone of the TGDR.

  6. Dynamic relationships between microbial biomass, respiration, inorganic nutrients and enzyme activities: informing enzyme based decomposition models

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    Daryl L Moorhead

    2013-08-01

    Full Text Available We re-examined data from a recent litter decay study to determine if additional insights could be gained to inform decomposition modeling. Rinkes et al. (2013 conducted 14-day laboratory incubations of sugar maple (Acer saccharum or white oak (Quercus alba leaves, mixed with sand (0.4% organic C content or loam (4.1% organic C. They measured microbial biomass C, carbon dioxide efflux, soil ammonium, nitrate, and phosphate concentrations, and β-glucosidase (BG, β-N-acetyl-glucosaminidase (NAG, and acid phosphatase (AP activities on days 1, 3, and 14. Analyses of relationships among variables yielded different insights than original analyses of individual variables. For example, although respiration rates per g soil were higher for loam than sand, rates per g soil C were actually higher for sand than loam, and rates per g microbial C showed little difference between treatments. Microbial biomass C peaked on day 3 when biomass-specific activities of enzymes were lowest, suggesting uptake of litter C without extracellular hydrolysis. This result refuted a common model assumption that all enzyme production is constitutive and thus proportional to biomass, and/or indicated that part of litter decay is independent of enzyme activity. The length and angle of vectors defined by ratios of enzyme activities (BG/NAG versus BG/AP represent relative microbial investments in C (length, and N and P (angle acquiring enzymes. Shorter lengths on day 3 suggested low C limitation, whereas greater lengths on day 14 suggested an increase in C limitation with decay. The soils and litter in this study generally had stronger P limitation (angles > 45˚. Reductions in vector angles to < 45˚ for sand by day 14 suggested a shift to N limitation. These relational variables inform enzyme-based models, and are usually much less ambiguous when obtained from a single study in which measurements were made on the same samples than when extrapolated from separate studies.

  7. Field and lab conditions alter microbial enzyme and biomass dynamics driving decomposition of the same leaf litter.

    Science.gov (United States)

    Rinkes, Zachary L; Sinsabaugh, Robert L; Moorhead, Daryl L; Grandy, A Stuart; Weintraub, Michael N

    2013-01-01

    Fluctuations in climate and edaphic factors influence field decomposition rates and preclude a complete understanding of how microbial communities respond to plant litter quality. In contrast, laboratory microcosms isolate the intrinsic effects of litter chemistry and microbial community from extrinsic effects of environmental variation. Used together, these paired approaches provide mechanistic insights to decomposition processes. In order to elucidate the microbial mechanisms underlying how environmental conditions alter the trajectory of decay, we characterized microbial biomass, respiration, enzyme activities, and nutrient dynamics during early (40% mass loss) decay in parallel field and laboratory litter bag incubations for deciduous tree litters with varying recalcitrance (dogwood litter types, despite above-freezing soil temperatures and adequate moisture during these winter months. In contrast, microcosms displayed high C mineralization rates in the first week. During mid-decay, the labile dogwood and maple litters in the field had higher mass loss per unit enzyme activity than the lab, possibly due to leaching of soluble compounds. Microbial biomass to litter mass (B:C) ratios peaked in the field during late decay, but B:C ratios declined between mid- and late decay in the lab. Thus, microbial biomass did not have a consistent relationship with litter quality between studies. Higher oxidative enzyme activities in oak litters in the field, and higher nitrogen (N) accumulation in the lab microcosms occurred in late decay. We speculate that elevated N suppressed fungal activity and/or biomass in microcosms. Our results suggest that differences in microbial biomass and enzyme dynamics alter the decay trajectory of the same leaf litter under field and lab conditions.

  8. Tracking dynamics of plant biomass composting by changes in substrate structure, microbial community, and enzyme activity

    Science.gov (United States)

    2012-01-01

    Background Understanding the dynamics of the microbial communities that, along with their secreted enzymes, are involved in the natural process of biomass composting may hold the key to breaking the major bottleneck in biomass-to-biofuels conversion technology, which is the still-costly deconstruction of polymeric biomass carbohydrates to fermentable sugars. However, the complexity of both the structure of plant biomass and its counterpart microbial degradation communities makes it difficult to investigate the composting process. Results In this study, a composter was set up with a mix of yellow poplar (Liriodendron tulipifera) wood-chips and mown lawn grass clippings (85:15 in dry-weight) and used as a model system. The microbial rDNA abundance data obtained from analyzing weekly-withdrawn composted samples suggested population-shifts from bacteria-dominated to fungus-dominated communities. Further analyses by an array of optical microscopic, transcriptional and enzyme-activity techniques yielded correlated results, suggesting that such population shifts occurred along with early removal of hemicellulose followed by attack on the consequently uncovered cellulose as the composting progressed. Conclusion The observed shifts in dominance by representative microbial groups, along with the observed different patterns in the gene expression and enzymatic activities between cellulases, hemicellulases, and ligninases during the composting process, provide new perspectives for biomass-derived biotechnology such as consolidated bioprocessing (CBP) and solid-state fermentation for the production of cellulolytic enzymes and biofuels. PMID:22490508

  9. Field and lab conditions alter microbial enzyme and biomass dynamics driving decomposition of the same leaf litter

    Directory of Open Access Journals (Sweden)

    Zachary L Rinkes

    2013-09-01

    Full Text Available Fluctuations in climate and edaphic factors influence field decomposition rates and preclude a complete understanding of how microbial communities respond to plant litter quality. In contrast, laboratory microcosms isolate the intrinsic effects of litter chemistry and microbial community from extrinsic effects of environmental variation. Used together, these paired approaches provide mechanistic insights to decomposition processes. In order to elucidate the microbial mechanisms underlying how environmental conditions alter the trajectory of decay, we characterized microbial biomass, respiration, enzyme activities, and nutrient dynamics during early (< 10% mass loss, mid- (10-40% mass loss, and late (> 40% mass loss decay in parallel field and laboratory litter bag incubations for deciduous tree litters with varying recalcitrance (dogwood < maple < maple-oak mixture < oak. In the field, mass loss was minimal (< 10% over the first 50 days (January-February, even for labile litter types, despite above-freezing soil temperatures and adequate moisture during these winter months. In contrast, microcosms displayed high C mineralization rates in the first week. During mid-decay, the labile dogwood and maple litters in the field had higher mass loss per unit enzyme activity than the lab, possibly due to leaching of soluble compounds. Microbial biomass to litter mass (B:C ratios peaked in the field during late decay, but B:C ratios declined between mid- and late decay in the lab. Thus, microbial biomass did not have a consistent relationship with litter quality between studies. Higher oxidative enzyme activities in oak litters in the field, and higher nitrogen (N accumulation in the lab microcosms occurred in late decay. We speculate that elevated N suppressed fungal activity and/or biomass in microcosms. Our results suggest that differences in microbial biomass and enzyme dynamics alter the decay trajectory of the same leaf litter under field and lab

  10. Tracking dynamics of plant biomass composting by changes in substrate structure, microbial community, and enzyme activity

    Directory of Open Access Journals (Sweden)

    Wei Hui

    2012-04-01

    Full Text Available Abstract Background Understanding the dynamics of the microbial communities that, along with their secreted enzymes, are involved in the natural process of biomass composting may hold the key to breaking the major bottleneck in biomass-to-biofuels conversion technology, which is the still-costly deconstruction of polymeric biomass carbohydrates to fermentable sugars. However, the complexity of both the structure of plant biomass and its counterpart microbial degradation communities makes it difficult to investigate the composting process. Results In this study, a composter was set up with a mix of yellow poplar (Liriodendron tulipifera wood-chips and mown lawn grass clippings (85:15 in dry-weight and used as a model system. The microbial rDNA abundance data obtained from analyzing weekly-withdrawn composted samples suggested population-shifts from bacteria-dominated to fungus-dominated communities. Further analyses by an array of optical microscopic, transcriptional and enzyme-activity techniques yielded correlated results, suggesting that such population shifts occurred along with early removal of hemicellulose followed by attack on the consequently uncovered cellulose as the composting progressed. Conclusion The observed shifts in dominance by representative microbial groups, along with the observed different patterns in the gene expression and enzymatic activities between cellulases, hemicellulases, and ligninases during the composting process, provide new perspectives for biomass-derived biotechnology such as consolidated bioprocessing (CBP and solid-state fermentation for the production of cellulolytic enzymes and biofuels.

  11. The effect of D123 wheat as a companion crop on soil enzyme activities, microbial biomass and microbial communities in the rhizosphere of watermelon.

    Science.gov (United States)

    Xu, Weihui; Wang, Zhigang; Wu, Fengzhi

    2015-01-01

    The growth of watermelon is often threatened by Fusarium oxysporum f. sp. niveum (Fon) in successively monocultured soil, which results in economic loss. The objective of this study was to investigate the effect of D123 wheat as a companion crop on soil enzyme activities, microbial biomass and microbial communities in the rhizosphere of watermelon and to explore the relationship between the effect and the incidence of wilt caused by Fon. The results showed that the activities of soil polyphenol oxidase, urease and invertase were increased, the microbial biomass nitrogen (MBN) and microbial biomass phosphorus (MBP) were significantly increased, and the ratio of MBC/MBN was decreased (P Fusarium wilt was also decreased in the watermelon/wheat companion system. In conclusion, this study indicated that D123 wheat as a companion crop increased soil enzyme activities and microbial biomass, decreased the Fon population, and changed the relative abundance of microbial communities in the rhizosphere of watermelon, which may be related to the reduction of Fusarium wilt in the watermelon/wheat companion system.

  12. The effect of D123 wheat as a companion crop on soil enzyme activities, microbial biomass and microbial communities in the rhizosphere of watermelon

    Directory of Open Access Journals (Sweden)

    Wei Hui Xu

    2015-09-01

    Full Text Available The growth of watermelon is often threatened by Fusarium oxysporum f. sp. niveum (Fon in successively monocultured soil, which results in economic loss. The objective of this study was to investigate the effect of D123 wheat as a companion crop on soil enzyme activities, microbial biomass and microbial communities in the rhizosphere of watermelon and to explore the relationship between the effect and the incidence of wilt caused by Fon. The results showed that the activities of soil polyphenol oxidase, urease and invertase were increased, the microbial biomass nitrogen (MBN and microbial biomass phosphorus (MBP were significantly increased, and the ratio of MBC/MBN was decreased (P<0.05. Real-time PCR analysis showed that the Fon population declined significantly in the watermelon/wheat companion system compared with the monoculture system (P<0.05. The analysis of microbial communities showed that the relative abundance of microbial communities was changed in the rhizosphere of watermelon. Compared with the monoculture system, the relative abundances of Alphaproteobacteria, Actinobacteria, Gemmatimonadetes and Sordariomycetes were increased, and the relative abundances of Gammaproteobacteria, Sphingobacteria, Cytophagia, Pezizomycetes, and Eurotiomycetes were decreased in the rhizosphere of watermelon in the watermelon/wheat companion system; importantly, the incidence of Fusarium wilt was also decreased in the watermelon/wheat companion system. In conclusion, this study indicated that D123 wheat as a companion crop increased soil enzyme activities and microbial biomass, decreased the Fon population, and changed the relative abundance of microbial communities in the rhizosphere of watermelon, which may be related to the reduction of Fusarium wilt in the watermelon/wheat companion system.

  13. Conditioning biomass for microbial growth

    Science.gov (United States)

    Bodie, Elizabeth A; England, George

    2015-03-31

    The present invention relates to methods for improving the yield of microbial processes that use lignocellulose biomass as a nutrient source. The methods comprise conditioning a composition comprising lignocellulose biomass with an enzyme composition that comprises a phenol oxidizing enzyme. The conditioned composition can support a higher rate of growth of microorganisms in a process. In one embodiment, a laccase composition is used to condition lignocellulose biomass derived from non-woody plants, such as corn and sugar cane. The invention also encompasses methods for culturing microorganisms that are sensitive to inhibitory compounds in lignocellulose biomass. The invention further provides methods of making a product by culturing the production microorganisms in conditioned lignocellulose biomass.

  14. Microbial biomass carbon and enzyme activities of urban soils in Beijing.

    Science.gov (United States)

    Wang, Meie; Markert, Bernd; Shen, Wenming; Chen, Weiping; Peng, Chi; Ouyang, Zhiyun

    2011-07-01

    To promote rational and sustainable use of soil resources and to maintain the urban soil quality, it is essential to assess urban ecosystem health. In this study, the microbiological properties of urban soils in Beijing and their spatial distribution patterns across the city were evaluated based on measurements of microbial biomass carbon and urease and invertase activities of the soils for the purpose of assessing the urban ecosystem health of Beijing. Grid sampling design, normal Kriging technique, and the multiple comparisons among different land use types were used in soil sampling and data treatment. The inherent chemical characteristics of urban soils in Beijing, e.g., soil pH, electronic conductivity, heavy metal contents, total N, P and K contents, and soil organic matter contents were detected. The size and diversity of microbial community and the extent of microbial activity in Beijing urban soils were measured as the microbial biomass carbon content and the ratio of microbial biomass carbon content to total soil organic carbon. The microbial community health measured in terms of microbial biomass carbon, urease, and invertase activities varied with the organic substrate and nutrient contents of the soils and were not adversely affected by the presence of heavy metals at p urban soils influenced the nature and activities of the microbial communities.

  15. Temporal variations in microbial biomass C and cellulolytic enzyme activity in arable soils: effects of organic matter input

    DEFF Research Database (Denmark)

    Debosz, K.; Rasmussen, Peter Have; Pedersen, A. R.

    1999-01-01

    Temporal variations in soil microbial biomass C concentration and in activity of extracellular enzymes of the cellulolytic complex were investigated in a field experiment after eight years of cultivation with either low organic matter input (low-OM) or high organic matter input (high-OM). The cul......Temporal variations in soil microbial biomass C concentration and in activity of extracellular enzymes of the cellulolytic complex were investigated in a field experiment after eight years of cultivation with either low organic matter input (low-OM) or high organic matter input (high......-OM). The cultivation systems differed in whether their source of fertiliser was mainly mineral or organic, in whether a winter cover crop was grown, and whether straw was mulched or removed. Sampling occurred at approximately monthly intervals, over a period of two years. Distinct temporal variations in microbial......) and an endocellulase activity of 44.2 +/- 1.1 nmol g(-1) h(-1). (C) 1999 Elsevier Science B.V. All rights reserved....

  16. Tillage and manure effect on soil microbial biomass and respiration ...

    African Journals Online (AJOL)

    The objective of this study was to determine the influence of both tillage and liquid pig manure application on soil microbial biomass, enzyme activities and microbial respiration in a meadow soil. The results obtained did not show any significant effect of tillage and manure on microbial biomass carbon (C) and nitrogen (N) ...

  17. Effects of Nitrogen and Water on Soil Enzyme Activity and Soil Microbial Biomass in Stipa baicalensis Steppe,Inner Mongolia of North China

    Directory of Open Access Journals (Sweden)

    WANG Jie

    2014-06-01

    Full Text Available In this paper, eight nitrogen treatments were applied at 0 g·m -2(N0, 1.5 g·m -2(N15, 3.0 g·m -2(N30, 5.0 g·m -2(N50, 10.0 g·m -2(N100, 15.0 g·m -2(N150, 20.0 g·m -2(N200, 30.0 g·m -2(N300 as NH 4 NO 3 and adding water to simulate summer rainfall of 100 mm, the interactive experiment was set to explore the effects of nitrogen and water addition in Stipa baicalensis steppe on soil nutrients, enzyme activities and soil microbial biomass. The results showed that the nitrogen and water addition changed soil physico-chemical factors obviously, the content of soil total organic carbon, total nitrogen, nitrate nitrogen and ammonium nitrogen increased along with the increasing of application rate of nitrogen, on the contrary, the soil pH value had decreasing trend. Appropriate application of nitrogen could enhance the activity of urease and catalase but decreased the activity of polyphenol oxidase. Nitrogen and water addition had significant effect on soil microbial biomass C and N. Higher level of N fertilizer significantly reduced microbial biomass C, and the microbial biomass N was on the rise with the application rate of nitrogen. The addition of water could slow the inhibition of nitrogen to microorganism and increase the microbial biomass C and N. A closed relationship existed in soil nutrient, activities of soil enzyme and soil microbial biomass C and N. The significantly positive correlation existed between total N, organic C, nitrate N and catalase, significantly negative correlation between nitrate N, ammonium N, total N and polyphenol oxidase. Microbial biomass N was significantly positive correlated with total N, nitrate N, ammonium N, catalase, phosphatase, and was negative correlated with polyphenol oxidase. Microbial biomass C was significantly positive correlated with polyphenol oxidase, and was negative correlated with catalase.

  18. Soil Microbial Biomass, Basal Respiration and Enzyme Activity of Main Forest Types in the Qinling Mountains

    Science.gov (United States)

    Cheng, Fei; Peng, Xiaobang; Zhao, Peng; Yuan, Jie; Zhong, Chonggao; Cheng, Yalong; Cui, Cui; Zhang, Shuoxin

    2013-01-01

    Different forest types exert essential impacts on soil physical-chemical characteristics by dominant tree species producing diverse litters and root exudates, thereby further regulating size and activity of soil microbial communities. However, the study accuracy is usually restricted by differences in climate, soil type and forest age. Our objective is to precisely quantify soil microbial biomass, basal respiration and enzyme activity of five natural secondary forest (NSF) types with the same stand age and soil type in a small climate region and to evaluate relationship between soil microbial and physical-chemical characters. We determined soil physical-chemical indices and used the chloroform fumigation-extraction method, alkali absorption method and titration or colorimetry to obtain the microbial data. Our results showed that soil physical-chemical characters remarkably differed among the NSFs. Microbial biomass carbon (Cmic) was the highest in wilson spruce soils, while microbial biomass nitrogen (Nmic) was the highest in sharptooth oak soils. Moreover, the highest basal respiration was found in the spruce soils, but mixed, Chinese pine and spruce stands exhibited a higher soil qCO2. The spruce soils had the highest Cmic/Nmic ratio, the greatest Nmic/TN and Cmic/Corg ratios were found in the oak soils. Additionally, the spruce soils had the maximum invertase activity and the minimum urease and catalase activities, but the maximum urease and catalase activities were found in the mixed stand. The Pearson correlation and principle component analyses revealed that the soils of spruce and oak stands obviously discriminated from other NSFs, whereas the others were similar. This suggested that the forest types affected soil microbial properties significantly due to differences in soil physical-chemical features. PMID:23840671

  19. Enzyme activities and microbial biomass in topsoil layer during spontaneous succession in spoil heaps after brown coal mining

    Czech Academy of Sciences Publication Activity Database

    Baldrian, Petr; Trögl, Josef; Frouz, Jan; Šnajdr, Jaroslav; Valášková, Vendula; Merhautová, Věra; Cajthaml, Tomáš; Herinková, Jana

    2008-01-01

    Roč. 40, č. 9 (2008), s. 2107-2115 ISSN 0038-0717 R&D Projects: GA MŠk LC06066 Institutional research plan: CEZ:AV0Z50200510; CEZ:AV0Z60660521 Keywords : enzyme activity * lignocellulose * microbial biomass Subject RIV: EH - Ecology, Behaviour Impact factor: 2.926, year: 2008

  20. Soil microbial biomass in an agroforestry system of Northeast Brazil

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    Rosane C. Rodrigues

    2015-01-01

    Full Text Available Agroforestry systems (AFS are considered alternative land use options to help prevent soil degradation and improve soil microbial biomass and organic C status. However, it is unclear how different densities of babassu palm [Attalea speciosa (syn. Orbignya phalerata], which is an important tree in Northeast Brazil, affect the soil microbial biomass. We investigated the soil microbial biomass C and activity under AFS with different densities of babassu palm associated with Brachiaria brizantha grass. Soil microbial biomass C (MBC, soil microbial biomass N (MBN, MBC:total organic C ratio, fluorescein diacetate hydrolysis and dehydrogenase activity showed highest values in plots with high density of babassu palm. On the other hand, the respiratory quotient (qCO2 was significantly greater in plots without babassu palm. Brachiaria brizantha in monoculture may promote C losses from the soil, but AFS with high density of babassu palm may increase the potential of soils to accumulate C.Keywords: Enzyme activity, tropical soil, babassu palm, silvopastoral system, soil quality.DOI: 10.17138/TGFT(341-48

  1. Effect of microbial enzyme allocation strategies on stoichiometry of soil organic matter (SOM) decomposition

    Science.gov (United States)

    Wutzler, Thomas

    2014-05-01

    We explored different strategies of soil microbial community to invest resources into extracellular enzymes by conceptual modelling. Similar to the EEZY model by Moorhead et al. (2012), microbial community can invest into two separate pools of enzymes that depolymerize two different SOM pools. We show that with assuming that a fixed fraction of substrate uptake is allocated to enzymes, the microbial dynamics decouples from decomposition dynamics. We propose an alternative formulation where investment into enzymes is proportional to microbial biomass. Next, we show that the strategy of optimizing stoichiometry of decomposition flux according to microbial biomass stoichiometry yield less microbial growth than the strategy of optimizing revenue of the currently limiting element. However, both strategies result in better usage of the resources, i.e. less C overflow or N mineralization, than the strategy of equal allocation to both enzymes. Further, we discuss effects of those strategies on decomposition of SOM and priming at different time scales and discuss several abstractions from the detailed model dynamics for usage in larger scale models.

  2. [Influence of different slope position and profile in Disporopsis pernyi forest land on soil microbial biomass and enzyme activity in southwest Karst mountain of China ].

    Science.gov (United States)

    Qin, Hua-Jun; He, Bing-Hui; Zhao, Xuan-chi; Li, Yuan; Mao, Wen-tao; Zeng, Qing-ping

    2014-09-01

    Soil microbial biomass and enzyme activity are important parameters to evaluate the quality of the soil environment. The goal of this study was to determine the influence of different slope position and section in Disporopsis pernyi forest land on the soil microbial biomass and enzyme activity in southwest Karst Mountain. In this study, we chose the Dip forest land at Yunfo village Chengdong town Liangping country Chongqing Province as the study object, to analyze the influence of three different slope positions [Up Slope(US), Middle Slope(MS), Below Slope(BS)] and two different sections-upper layer(0-15 cm) and bottom layer(15-30 cm) on the soil microbial biomass carbon (SMBC), soil microbial biomass nitrogen (SMBN), microbial carbon entropy (qMBC), microbial nitrogen entropy (qMBN) , catalase(CAT), alkaline phosphatase (ALK), urease(URE), and invertase(INV). The results showed that the same trend (BS > MS > US) was found for SMBC, SMBN, qMBC, qMBN, CAT and INV of upper soil layer, while a different trend (BS > US > MS) was observed for ALK. In addition, another trend (MS > US > BS) was observed for URE. The same trend (BS > MS >US) was observed for SMBN, qMBN, CAT, ALK, URE and INV in bottom layer, but a different trend (MS > BS > US) was observed for SMBC and qMBC. The SMBC, SMBN, CAT, ALK, URE and INV manifested as upper > bottom with reduction of the section, while qMBC and qMBN showed the opposite trend. Correlation analysis indicated that there were significant (P <0.05) or highly significant (P < 0.01) positive correlations among SMBC in different slope position and section, soil enzyme activity and moisture. According to the two equations of regression analysis, SMBC tended to increase with the increasing CAT and ALK, while decreased with the increasing pH. Then SMBN tended to increase with the increasing URE and INV.

  3. Contrasting effects of ammonium and nitrate additions on the biomass of soil microbial communities and enzyme activities in subtropical China

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    C. Zhang

    2017-10-01

    Full Text Available The nitrate to ammonium ratios in nitrogen (N compounds in wet atmospheric deposits have increased over the recent past, which is a cause for some concern as the individual effects of nitrate and ammonium deposition on the biomass of different soil microbial communities and enzyme activities are still poorly defined. We established a field experiment and applied ammonium (NH4Cl and nitrate (NaNO3 at monthly intervals over a period of 4 years. We collected soil samples from the ammonium and nitrate treatments and control plots in three different seasons, namely spring, summer, and fall, to evaluate the how the biomass of different soil microbial communities and enzyme activities responded to the ammonium (NH4Cl and nitrate (NaNO3 applications. Our results showed that the total contents of phospholipid fatty acids (PLFAs decreased by 24 and 11 % in the ammonium and nitrate treatments, respectively. The inhibitory effects of ammonium on Gram-positive bacteria (G+ and bacteria, fungi, actinomycetes, and arbuscular mycorrhizal fungi (AMF PLFA contents ranged from 14 to 40 % across the three seasons. We also observed that the absolute activities of C, N, and P hydrolyses and oxidases were inhibited by ammonium and nitrate, but that nitrate had stronger inhibitory effects on the activities of acid phosphatase (AP than ammonium. The activities of N-acquisition specific enzymes (enzyme activities normalized by total PLFA contents were about 21 and 43 % lower in the ammonium and nitrate treatments than in the control, respectively. However, the activities of P-acquisition specific enzymes were about 19 % higher in the ammonium treatment than in the control. Using redundancy analysis (RDA, we found that the measured C, N, and P hydrolysis and polyphenol oxidase (PPO activities were positively correlated with the soil pH and ammonium contents, but were negatively correlated with the nitrate contents. The PLFA biomarker contents were positively

  4. Contrasting effects of ammonium and nitrate additions on the biomass of soil microbial communities and enzyme activities in subtropical China

    Science.gov (United States)

    Zhang, Chuang; Zhang, Xin-Yu; Zou, Hong-Tao; Kou, Liang; Yang, Yang; Wen, Xue-Fa; Li, Sheng-Gong; Wang, Hui-Min; Sun, Xiao-Min

    2017-10-01

    The nitrate to ammonium ratios in nitrogen (N) compounds in wet atmospheric deposits have increased over the recent past, which is a cause for some concern as the individual effects of nitrate and ammonium deposition on the biomass of different soil microbial communities and enzyme activities are still poorly defined. We established a field experiment and applied ammonium (NH4Cl) and nitrate (NaNO3) at monthly intervals over a period of 4 years. We collected soil samples from the ammonium and nitrate treatments and control plots in three different seasons, namely spring, summer, and fall, to evaluate the how the biomass of different soil microbial communities and enzyme activities responded to the ammonium (NH4Cl) and nitrate (NaNO3) applications. Our results showed that the total contents of phospholipid fatty acids (PLFAs) decreased by 24 and 11 % in the ammonium and nitrate treatments, respectively. The inhibitory effects of ammonium on Gram-positive bacteria (G+) and bacteria, fungi, actinomycetes, and arbuscular mycorrhizal fungi (AMF) PLFA contents ranged from 14 to 40 % across the three seasons. We also observed that the absolute activities of C, N, and P hydrolyses and oxidases were inhibited by ammonium and nitrate, but that nitrate had stronger inhibitory effects on the activities of acid phosphatase (AP) than ammonium. The activities of N-acquisition specific enzymes (enzyme activities normalized by total PLFA contents) were about 21 and 43 % lower in the ammonium and nitrate treatments than in the control, respectively. However, the activities of P-acquisition specific enzymes were about 19 % higher in the ammonium treatment than in the control. Using redundancy analysis (RDA), we found that the measured C, N, and P hydrolysis and polyphenol oxidase (PPO) activities were positively correlated with the soil pH and ammonium contents, but were negatively correlated with the nitrate contents. The PLFA biomarker contents were positively correlated with soil

  5. 11 Soil Microbial Biomass

    African Journals Online (AJOL)

    186–198. Insam H. (1990). Are the soil microbial biomass and basal respiration governed by the climatic regime? Soil. Biol. Biochem. 22: 525–532. Insam H. D. and Domsch K. H. (1989). Influence of microclimate on soil microbial biomass. Soil Biol. Biochem. 21: 211–21. Jenkinson D. S. (1988). Determination of microbial.

  6. [Study on soil enzyme activities and microbial biomass carbon in greenland irrigated with reclaimed water].

    Science.gov (United States)

    Pan, Neng; Hou, Zhen-An; Chen, Wei-Ping; Jiao, Wen-Tao; Peng, Chi; Liu, Wen

    2012-12-01

    The physicochemical properties of soils might be changed under the long-term reclaimed water irrigation. Its effects on soil biological activities have received great attentions. We collected surface soil samples from urban green spaces and suburban farmlands of Beijing. Soil microbial biomass carbon (SMBC), five types of soil enzyme activities (urease, alkaline phosphatase, invertase, dehydrogenase and catalase) and physicochemical indicators in soils were measured subsequently. SMBC and enzyme activities from green land soils irrigated with reclaimed water were higher than that of control treatments using drinking water, but the difference is not significant in farmland. The SMBC increased by 60.1% and 14.2% than those control treatments in 0-20 cm soil layer of green land and farmland, respectively. Compared with their respective controls, the activities of enzymes in 0-20 cm soil layer of green land and farmland were enhanced by an average of 36.7% and 7.4%, respectively. Investigation of SMBC and enzyme activities decreased with increasing of soil depth. Significantly difference was found between 0-10 cm and 10-20 cm soil layer in green land. Soil biological activities were improved with long-term reclaimed water irrigation in Beijing.

  7. Functional Enzyme-Based Approach for Linking Microbial Community Functions with Biogeochemical Process Kinetics

    Energy Technology Data Exchange (ETDEWEB)

    Li, Minjing [School; Qian, Wei-jun [Pacific Northwest National Laboratory, Richland, Washington 99354, United States; Gao, Yuqian [Pacific Northwest National Laboratory, Richland, Washington 99354, United States; Shi, Liang [School; Liu, Chongxuan [Pacific Northwest National Laboratory, Richland, Washington 99354, United States; School

    2017-09-28

    The kinetics of biogeochemical processes in natural and engineered environmental systems are typically described using Monod-type or modified Monod-type models. These models rely on biomass as surrogates for functional enzymes in microbial community that catalyze biogeochemical reactions. A major challenge to apply such models is the difficulty to quantitatively measure functional biomass for constraining and validating the models. On the other hand, omics-based approaches have been increasingly used to characterize microbial community structure, functions, and metabolites. Here we proposed an enzyme-based model that can incorporate omics-data to link microbial community functions with biogeochemical process kinetics. The model treats enzymes as time-variable catalysts for biogeochemical reactions and applies biogeochemical reaction network to incorporate intermediate metabolites. The sequences of genes and proteins from metagenomes, as well as those from the UniProt database, were used for targeted enzyme quantification and to provide insights into the dynamic linkage among functional genes, enzymes, and metabolites that are necessary to be incorporated in the model. The application of the model was demonstrated using denitrification as an example by comparing model-simulated with measured functional enzymes, genes, denitrification substrates and intermediates

  8. Seasonal Variation in Soil Microbial Biomass, Bacterial Community Composition and Extracellular Enzyme Activity in Relation to Soil Respiration in a Northern Great Plains Grassland

    Science.gov (United States)

    Wilton, E.; Flanagan, L. B.

    2014-12-01

    Soil respiration rate is affected by seasonal changes in temperature and moisture, but is this a direct effect on soil metabolism or an indirect effect caused by changes in microbial biomass, bacterial community composition and substrate availability? In order to address this question, we compared continuous measurements of soil and plant CO2 exchange made with an automatic chamber system to analyses conducted on replicate soil samples collected on four dates during June-August. Microbial biomass was estimated from substrate-induced respiration rate, bacterial community composition was determined by 16S rRNA amplicon pyrosequencing, and β-1,4-N-acetylglucosaminidase (NAGase) and phenol oxidase enzyme activities were assayed fluorometrically or by absorbance measurements, respectively. Soil microbial biomass declined from June to August in strong correlation with a progressive decline in soil moisture during this time period. Soil bacterial species richness and alpha diversity showed no significant seasonal change. However, bacterial community composition showed a progressive shift over time as measured by Bray-Curtis dissimilarity. In particular, the change in community composition was associated with increasing relative abundance in the alpha and delta classes, and declining abundance of the beta and gamma classes of the Proteobacteria phylum during June-August. NAGase showed a progressive seasonal decline in potential activity that was correlated with microbial biomass and seasonal changes in soil moisture. In contrast, phenol oxidase showed highest potential activity in mid-July near the time of peak soil respiration and ecosystem photosynthesis, which may represent a time of high input of carbon exudates into the soil from plant roots. This input of exudates may stimulate the activity of phenol oxidase, a lignolytic enzyme involved in the breakdown of soil organic matter. These analyses indicated that seasonal change in soil respiration is a complex

  9. Assessment of microbial activity and biomass in different soils exposed to nicosulfuron

    Directory of Open Access Journals (Sweden)

    Ljiljana Šantrić

    2014-09-01

    Full Text Available The effects of the herbicide nicosulfuron on the abundance of cellulolytic and proteolytic microorganisms, activity of β-glucosidase and protease enzymes, and microbial phosphorus biomass were examined. A laboratory bioassay was set up on two types of agricultural soils differing in physicochemical properties. The following concentrations were tested: 0.3, 0.6, 3.0 and 30.0 mg a.i./kg of soil. Samples were collected 3, 7, 14, 30 and 45 days after treatment with nicosulfuron. The results showed that nicosulfuron significantly reduced the abundance of cellulolytic microorganisms in both soils, as well as microbial biomass phosphorus in sandy loam soil. The herbicide was found to stimulate β-glucosidase and protease activity in both types of soil and microbial biomass phosphorus in loamy soil. Proteolytic microorganisms remained unaffected by nicosulfuron.

  10. Enzymes for improved biomass conversion

    Science.gov (United States)

    Brunecky, Roman; Himmel, Michael E.

    2016-02-02

    Disclosed herein are enzymes and combinations of the enzymes useful for the hydrolysis of cellulose and the conversion of biomass. Methods of degrading cellulose and biomass using enzymes and cocktails of enzymes are also disclosed.

  11. Renewable biofuels bioconversion of lignocellulosic biomass by microbial community

    CERN Document Server

    Rana, Vandana

    2017-01-01

    This book offers a complete introduction for novices to understand key concepts of biocatalysis and how to produce in-house enzymes that can be used for low-cost biofuels production. The authors discuss the challenges involved in the commercialization of the biofuel industry, given the expense of commercial enzymes used for lignocellulose conversion. They describe the limitations in the process, such as complexity of lignocellulose structure, different microbial communities’ actions and interactions for degrading the recalcitrant structure of lignocellulosic materials, hydrolysis mechanism and potential for bio refinery. Readers will gain understanding of the key concepts of microbial catalysis of lignocellulosic biomass, process complexities and selection of microbes for catalysis or genetic engineering to improve the production of bioethanol or biofuel.

  12. Microbial biomass dynamics dominate N cycle responses to warming in a sub-arctic peatland

    Science.gov (United States)

    Weedon, J. T.; Aerts, R.; Kowalchuk, G. K.; van Bodegom, P. M.

    2012-04-01

    The balance of primary production and decomposition in sub-arctic peatlands may shift with climate change. Nitrogen availability will modulate this shift, but little is known about the drivers of soil nitrogen dynamics in these environments, and how they are influenced by rising soil temperatures. We used a long-term open top chamber warming experiment in Abisko, Sweden, to test for the interactive effects of spring warming, summer warming and winter snow addition on soil organic and inorganic nitrogen fluxes, potential activities of carbon and nitrogen cycle enzymes, and the structure of the soil-borne microbial communities. Summer warming increased the flux of soil organic nitrogen over the growing season, while simultaneously causing a seasonal decrease in microbial biomass, suggesting that N flux is driven by large late-season dieback of microbes. This change in N cycle dynamics was not reflected in any of the measured potential enzyme activities. Moreover, the soil microbial community structure was stable across treatments, suggesting non-specific microbial dieback. To further test whether the observed patterns were driven by direct temperature effects or indirect effects (via microbial biomass dynamics), we conducted follow-up controlled experiments in soil mesocosms. Experimental additions of dead microbial cells had stronger effects on N pool sizes and enzyme activities than either plant litter addition or a 5 °C alteration in incubation temperatures. Peat respiration was positively affected by both substrate addition and higher incubation temperatures, but the temperature-only effect was not sufficient to account for the increases in respiration observed in previous field experiments. We conclude that warming effects on peatland N cycling (and to some extent C cycling) are dominated by indirect effects, acting through alterations to the seasonal flux of microbe-derived organic matter. We propose that climate change models of soil carbon and nitrogen

  13. Targeted quantification of functional enzyme dynamics in environmental samples for microbially mediated biogeochemical processes: Targeted quantification of functional enzyme dynamics

    Energy Technology Data Exchange (ETDEWEB)

    Li, Minjing [School of Environmental Studies, China University of Geosciences, Wuhan 430074 People' s Republic of China; Gao, Yuqian [Pacific Northwest National Laboratory, Richland, WA 99354 USA; Qian, Wei-Jun [Pacific Northwest National Laboratory, Richland, WA 99354 USA; Shi, Liang [Pacific Northwest National Laboratory, Richland, WA 99354 USA; Liu, Yuanyuan [Pacific Northwest National Laboratory, Richland, WA 99354 USA; Nelson, William C. [Pacific Northwest National Laboratory, Richland, WA 99354 USA; Nicora, Carrie D. [Pacific Northwest National Laboratory, Richland, WA 99354 USA; Resch, Charles T. [Pacific Northwest National Laboratory, Richland, WA 99354 USA; Thompson, Christopher [Pacific Northwest National Laboratory, Richland, WA 99354 USA; Yan, Sen [School of Environmental Studies, China University of Geosciences, Wuhan 430074 People' s Republic of China; Fredrickson, James K. [Pacific Northwest National Laboratory, Richland, WA 99354 USA; Zachara, John M. [Pacific Northwest National Laboratory, Richland, WA 99354 USA; Liu, Chongxuan [Pacific Northwest National Laboratory, Richland, WA 99354 USA; School of Environmental Science and Engineering, Southern University of Science and Technology, Shenzhen 518055 People' s Republic of China

    2017-07-13

    Microbially mediated biogeochemical processes are catalyzed by enzymes that control the transformation of carbon, nitrogen, and other elements in environment. The dynamic linkage between enzymes and biogeochemical species transformation has, however, rarely been investigated because of the lack of analytical approaches to efficiently and reliably quantify enzymes and their dynamics in soils and sediments. Herein, we developed a signature peptide-based technique for sensitively quantifying dissimilatory and assimilatory enzymes using nitrate-reducing enzymes in a hyporheic zone sediment as an example. Moreover, the measured changes in enzyme concentration were found to correlate with the nitrate reduction rate in a way different from that inferred from biogeochemical models based on biomass or functional genes as surrogates for functional enzymes. This phenomenon has important implications for understanding and modeling the dynamics of microbial community functions and biogeochemical processes in environments. Our results also demonstrate the importance of enzyme quantification for the identification and interrogation of those biogeochemical processes with low metabolite concentrations as a result of faster enzyme-catalyzed consumption of metabolites than their production. The dynamic enzyme behaviors provide a basis for the development of enzyme-based models to describe the relationship between the microbial community and biogeochemical processes.

  14. Respiration, microbial biomass and soil phosphatase activity in two agroecosystems and one forest in Turrialba, Costa Rica

    Directory of Open Access Journals (Sweden)

    Wuellins Durango

    2015-06-01

    Full Text Available In order to evaluate some microbiological and biochemical characteristics, a comparative study was carried out, as related to 3 different land uses in Ultisols located in Grano de Oro, Turrialba, Costa Rica. Three soil management systems were selected (two agroecosystems, coffee and coffee-banana and forest. In each farm, 4 composite soil samples were collected, on which microbial biomass and respiration, and phosphatase enzyme activity analysis were performed. The microbial biomass in forest was statistically higher (423 mg C kg-1 compared to those in agroecosystems coffee and coffee-banana (77 and 111 mg C kg-1 respectively. Microbial respiration did not show differences due to land management (580, 560 and 570 μg CO2 g-1.day-1 in coffee, coffee-banana and forest systems, respectively. It was also determined that the enzyme phosphatase activity in forest soils was statistically higher (4432 μg p-NP g-1.h-1. The data suggest that soil conditions in the forest favor greater microbial activity and phosphatase biomass, as compared to agricultural systems.

  15. Microbial dynamics and enzyme activities in tropical Andosols depending on land use and nutrient inputs

    Science.gov (United States)

    Mganga, Kevin; Razavi, Bahar; Kuzyakov, Yakov

    2015-04-01

    Microbial decomposition of soil organic matter is mediated by enzymes and is a key source of terrestrial CO2 emissions. Microbial and enzyme activities are necessary to understand soil biochemical functioning and identify changes in soil quality. However, little is known about land use and nutrients availability effects on enzyme activities and microbial processes, especially in tropical soils of Africa. This study was conducted to examine how microbial and enzyme activities differ between different land uses and nutrient availability. As Andosols of Mt. Kilimanjaro are limited by nutrient concentrations, we hypothesize that N and P additions will stimulate enzyme activity. N and P were added to soil samples (0-20 cm) representing common land use types in East Africa: (1) savannah, (2) maize fields, (3) lower montane forest, (4) coffee plantation, (5) grasslands and (6) traditional Chagga homegardens. Total CO2 efflux from soil, microbial biomass and activities of β-glucosidase, cellobiohydrolase, chitinase and phosphatase involved in C, N and P cycling, respectively was monitored for 60 days. Total CO2 production, microbial biomass and enzyme activities varied in the order forest soils > grassland soils > arable soils. Increased β-glucosidase and cellobiohydrolase activities after N addition of grassland soils suggest that microorganisms increased N uptake and utilization to produce C-acquiring enzymes. Low N concentration in all soils inhibited chitinase activity. Depending on land use, N and P addition had an inhibitory or neutral effect on phosphatase activity. We attribute this to the high P retention of Andosols and low impact of N and P on the labile P fractions. Enhanced CO2 production after P addition suggests that increased P availability could stimulate soil organic matter biodegradation in Andosols. In conclusion, land use and nutrients influenced soil enzyme activities and microbial dynamics and demonstrated the decline in soil quality after landuse

  16. Strategies for enhancing the effectiveness of metagenomic-based enzyme discovery in lignocellulytic microbial communities

    Energy Technology Data Exchange (ETDEWEB)

    DeAngelis, K.M.; Gladden, J.G.; Allgaier, M.; D' haeseleer, P.; Fortney, J.L.; Reddy, A.; Hugenholtz, P.; Singer, S.W.; Vander Gheynst, J.; Silver, W.L.; Simmons, B.; Hazen, T.C.

    2010-03-01

    Producing cellulosic biofuels from plant material has recently emerged as a key U.S. Department of Energy goal. For this technology to be commercially viable on a large scale, it is critical to make production cost efficient by streamlining both the deconstruction of lignocellulosic biomass and fuel production. Many natural ecosystems efficiently degrade lignocellulosic biomass and harbor enzymes that, when identified, could be used to increase the efficiency of commercial biomass deconstruction. However, ecosystems most likely to yield relevant enzymes, such as tropical rain forest soil in Puerto Rico, are often too complex for enzyme discovery using current metagenomic sequencing technologies. One potential strategy to overcome this problem is to selectively cultivate the microbial communities from these complex ecosystems on biomass under defined conditions, generating less complex biomass-degrading microbial populations. To test this premise, we cultivated microbes from Puerto Rican soil or green waste compost under precisely defined conditions in the presence dried ground switchgrass (Panicum virgatum L.) or lignin, respectively, as the sole carbon source. Phylogenetic profiling of the two feedstock-adapted communities using SSU rRNA gene amplicon pyrosequencing or phylogenetic microarray analysis revealed that the adapted communities were significantly simplified compared to the natural communities from which they were derived. Several members of the lignin-adapted and switchgrass-adapted consortia are related to organisms previously characterized as biomass degraders, while others were from less well-characterized phyla. The decrease in complexity of these communities make them good candidates for metagenomic sequencing and will likely enable the reconstruction of a greater number of full length genes, leading to the discovery of novel lignocellulose-degrading enzymes adapted to feedstocks and conditions of interest.

  17. Long-term effects of aided phytostabilisation of trace elements on microbial biomass and activity, enzyme activities, and composition of microbial community in the Jales contaminated mine spoils

    Energy Technology Data Exchange (ETDEWEB)

    Renella, Giancarlo [Department of Soil Science and Plant Nutrition, University of Florence, Piazzale delle Cascine 28, I-50144 Florence (Italy)], E-mail: giancarlo.renella@unifi.it; Landi, Loretta; Ascher, Judith; Ceccherini, Maria Teresa; Pietramellara, Giacomo; Mench, Michel; Nannipieri, Paolo [Department of Soil Science and Plant Nutrition, University of Florence, Piazzale delle Cascine 28, I-50144 Florence (Italy)

    2008-04-15

    We studied the effectiveness of remediation on microbial endpoints, namely microbial biomass and activity, microbial and plant species richness, of an As-contaminated mine spoil, amended with compost (C) alone and in combination with beringite (B) or zerovalent iron grit (Z), to increase organic matter content and reduce trace elements mobility, and to allow Holcus lanatus and Pinus pinaster growth. Untreated spoil showed the lowest microbial biomass and activity and hydrolase activities, and H. lanatus as sole plant species, whereas the presented aided phytostabilisation option, especially CBZ treatment, significantly increased microbial biomass and activity and allowed colonisation by several plant species, comparable to those of an uncontaminated sandy soil. Microbial species richness was only increased in spoils amended with C alone. No clear correlation occurred between trace element mobility and microbial parameters and plant species richness. Our results indicate that the choice of indicators of soil remediation practices is a bottleneck. - Organo-mineral amendment and revegetation of a gold mine spoil increased microbial activity but did not increase microbial species richness.

  18. Long-term effects of aided phytostabilisation of trace elements on microbial biomass and activity, enzyme activities, and composition of microbial community in the Jales contaminated mine spoils

    International Nuclear Information System (INIS)

    Renella, Giancarlo; Landi, Loretta; Ascher, Judith; Ceccherini, Maria Teresa; Pietramellara, Giacomo; Mench, Michel; Nannipieri, Paolo

    2008-01-01

    We studied the effectiveness of remediation on microbial endpoints, namely microbial biomass and activity, microbial and plant species richness, of an As-contaminated mine spoil, amended with compost (C) alone and in combination with beringite (B) or zerovalent iron grit (Z), to increase organic matter content and reduce trace elements mobility, and to allow Holcus lanatus and Pinus pinaster growth. Untreated spoil showed the lowest microbial biomass and activity and hydrolase activities, and H. lanatus as sole plant species, whereas the presented aided phytostabilisation option, especially CBZ treatment, significantly increased microbial biomass and activity and allowed colonisation by several plant species, comparable to those of an uncontaminated sandy soil. Microbial species richness was only increased in spoils amended with C alone. No clear correlation occurred between trace element mobility and microbial parameters and plant species richness. Our results indicate that the choice of indicators of soil remediation practices is a bottleneck. - Organo-mineral amendment and revegetation of a gold mine spoil increased microbial activity but did not increase microbial species richness

  19. Identification of novel biomass-degrading enzymes from genomic dark matter: Populating genomic sequence space with functional annotation.

    Science.gov (United States)

    Piao, Hailan; Froula, Jeff; Du, Changbin; Kim, Tae-Wan; Hawley, Erik R; Bauer, Stefan; Wang, Zhong; Ivanova, Nathalia; Clark, Douglas S; Klenk, Hans-Peter; Hess, Matthias

    2014-08-01

    Although recent nucleotide sequencing technologies have significantly enhanced our understanding of microbial genomes, the function of ∼35% of genes identified in a genome currently remains unknown. To improve the understanding of microbial genomes and consequently of microbial processes it will be crucial to assign a function to this "genomic dark matter." Due to the urgent need for additional carbohydrate-active enzymes for improved production of transportation fuels from lignocellulosic biomass, we screened the genomes of more than 5,500 microorganisms for hypothetical proteins that are located in the proximity of already known cellulases. We identified, synthesized and expressed a total of 17 putative cellulase genes with insufficient sequence similarity to currently known cellulases to be identified as such using traditional sequence annotation techniques that rely on significant sequence similarity. The recombinant proteins of the newly identified putative cellulases were subjected to enzymatic activity assays to verify their hydrolytic activity towards cellulose and lignocellulosic biomass. Eleven (65%) of the tested enzymes had significant activity towards at least one of the substrates. This high success rate highlights that a gene context-based approach can be used to assign function to genes that are otherwise categorized as "genomic dark matter" and to identify biomass-degrading enzymes that have little sequence similarity to already known cellulases. The ability to assign function to genes that have no related sequence representatives with functional annotation will be important to enhance our understanding of microbial processes and to identify microbial proteins for a wide range of applications. © 2014 Wiley Periodicals, Inc.

  20. Nitrogen amendment of green waste impacts microbial community, enzyme secretion and potential for lignocellulose decomposition

    Energy Technology Data Exchange (ETDEWEB)

    Yu, Chaowei; Harrold, Duff R.; Claypool, Joshua T.; Simmons, Blake A.; Singer, Steven W.; Simmons, Christopher W.; VanderGheynst, Jean S.

    2017-01-01

    Microorganisms involved in biomass deconstruction are an important resource for organic waste recycling and enzymes for lignocellulose bioconversion. The goals of this paper were to examine the impact of nitrogen amendment on microbial community restructuring, secretion of xylanases and endoglucanases, and potential for biomass deconstruction. Communities were cultivated aerobically at 55 °C on green waste (GW) amended with varying levels of NH4Cl. Bacterial and fungal communities were determined using 16S rRNA and ITS region gene sequencing and PICRUSt (Phylogenetic Investigation of Communities by Reconstruction of Unobserved States) was applied to predict relative abundance of genes involved in lignocellulose hydrolysis. Nitrogen amendment significantly increased secretion of xylanases and endoglucanases, and microbial activity; enzyme activities and cumulative respiration were greatest when nitrogen level in GW was between 4.13–4.56 wt% (g/g), but decreased with higher nitrogen levels. The microbial community shifted to one with increasing potential to decompose complex polymers as nitrogen increased with peak potential occurring between 3.79–4.45 wt% (g/g) nitrogen amendment. Finally, the results will aid in informing the management of nitrogen level to foster microbial communities capable of secreting enzymes that hydrolyze recalcitrant polymers in lignocellulose and yield rapid decomposition of green waste.

  1. Mycelial growth interactions and mannan-degrading enzyme ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-05-18

    May 18, 2009 ... enzymes (Frost and Moss, 1987). However, microbial enzymes are more in use due to cheaper substrates and ease of process modification. In microbial enzyme and biomass production, defined mixed culture method in which more than one organism grows simultaneously can result in increased biomass ...

  2. Contrasting effects of biochar versus manure on soil microbial communities and enzyme activities in an Aridisol.

    Science.gov (United States)

    Elzobair, Khalid A; Stromberger, Mary E; Ippolito, James A; Lentz, Rodrick D

    2016-01-01

    Biochar can increase microbial activity, alter microbial community structure, and increase soil fertility in arid and semi-arid soils, but at relatively high rates that may be impractical for large-scale field studies. This contrasts with organic amendments such as manure, which can be abundant and inexpensive if locally available, and thus can be applied to fields at greater rates than biochar. In a field study comparing biochar and manure, a fast pyrolysis hardwood biochar (22.4 Mg ha(-1)), dairy manure (42 Mg ha(-1) dry wt), a combination of biochar and manure at the aforementioned rates, or no amendment (control) was applied to an Aridisol (n=3) in fall 2008. Plots were annually cropped to corn (Zea maize L.). Surface soils (0-30 cm) were sampled directly under corn plants in late June 2009 and early August 2012, and assayed for microbial community fatty acid methyl ester (FAME) profiles and six extracellular enzyme activities involved in soil C, N, and P cycling. Arbuscular mycorrhizal (AM) fungal colonization was assayed in corn roots in 2012. Biochar had no effect on microbial biomass, community structure, extracellular enzyme activities, or AM fungi root colonization of corn. In the short-term, manure amendment increased microbial biomass, altered microbial community structure, and significantly reduced the relative concentration of the AM fungal biomass in soil. Manure also reduced the percent root colonization of corn by AM fungi in the longer-term. Thus, biochar and manure had contrasting short-term effects on soil microbial communities, perhaps because of the relatively low application rate of biochar. Copyright © 2015 Elsevier Ltd. All rights reserved.

  3. Biomass and enzyme activity of two soil transects at King George Island, Maritime Antarctica

    Czech Academy of Sciences Publication Activity Database

    Tscherko, D.; Bölter, M.; Beyer, L.; Chen, J.; Elster, Josef; Kandeler, E.; Kuhn, D.; Blume, H. P.

    2003-01-01

    Roč. 35, č. 1 (2003), s. 34-47 ISSN 1523-0430 R&D Projects: GA ČR GA205/94/0156; GA AV ČR KSK6005114 Institutional research plan: CEZ:AV0Z6005908 Keywords : Maritime Antarctica * microbial soil biomass * enzyme activity Subject RIV: EF - Botanics Impact factor: 0.954, year: 2003

  4. Functional diversity of carbohydrate-active enzymes enabling a bacterium to ferment plant biomass.

    Science.gov (United States)

    Boutard, Magali; Cerisy, Tristan; Nogue, Pierre-Yves; Alberti, Adriana; Weissenbach, Jean; Salanoubat, Marcel; Tolonen, Andrew C

    2014-11-01

    Microbial metabolism of plant polysaccharides is an important part of environmental carbon cycling, human nutrition, and industrial processes based on cellulosic bioconversion. Here we demonstrate a broadly applicable method to analyze how microbes catabolize plant polysaccharides that integrates carbohydrate-active enzyme (CAZyme) assays, RNA sequencing (RNA-seq), and anaerobic growth screening. We apply this method to study how the bacterium Clostridium phytofermentans ferments plant biomass components including glucans, mannans, xylans, galactans, pectins, and arabinans. These polysaccharides are fermented with variable efficiencies, and diauxies prioritize metabolism of preferred substrates. Strand-specific RNA-seq reveals how this bacterium responds to polysaccharides by up-regulating specific groups of CAZymes, transporters, and enzymes to metabolize the constituent sugars. Fifty-six up-regulated CAZymes were purified, and their activities show most polysaccharides are degraded by multiple enzymes, often from the same family, but with divergent rates, specificities, and cellular localizations. CAZymes were then tested in combination to identify synergies between enzymes acting on the same substrate with different catalytic mechanisms. We discuss how these results advance our understanding of how microbes degrade and metabolize plant biomass.

  5. Seasonal variability of microbial biomass phosphorus in urban soils.

    Science.gov (United States)

    Halecki, W; Gąsiorek, M

    2015-01-01

    Urban soils have been formed through human activities. Seasonal evaluation with time-control procedure are essential for plant, and activity of microorganisms. Therefore, these processes are crucial in the urban area due to geochemical changes in the past years. The purpose of this study was to investigate the changes of content of microbial biomass phosphorus (P) in the top layer of soils throughout the season. In this research, the concentration of microbial biomass P ranged from 0.01 to 6.29 mg·kg(-1). We used single-factor repeated-measure analysis of variance to test the effect of season on microbial biomass P content of selected urban soils. We found no statistically significant differences between the concentration of microbial biomass P in the investigated urban and sub-urban soils during the growing season. This analysis explicitly recognised that environmental urban conditions are steady. Specifically, we have studied how vegetation seasonality and ability of microbial biomass P are useful for detecting quality deviations, which affect the equilibrium of urban soil. In conclusion, seasonal variability of the stringency of assurance across the different compounds of soil reveals, as expected, the stable condition of the urban soils. Seasonal responses in microbial biomass P under urban soil use should establish a framework as a reference to the activity of the microorganisms. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Finding Biomass Degrading Enzymes Through an Activity-Correlated Quantitative Proteomics Platform (ACPP)

    Science.gov (United States)

    Ma, Hongyan; Delafield, Daniel G.; Wang, Zhe; You, Jianlan; Wu, Si

    2017-04-01

    The microbial secretome, known as a pool of biomass (i.e., plant-based materials) degrading enzymes, can be utilized to discover industrial enzyme candidates for biofuel production. Proteomics approaches have been applied to discover novel enzyme candidates through comparing protein expression profiles with enzyme activity of the whole secretome under different growth conditions. However, the activity measurement of each enzyme candidate is needed for confident "active" enzyme assignments, which remains to be elucidated. To address this challenge, we have developed an Activity-Correlated Quantitative Proteomics Platform (ACPP) that systematically correlates protein-level enzymatic activity patterns and protein elution profiles using a label-free quantitative proteomics approach. The ACPP optimized a high performance anion exchange separation for efficiently fractionating complex protein samples while preserving enzymatic activities. The detected enzymatic activity patterns in sequential fractions using microplate-based assays were cross-correlated with protein elution profiles using a customized pattern-matching algorithm with a correlation R-score. The ACPP has been successfully applied to the identification of two types of "active" biomass-degrading enzymes (i.e., starch hydrolysis enzymes and cellulose hydrolysis enzymes) from Aspergillus niger secretome in a multiplexed fashion. By determining protein elution profiles of 156 proteins in A. niger secretome, we confidently identified the 1,4-α-glucosidase as the major "active" starch hydrolysis enzyme (R = 0.96) and the endoglucanase as the major "active" cellulose hydrolysis enzyme (R = 0.97). The results demonstrated that the ACPP facilitated the discovery of bioactive enzymes from complex protein samples in a high-throughput, multiplexing, and untargeted fashion.

  7. A meta-analysis of soil microbial biomass responses to forest disturbances

    Directory of Open Access Journals (Sweden)

    Sandra Robin Holden

    2013-06-01

    Full Text Available Climate warming is likely to increase the frequency and severity of forest disturbances, with uncertain consequences for soil microbial communities and their contribution to ecosystem C dynamics. To address this uncertainty, we conducted a meta-analysis of 139 published soil microbial responses to forest disturbances. These disturbances included abiotic (fire, harvesting, storm and biotic (insect, pathogen disturbances. We hypothesized that soil microbial biomass would decline following forest disturbances, but that abiotic disturbances would elicit greater reductions in microbial biomass than biotic disturbances. In support of this hypothesis, across all published studies, disturbances reduced soil microbial biomass by an average of 29.4%. However, microbial responses differed between abiotic and biotic disturbances. Microbial responses were significantly negative following fires, harvest, and storms (48.7%, 19.1%, and 41.7% reductions in microbial biomass, respectively. In contrast, changes in soil microbial biomass following insect infestation and pathogen-induced tree mortality were non-significant, although biotic disturbances were poorly represented in the literature. When measured separately, fungal and bacterial responses to disturbances mirrored the response of the microbial community as a whole. Changes in microbial abundance following disturbance were significantly positively correlated with changes in microbial respiration. We propose that the differential effect of abiotic and biotic disturbances on microbial biomass may be attributable to differences in soil disruption and organic C removal from forests among disturbance types. Altogether, these results suggest that abiotic forest disturbances may significantly decrease soil microbial abundance, with corresponding consequences for microbial respiration. Further studies are needed on the effect of biotic disturbances on forest soil microbial communities and soil C dynamics.

  8. Impact of Wildfire on Microbial Biomass in Critical Zone Observatory

    Science.gov (United States)

    Murphy, M. A.; Fairbanks, D.; Chorover, J.; Gallery, R. E.; Rich, V. I.

    2014-12-01

    The recovery of the critical zone following disturbances such as wildfire is not fully understood. Wildfires have increased in size and intensity in western US forests in recent years and these fires influence soil microbial communities, both in composition and overall biomass. Studies have typically shown a 50% post-fire decline in overall microbial biomass (µg per g soil) that can persist for years. There is however, some variability in the severity of biomass decline, and its relationship with burn severity and landscape position have not yet been studied. Since microbial biomass has a cascade of impacts in soil systems, from helping control the rate and diversity the biogeochemical processes occurring, to promoting soil fertility, to impacting the nature and structure of soil carbon (C), fire's lasting impact on it is one mechanistic determinant of the overall post-fire recovery of impacted ecosystems. Additionally, microbial biomass measurements hold potential for testing and incorporation into land surface models (NoahMP, CLM, etc.) in order to improve estimates of long-term effects of climate change and disturbances such as fire on the C cycle. In order to refine our understanding of the impact of fire on microbial biomass and then relate that to biogeochemical processes and ecosystem recovery, we used chloroform fumigation extraction to quantify total microbial biomass C (Cmic ). One year after the June 2013 Thompson Ridge fire in the Jemez River Basin Critical Zone Observatory, we are measuring the Cmic of 22 sites across a gradient of burn severities and 4 control unburned sites, from six depth intervals at each site (0-2, 2-5, 5-10, 10-20, 20-30, and 30-40 cm). We hypothesize that the decrease in microbial biomass in burned sites relative to control sites will correlate with changes in soil biogeochemistry related to burn severity; and that the extent of the impact on biomass will be inversely related to depth in the soil column. Additionally, as the

  9. Role of Bioreactors in Microbial Biomass and Energy Conversion

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Liang [Chongqing University, Chongqing, China; Zhang, Biao [Chongqing University, Chongqing, China; Zhu, Xun [Chongqing University, Chongqing, China; Chang, Haixing [Chongqing University of Technology; Ou, Shawn [ORNL; Wang, HONG [Chongqing University, Chongqing, China

    2018-04-01

    Bioenergy is the world’s largest contributor to the renewable and sustainable energy sector, and it plays a significant role in various energy industries. A large amount of research has contributed to the rapidly evolving field of bioenergy and one of the most important topics is the use of the bioreactor. Bioreactors play a critical role in the successful development of technologies for microbial biomass cultivation and energy conversion. In this chapter, after a brief introduction to bioreactors (basic concepts, configurations, functions, and influencing factors), the applications of the bioreactor in microbial biomass, microbial biofuel conversion, and microbial electrochemical systems are described. Importantly, the role and significance of the bioreactor in the bioenergy process are discussed to provide a better understanding of the use of bioreactors in managing microbial biomass and energy conversion.

  10. Soil Rhizosphere Microbial Communities and Enzyme Activities under Organic Farming in Alabama

    Directory of Open Access Journals (Sweden)

    Zachary Senwo

    2011-07-01

    Full Text Available Evaluation of the soil rhizosphere has been limited by the lack of robust assessments that can explore the vast complex structure and diversity of soil microbial communities. Our objective was to combine fatty acid methyl ester (FAME and pyrosequencing techniques to evaluate soil microbial community structure and diversity. In addition, we evaluated biogeochemical functionality of the microbial communities via enzymatic activities of nutrient cycling. Samples were taken from a silt loam at 0–10 and 10–20 cm in an organic farm under lettuce (Lactuca sativa, potato (Solanum tuberosum, onion (Allium cepa L, broccoli (Brassica oleracea var. botrytis and Tall fescue pasture grass (Festuca arundinacea. Several FAMEs (a15:0, i15:0, i15:1, i16:0, a17:0, i17:0, 10Me17:0, cy17:0, 16:1ω5c and 18:1ω9c varied among the crop rhizospheres. FAME profiles of the soil microbial community under pasture showed a higher fungal:bacterial ratio compared to the soil under lettuce, potato, onion, and broccoli. Soil under potato showed higher sum of fungal FAME indicators compared to broccoli, onion and lettuce. Microbial biomass C and enzyme activities associated with pasture and potato were higher than the other rhizospheres. The lowest soil microbial biomass C and enzyme activities were found under onion. Pyrosequencing revealed significant differences regarding the maximum operational taxonomic units (OTU at 3% dissimilarity level (roughly corresponding to the bacterial species level at 0–10 cm (581.7–770.0 compared to 10–20 cm (563.3–727.7 soil depths. The lowest OTUs detected at 0–10 cm were under broccoli (581.7; whereas the lowest OTUs found at 10–20 cm were under potato (563.3. The predominant phyla (85% in this soil at both depths were Bacteroidetes (i.e., Flavobacteria, Sphingobacteria, and Proteobacteria. Flavobacteriaceae and Xanthomonadaceae were predominant under broccoli. Rhizobiaceae, Hyphomicrobiaceae, and Acidobacteriaceae were more

  11. Microbial respiration per unit microbial biomass increases with carbon-to-nutrient ratios in soils

    Science.gov (United States)

    Spohn, Marie; Chodak, Marcin

    2015-04-01

    The ratio of carbon-to-nutrient in forest floors is usually much higher than the ratio of carbon-to-nutrient that soil microorganisms require for their nutrition. In order to understand how this mismatch affects carbon cycling, the respiration rate per unit soil microbial biomass carbon - the metabolic quotient (qCO2) - was studied. This was done in a field study (Spohn and Chodak, 2015) and in a meta-analysis of published data (Spohn, 2014). Cores of beech, spruce, and mixed spruce-beech forest soils were cut into slices of 1 cm from the top of the litter layer down to 5 cm in the mineral soil, and the relationship between the qCO2 and the soil carbon-to-nitrogen (C:N) and the soil carbon-to-phosphorus (C:P) ratio was analyzed. We found that the qCO2 was positively correlated with soil C:N ratio in spruce soils (R = 0.72), and with the soil C:P ratio in beech (R = 0.93), spruce (R = 0.80) and mixed forest soils (R = 0.96). We also observed a close correlation between the qCO2 and the soil C concentration in all three forest types. Yet, the qCO2 decreased less with depth than the C concentration in all three forest types, suggesting that the change in qCO2 is not only controlled by the soil C concentration. We conclude that microorganisms increase their respiration rate per unit biomass with increasing soil C:P ratio and C concentration, which adjusts the substrate to their nutritional demands in terms of stoichiometry. In an analysis of literature data, I tested the effect of the C:N ratio of soil litter layers on microbial respiration in absolute terms and per unit microbial biomass C. For this purpose, a global dataset on the microbial respiration rate per unit microbial biomass C - termed the metabolic quotient (qCO2) - was compiled form literature data. It was found that the qCO2 in the soil litter layers was positively correlated with the litter C:N ratio and negatively related with the litter nitrogen (N) concentration. The positive relation between the qCO2

  12. Extraction of solubles from plant biomass for use as microbial growth stimulant and methods related thereto

    Energy Technology Data Exchange (ETDEWEB)

    Lau, Ming Woei

    2015-12-08

    A method for producing a microbial growth stimulant (MGS) from a plant biomass is described. In one embodiment, an ammonium hydroxide solution is used to extract a solution of proteins and ammonia from the biomass. Some of the proteins and ammonia are separated from the extracted solution to provide the MGS solution. The removed ammonia can be recycled and the proteins are useful as animal feeds. In one embodiment, the method comprises extracting solubles from pretreated lignocellulosic biomass with a cellulase enzyme-producing growth medium (such T. reesei) in the presence of water and an aqueous extract.

  13. Measurement and characteristics of microbial biomass in forest soils

    International Nuclear Information System (INIS)

    Vance, E.D.

    1986-01-01

    The soil microbial biomass is the primary agent responsible for the breakdown and mineralization of soil organic matter and plays a major role in regulating nutrient availability to plants. In this study, methods for measuring biomass in soil were compared and tested in forest soils ranging in pH from 3.2 to 7.2. A good relationship between biomass C measured using the chloroform fumigation-incubation method and soil ATP or microbial biomass C by direct microscopy was found in soils at or above pH 4.2. The fumigation-incubation method consistently underestimated biomass C in soils below pH 4.2, however. Hypotheses for the breakdown of the fumigation-incubation method in strongly acid soils were tested by using an alterative fumigant, measuring the proportion of added 14 C labelled fungi and bacteria decomposed in fumigated soils (k/sub C/), and by studying the effect of large, non-fumigated soil inocula on the flush of respiration following fumigation. These studies indicated that the failure of the method in strongly acid soils was due to inhibited decomposition of non-microbial soil organic matter by the microbial recolonizing population following fumigation. A modified method for measuring biomass C by fumigation-incubation in acid soils is proposed

  14. High Potential Source for Biomass Degradation Enzyme Discovery and Environmental Aspects Revealed through Metagenomics of Indian Buffalo Rumen

    Directory of Open Access Journals (Sweden)

    K. M. Singh

    2014-01-01

    Full Text Available The complex microbiomes of the rumen functions as an effective system for plant cell wall degradation, and biomass utilization provide genetic resource for degrading microbial enzymes that could be used in the production of biofuel. Therefore the buffalo rumen microbiota was surveyed using shot gun sequencing. This metagenomic sequencing generated 3.9 GB of sequences and data were assembled into 137270 contiguous sequences (contigs. We identified potential 2614 contigs encoding biomass degrading enzymes including glycoside hydrolases (GH: 1943 contigs, carbohydrate binding module (CBM: 23 contigs, glycosyl transferase (GT: 373 contigs, carbohydrate esterases (CE: 259 contigs, and polysaccharide lyases (PE: 16 contigs. The hierarchical clustering of buffalo metagenomes demonstrated the similarities and dissimilarity in microbial community structures and functional capacity. This demonstrates that buffalo rumen microbiome was considerably enriched in functional genes involved in polysaccharide degradation with great prospects to obtain new molecules that may be applied in the biofuel industry.

  15. Where do the immunostimulatory effects of oral proteolytic enzymes ('systemic enzyme therapy') come from? Microbial proteolysis as a possible starting point.

    Science.gov (United States)

    Biziulevicius, Gediminas A

    2006-01-01

    Enteric-coated proteolytic enzyme preparations like Wobenzym and Phlogenzym are widely used for the so-called 'systemic enzyme therapy' both in humans and animals. Numerous publications reveal that oral proteolytic enzymes are able to stimulate directly the activity of immune competent cells as well as to increase efficiency of some of their products. But origins of the immunostimulatory effects of oral proteolytic enzymes are still unclear. The hypothesis described here suggests that it may be proteolysis of intestinal microorganisms that makes the immune competent cells to work in the immunostimulatory manner. The hypothesis was largely formed by several scientific observations: First, microbial lysis products (lipopolysaccharides, muropeptides and other peptidoglycan fragments, beta-glucans, etc.) are well known for their immunostimulatory action. Second, a normal human being hosts a mass of intestinal microorganisms equivalent to about 1 kg. The biomass (mainly due to naturally occurring autolysis) continuously supplies the host's organism with immunostimulatory microbial cell components. Third, the immunostimulatory effects resulting from the oral application of exogenously acting antimicrobial (lytic) enzyme preparations, such as lysozyme and lysosubtilin, are likely to be a result of the action of microbial lysis products. Fourth, cell walls of most microorganisms contain a considerable amount of proteins/peptides, a possible target for exogenous proteolytic enzymes. In fact, several authors have already shown that a number of proteases possess an ability to lyse the microbial cells in vitro. Fifth, the pretreatment of microbial cells (at least of some species) in vitro with proteolytic enzymes makes them more sensitive to the lytic action of lysozyme and, otherwise, pretreatment with lysozyme makes them more susceptible to proteolytic degradation. Sixth, exogenous proteases, when in the intestines, may participate in final steps of food-protein digestion

  16. Elevated atmospheric CO2 increases microbial growth rates and enzymes activity in soil

    Science.gov (United States)

    Blagodatskaya, Evgenia; Blagodatsky, Sergey; Dorodnikov, Maxim; Kuzyakov, Yakov

    2010-05-01

    Increasing the belowground translocation of assimilated carbon by plants grown under elevated CO2 can cause a shift in the structure and activity of the microbial community responsible for the turnover of organic matter in soil. We investigated the long-term effect of elevated CO2 in the atmosphere on microbial biomass and specific growth rates in root-free and rhizosphere soil. The experiments were conducted under two free air carbon dioxide enrichment (FACE) systems: in Hohenheim and Braunschweig, as well as in the intensively managed forest mesocosm of the Biosphere 2 Laboratory (B2L) in Oracle, AZ. Specific microbial growth rates (μ) were determined using the substrate-induced respiration response after glucose and/or yeast extract addition to the soil. We evaluated the effect of elevated CO2 on b-glucosidase, chitinase, phosphatase, and sulfatase to estimate the potential enzyme activity after soil amendment with glucose and nutrients. For B2L and both FACE systems, up to 58% higher μ were observed under elevated vs. ambient CO2, depending on site, plant species and N fertilization. The μ-values increased linearly with atmospheric CO2 concentration at all three sites. The effect of elevated CO2 on rhizosphere microorganisms was plant dependent and increased for: Brassica napus=Triticum aestivumyeast extract then for those growing on glucose, i.e. the effect of elevated CO2 was smoothed on rich vs. simple substrate. So, the r/K strategies ratio can be better revealed by studying growth on simple (glucose) than on rich substrate mixtures (yeast extract). After adding glucose, enzyme activities under elevated CO2 were 1.2-1.9-fold higher than under ambient CO2. This indicates the increased activity of microorganisms, which leads to accelerated C turnover in soil under elevated CO2. Our results clearly showed that the functional characteristics of the soil microbial community (i.e. specific growth rates and enzymes activity) rather than total microbial biomass

  17. Measures of Microbial Biomass for Soil Carbon Decomposition Models

    Science.gov (United States)

    Mayes, M. A.; Dabbs, J.; Steinweg, J. M.; Schadt, C. W.; Kluber, L. A.; Wang, G.; Jagadamma, S.

    2014-12-01

    Explicit parameterization of the decomposition of plant inputs and soil organic matter by microbes is becoming more widely accepted in models of various complexity, ranging from detailed process models to global-scale earth system models. While there are multiple ways to measure microbial biomass, chloroform fumigation-extraction (CFE) is commonly used to parameterize models.. However CFE is labor- and time-intensive, requires toxic chemicals, and it provides no specific information about the composition or function of the microbial community. We investigated correlations between measures of: CFE; DNA extraction yield; QPCR base-gene copy numbers for Bacteria, Fungi and Archaea; phospholipid fatty acid analysis; and direct cell counts to determine the potential for use as proxies for microbial biomass. As our ultimate goal is to develop a reliable, more informative, and faster methods to predict microbial biomass for use in models, we also examined basic soil physiochemical characteristics including texture, organic matter content, pH, etc. to identify multi-factor predictive correlations with one or more measures of the microbial community. Our work will have application to both microbial ecology studies and the next generation of process and earth system models.

  18. BIOMASS AND MICROBIAL ACTIVITY UNDER DIFFERENT FOREST COVERS

    Directory of Open Access Journals (Sweden)

    Rafael Malfitano Braga

    2016-06-01

    Full Text Available This study evaluated the soil fertility, biomass and microbial activity of the soil under forest cover of Eucalyptus grandis, Eucalyptus pilularis, Eucalyptus cloeziana and Corymbia maculata; Pinus Caribbean var. hondurensis, 40 years old, and a fragment of Semideciduous Forest, located on the campus of the Federal University of Lavras. In soil samples collected in the 0-5 cm layer were determined fertility parameters, basal respiration and microbial biomass carbon. The results showed that for the species E. grandis and E. cloeziana the carbon of biomass microbial content was higher than for any other ecosystem evaluated, and equal to those observed under native forest. In contrast, the ground under Pinus had the lowest microbiological indexes. Under C. maculata and E. pilularis the contents were intermediate for this parameter. The basal respiration of all ecosystems was equal. The fertility level was very low in all types of evaluated vegetation.

  19. Plant carbohydrate binding module enhances activity of hybrid microbial cellulase enzyme

    Directory of Open Access Journals (Sweden)

    Caitlin Siobhan Byrt

    2012-11-01

    Full Text Available A synthetic, highly active cellulase enzyme suitable for in planta production may be a valuable tool for biotechnological approaches to develop transgenic biofuel crops with improved digestibility. Here, we demonstrate that the addition of a plant derived carbohydrate binding module (CBM to a synthetic glycosyl hydrolase (GH improved the activity of the hydrolase in releasing sugar from plant biomass. A CEL-HYB1-CBM enzyme was generated by fusing a hybrid microbial cellulase, CEL-HYB1, with the carbohydrate-binding module (CBM of the tomato (Solanum lycopersicum SlCel9C1 cellulase. CEL-HYB1 and CEL-HYB1-CBM enzymes were produced in vitro using Pichia pastoris and the activity of these enzymes was tested using CMC, MUC and native crystalline cellulose assays. The presence of the CBM substantially improved the endo-glucanase activity of CEL-HYB1, especially against the native crystalline cellulose encountered in Sorghum plant cell walls. These results indicate that addition of an endogenous plant derived CBM to cellulase enzymes may enhance hydrolytic activity.

  20. Production by Tobacco Transplastomic Plants of Recombinant Fungal and Bacterial Cell-Wall Degrading Enzymes to Be Used for Cellulosic Biomass Saccharification.

    Science.gov (United States)

    Longoni, Paolo; Leelavathi, Sadhu; Doria, Enrico; Reddy, Vanga Siva; Cella, Rino

    2015-01-01

    Biofuels from renewable plant biomass are gaining momentum due to climate change related to atmospheric CO2 increase. However, the production cost of enzymes required for cellulosic biomass saccharification is a major limiting step in this process. Low-cost production of large amounts of recombinant enzymes by transgenic plants was proposed as an alternative to the conventional microbial based fermentation. A number of studies have shown that chloroplast-based gene expression offers several advantages over nuclear transformation due to efficient transcription and translation systems and high copy number of the transgene. In this study, we expressed in tobacco chloroplasts microbial genes encoding five cellulases and a polygalacturonase. Leaf extracts containing the recombinant enzymes showed the ability to degrade various cell-wall components under different conditions, singly and in combinations. In addition, our group also tested a previously described thermostable xylanase in combination with a cellulase and a polygalacturonase to study the cumulative effect on the depolymerization of a complex plant substrate. Our results demonstrate the feasibility of using transplastomic tobacco leaf extracts to convert cell-wall polysaccharides into reducing sugars, fulfilling a major prerequisite of large scale availability of a variety of cell-wall degrading enzymes for biofuel industry.

  1. Discovery of enzymes for toluene synthesis from anoxic microbial communities

    DEFF Research Database (Denmark)

    Beller, Harry R.; Rodrigues, Andria V.; Zargar, Kamrun

    2018-01-01

    Microbial toluene biosynthesis was reported in anoxic lake sediments more than three decades ago, but the enzyme catalyzing this biochemically challenging reaction has never been identified. Here we report the toluene-producing enzyme PhdB, a glycyl radical enzyme of bacterial origin that catalyzes...... phenylacetate decarboxylation, and its cognate activating enzyme PhdA, a radical S-adenosylmethionine enzyme, discovered in two distinct anoxic microbial communities that produce toluene. The unconventional process of enzyme discovery from a complex microbial community (>300,000 genes), rather than from...... a microbial isolate, involved metagenomics- and metaproteomics-enabled biochemistry, as well as in vitro confirmation of activity with recombinant enzymes. This work expands the known catalytic range of glycyl radical enzymes (only seven reaction types had been characterized previously) and aromatic...

  2. Quantitative analysis of microbial biomass yield in aerobic bioreactor.

    Science.gov (United States)

    Watanabe, Osamu; Isoda, Satoru

    2013-12-01

    We have studied the integrated model of reaction rate equations with thermal energy balance in aerobic bioreactor for food waste decomposition and showed that the integrated model has the capability both of monitoring microbial activity in real time and of analyzing biodegradation kinetics and thermal-hydrodynamic properties. On the other hand, concerning microbial metabolism, it was known that balancing catabolic reactions with anabolic reactions in terms of energy and electron flow provides stoichiometric metabolic reactions and enables the estimation of microbial biomass yield (stoichiometric reaction model). We have studied a method for estimating real-time microbial biomass yield in the bioreactor during food waste decomposition by combining the integrated model with the stoichiometric reaction model. As a result, it was found that the time course of microbial biomass yield in the bioreactor during decomposition can be evaluated using the operational data of the bioreactor (weight of input food waste and bed temperature) by the combined model. The combined model can be applied to manage a food waste decomposition not only for controlling system operation to keep microbial activity stable, but also for producing value-added products such as compost on optimum condition. Copyright © 2013 The Research Centre for Eco-Environmental Sciences, Chinese Academy of Sciences. Published by Elsevier B.V. All rights reserved.

  3. Development of a commercial enzymes system for lignocellulosic biomass saccharification

    Energy Technology Data Exchange (ETDEWEB)

    Kumar, Manoj

    2012-12-20

    DSM Innovation Inc., in its four year effort was able to evaluate and develop its in-house DSM fungal cellulolytic enzymes system to reach enzyme efficiency mandates set by DoE Biomass program MYPP goals. DSM enzyme cocktail is uniquely active at high temperature and acidic pH, offering many benefits and product differentiation in 2G bioethanol production. Under this project, strain and process development, ratio optimization of enzymes, protein and genetic engineering has led to multitudes of improvement in productivity and efficiency making development of a commercial enzyme system for lignocellulosic biomass saccharification viable. DSM is continuing further improvement by additional biodiversity screening, protein engineering and overexpression of enzymes to continue to further lower the cost of enzymes for saccharification of biomass.

  4. Microbial respiration and kinetics of extracellular enzymes activities through rhizosphere and detritusphere at agricultural site

    Science.gov (United States)

    Löppmann, Sebastian; Blagodatskaya, Evgenia; Kuzyakov, Yakov

    2014-05-01

    Rhizosphere and detritusphere are soil microsites with very high resource availability for microorganisms affecting their biomass, composition and functions. In the rhizosphere low molecular compounds occur with root exudates and low available polymeric compounds, as belowground plant senescence. In detritusphere the substrate for decomposition is mainly a polymeric material of low availability. We hypothesized that microorganisms adapted to contrasting quality and availability of substrates in the rhizosphere and detritusphere are strongly different in affinity of hydrolytic enzymes responsible for decomposition of organic compounds. According to common ecological principles easily available substrates are quickly consumed by microorganisms with enzymes of low substrate affinity (i.e. r-strategists). The slow-growing K-strategists with enzymes of high substrate affinity are better adapted for growth on substrates of low availability. Estimation of affinity of enzyme systems to the substrate is based on Michaelis-Menten kinetics, reflecting the dependency of decomposition rates on substrate amount. As enzymes-mediated reactions are substrate-dependent, we further hypothesized that the largest differences in hydrolytic activity between the rhizosphere and detritusphere occur at substrate saturation and that these differences are smoothed with increasing limitation of substrate. Affected by substrate limitation, microbial species follow a certain adaptation strategy. To achieve different depth gradients of substrate availability 12 plots on an agricultural field were established in the north-west of Göttingen, Germany: 1) 4 plots planted with maize, reflecting lower substrate availability with depth; 2) 4 unplanted plots with maize litter input (0.8 kg m-2 dry maize residues), corresponding to detritusphere; 3) 4 bare fallow plots as control. Maize litter was grubbed homogenously into the soil at the first 5 cm to ensure comparable conditions for the herbivore and

  5. Applications of Microbial Enzymes in Food Industry

    Directory of Open Access Journals (Sweden)

    Binod Parameswaran

    2018-01-01

    Full Text Available The use of enzymes or microorganisms in food preparations is an age-old process. With the advancement of technology, novel enzymes with wide range of applications and specificity have been developed and new application areas are still being explored. Microorganisms such as bacteria, yeast and fungi and their enzymes are widely used in several food preparations for improving the taste and texture and they offer huge economic benefits to industries. Microbial enzymes are the preferred source to plants or animals due to several advantages such as easy, cost-effective and consistent production. The present review discusses the recent advancement in enzyme technology for food industries. A comprehensive list of enzymes used in food processing, the microbial source of these enzymes and the wide range of their application are discussed.

  6. [Effects of different application rates of calcium cyanamide on soil microbial biomass and enzyme activity in cucumber continuous cropping].

    Science.gov (United States)

    Zhang, Xue-peng; Ning, Tang-yuan; Yang, Yan; Sun, Tao; Zhang, Shu-min; Wang, Bin

    2015-10-01

    A 2-year field experiment was conducted to study the effects of CaCN2 combined with cucumber straw retention on soil microbial biomass carbon (SMBC) , soil microbial biomass nitrogen (SMBN) and soil enzyme activities under cucumber continuous cropping system. Four treatments were used in this study as follows: CK (null CaCN2), CaCN2-90 (1350 kg CaCN2 . hm-2) CaCN2-60 (900 kg CaCN2 . hm-2), CaCN2-30 (450 kg CaCN2 . hm-2). The results indicated that, compared with the other treatments, CaCN2-90 treatment significantly decreased SMBC in 0-10 cm soil layer at seedling stage, but increased SMBC in 0-20 cm soil layer after early-fruit stage. Compared with CK, CaCN2 increased SMBC in 0-20 cm soil layer at late-fruit stage, and increased SMBN in 0-10 cm soil layer at mid- and late-fruit stages, however there was no significant trend among CaCN2 treatments in the first year (2012), while in the second year (2013) SMBN increased with the increasing CaCN2 amount after mid-fruit stage. CaCN2 increased straw decaying and nutrients releasing, and also increased soil organic matter. Furthermore, the CaCN2-90 could accelerate straw decomposition. Compared with CK, CaCN2 effectively increased soil urease, catalase and polyphenol oxidase activity. The soil urease activity increased while the polyphenol oxidase activity decreased with the increase of CaCN2, and CaCN2-60 could significantly improve catalase activity. Soil organic matter, urease activity and catalase activity had significant positive correlations with SMBC and SMBN. However, polyphenol oxidase activity was negatively correlated to SMBC and SMBN. Our findings indicated that CaCN2 application at 900 kg . hm-2 combined with cucumber straw retention could effectively improve soil environment, alleviating the soil obstacles under the cucumber continuous cropping system.

  7. A global analysis of soil microbial biomass carbon, nitrogen and phosphorus in terrestrial ecosystems

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Xiaofeng [ORNL; Thornton, Peter E [ORNL; Post, Wilfred M [ORNL

    2013-01-01

    Soil microbes play a pivotal role in regulating land-atmosphere interactions; the soil microbial biomass carbon (C), nitrogen (N), phosphorus (P) and C:N:P stoichiometry are important regulators for soil biogeochemical processes; however, the current knowledge on magnitude, stoichiometry, storage, and spatial distribution of global soil microbial biomass C, N, and P is limited. In this study, 3087 pairs of data points were retrieved from 281 published papers and further used to summarize the magnitudes and stoichiometries of C, N, and P in soils and soil microbial biomass at global- and biome-levels. Finally, global stock and spatial distribution of microbial biomass C and N in 0-30 cm and 0-100 cm soil profiles were estimated. The results show that C, N, and P in soils and soil microbial biomass vary substantially across biomes; the fractions of soil nutrient C, N, and P in soil microbial biomass are 1.6% in a 95% confidence interval of (1.5%-1.6%), 2.9% in a 95% confidence interval of (2.8%-3.0%), and 4.4% in a 95% confidence interval of (3.9%-5.0%), respectively. The best estimates of C:N:P stoichiometries for soil nutrients and soil microbial biomass are 153:11:1, and 47:6:1, respectively, at global scale, and they vary in a wide range among biomes. Vertical distribution of soil microbial biomass follows the distribution of roots up to 1 m depth. The global stock of soil microbial biomass C and N were estimated to be 15.2 Pg C and 2.3 Pg N in the 0-30 cm soil profiles, and 21.2 Pg C and 3.2 Pg N in the 0-100 cm soil profiles. We did not estimate P in soil microbial biomass due to data shortage and insignificant correlation with soil total P and climate variables. The spatial patterns of soil microbial biomass C and N were consistent with those of soil organic C and total N, i.e. high density in northern high latitude, and low density in low latitudes and southern hemisphere.

  8. Short-term parasite-infection alters already the biomass, activity and functional diversity of soil microbial communities

    Science.gov (United States)

    Li, Jun-Min; Jin, Ze-Xin; Hagedorn, Frank; Li, Mai-He

    2014-11-01

    Native parasitic plants may be used to infect and control invasive plants. We established microcosms with invasive Mikania micrantha and native Coix lacryma-jobi growing in mixture on native soils, with M. micrantha being infected by parasitic Cuscuta campestris at four intensity levels for seven weeks to estimate the top-down effects of plant parasitism on the biomass and functional diversity of soil microbial communities. Parasitism significantly decreased root biomass and altered soil microbial communities. Soil microbial biomass decreased, but soil respiration increased at the two higher infection levels, indicating a strong stimulation of soil microbial metabolic activity (+180%). Moreover, a Biolog assay showed that the infection resulted in a significant change in the functional diversity indices of soil microbial communities. Pearson correlation analysis indicated that microbial biomass declined significantly with decreasing root biomass, particularly of the invasive M. micrantha. Also, the functional diversity indices of soil microbial communities were positively correlated with soil microbial biomass. Therefore, the negative effects on the biomass, activity and functional diversity of soil microbial community by the seven week long plant parasitism was very likely caused by decreased root biomass and root exudation of the invasive M. micrantha.

  9. Production by Tobacco Transplastomic Plants of Recombinant Fungal and Bacterial Cell-Wall Degrading Enzymes to Be Used for Cellulosic Biomass Saccharification

    Directory of Open Access Journals (Sweden)

    Paolo Longoni

    2015-01-01

    Full Text Available Biofuels from renewable plant biomass are gaining momentum due to climate change related to atmospheric CO2 increase. However, the production cost of enzymes required for cellulosic biomass saccharification is a major limiting step in this process. Low-cost production of large amounts of recombinant enzymes by transgenic plants was proposed as an alternative to the conventional microbial based fermentation. A number of studies have shown that chloroplast-based gene expression offers several advantages over nuclear transformation due to efficient transcription and translation systems and high copy number of the transgene. In this study, we expressed in tobacco chloroplasts microbial genes encoding five cellulases and a polygalacturonase. Leaf extracts containing the recombinant enzymes showed the ability to degrade various cell-wall components under different conditions, singly and in combinations. In addition, our group also tested a previously described thermostable xylanase in combination with a cellulase and a polygalacturonase to study the cumulative effect on the depolymerization of a complex plant substrate. Our results demonstrate the feasibility of using transplastomic tobacco leaf extracts to convert cell-wall polysaccharides into reducing sugars, fulfilling a major prerequisite of large scale availability of a variety of cell-wall degrading enzymes for biofuel industry.

  10. Increased resiliency and activity of microbial mediated carbon cycling enzymes in diversified bioenergy cropping systems

    Science.gov (United States)

    Upton, R.; Bach, E.; Hofmockel, K. S.

    2017-12-01

    Microbes are mediators of soil carbon (C) and are influenced in membership and activity by nitrogen (N) fertilization and inter-annual abiotic factors. Microbial communities and their extracellular enzyme activities (EEA) are important parameters that influence ecosystem C cycling properties and are often included in microbial explicit C cycling models. In an effort to generate model relevant, empirical findings, we investigated how both microbial community structure and C degrading enzyme activity are influenced by inter-annual variability and N inputs in bioenergy crops. Our study was performed at the Comparison of Biofuel Systems field-site from 2011 to 2014, in three bioenergy cropping systems, continuous corn (CC) and two restored prairies, both fertilized (FP) and unfertilized (P). We hypothesized microbial community structure would diverge during the prairie restoration, leading to changes in C cycling enzymes over time. Using a sequencing approach (16S and ITS) we determined the bacterial and fungal community structure response to the cropping system, fertilization, and inter-annual variability. Additionally, we used EEA of β-glucosidase, cellobiohydrolase, and β-xylosidase to determine inter-annual and ecosystem impacts on microbial activity. Our results show cropping system was a main effect for microbial community structure, with corn diverging from both prairies to be less diverse. Inter-annual changes showed that a drought occurring in 2012 significantly impacted microbial community structure in both the P and CC, decreasing microbial richness. However, FP increased in microbial richness, suggesting the application of N increased resiliency to drought. Similarly, the only year in which C cycling enzymes were impacted by ecosystem was 2012, with FP supporting higher potential enzymatic activity then CC and P. The highest EEA across all ecosystems occurred in 2014, suggesting the continued root biomass and litter build-up in this no till system

  11. Strategies for enzyme saving during saccharification of pretreated lignocellulo-starch biomass: effect of enzyme dosage and detoxification chemicals

    Directory of Open Access Journals (Sweden)

    M.G. Mithra

    2017-08-01

    Full Text Available Two strategies leading to enzyme saving during saccharification of pretreated lignocellulo-starch biomass (LCSB was investigated which included reducing enzyme dosage by varying their levels in enzyme cocktails and enhancing the fermentable sugar yield in enzyme-reduced systems using detoxification chemicals. Time course release of reducing sugars (RS during 24–120 h was significantly higher when an enzyme cocktail containing full dose of cellulase (16 FPU/g cellulose along with half dose each of xylanase (1.5 mg protein/g hemicelluloses and Stargen (12.5 μl/g biomass was used to saccharify conventional dilute sulphuric acid (DSA pretreated biomass compared to a parallel system where only one-fourth the dose of the latter two enzymes was used. The reduction in RS content in the 120 h saccharified mash to the extent of 3–4 g/L compared to the system saccharified with full complement of the three enzymes could be overcome considerably by supplementing the system (half dose of two enzymes with detoxification chemical mix incorporating Tween 20, PEG 4000 and sodium borohydride. Microwave (MW-assisted DSA pretreated biomass on saccharification with enzyme cocktail having full dose of cellulase and half dose of Stargen along with detoxification chemicals gave significantly higher RS yield than DSA pretreated system saccharified using three enzymes. The study showed that xylanase could be eliminated during saccharification of MW-assisted DSA pretreated biomass without affecting RS yield when detoxification chemicals were also supplemented. The Saccharification Efficiency and Overall Conversion Efficiency were also high for the MW-assisted DSA pretreated biomass. Since whole slurry saccharifcation of pretreated biomass is essential to conserve fermentable sugars in LCSB saccharification, detoxification of soluble inhibitors is equally important as channelling out of insoluble lignin remaining in the residue. As one of the major factors contributing

  12. Ambient ultraviolet radiation in the Arctic reduces root biomass and alters microbial community composition but has no effects on microbial biomass

    DEFF Research Database (Denmark)

    Rinnan, R.; Keinänen, M.M.; Kasurinen, A.

    2005-01-01

    We assessed the effects of ambient solar ultraviolet (UV) radiation on below-ground parameters in an arctic heath in north-eastern Greenland. We hypothesized that the current UV fluxes would reduce root biomass and mycorrhizal colonization and that these changes would lead to lower soil microbial...... biomass and altered microbial community composition. These hypotheses were tested on cored soil samples from a UV reduction experiment with three filter treatments (Mylar, 60% UV-B reduction; Lexan, up to 90% UV-B reduction+UV-A reduction; UV transparent Teflon, filter control) and an open control...... treatment in two study sites after 3 years' manipulation. Reduction of both UV-A and UV-B radiation caused over 30% increase in the root biomass of Vaccinium uliginosum, which was the dominant plant species. UV reduction had contrasting effects on ericoid mycorrhizal colonization of V. uliginosum roots...

  13. Applying functional metagenomics to search for novel lignocellulosic enzymes in a microbial consortium derived from a thermophilic composting phase of sugarcane bagasse and cow manure.

    Science.gov (United States)

    Colombo, Lívia Tavares; de Oliveira, Marcelo Nagem Valério; Carneiro, Deisy Guimarães; de Souza, Robson Assis; Alvim, Mariana Caroline Tocantins; Dos Santos, Josenilda Carlos; da Silva, Cynthia Canêdo; Vidigal, Pedro Marcus Pereira; da Silveira, Wendel Batista; Passos, Flávia Maria Lopes

    2016-09-01

    Environments where lignocellulosic biomass is naturally decomposed are sources for discovery of new hydrolytic enzymes that can reduce the high cost of enzymatic cocktails for second-generation ethanol production. Metagenomic analysis was applied to discover genes coding carbohydrate-depleting enzymes from a microbial laboratory subculture using a mix of sugarcane bagasse and cow manure in the thermophilic composting phase. From a fosmid library, 182 clones had the ability to hydrolyse carbohydrate. Sequencing of 30 fosmids resulted in 12 contigs encoding 34 putative carbohydrate-active enzymes belonging to 17 glycosyl hydrolase (GH) families. One third of the putative proteins belong to the GH3 family, which includes β-glucosidase enzymes known to be important in the cellulose-deconstruction process but present with low activity in commercial enzyme preparations. Phylogenetic analysis of the amino acid sequences of seven selected proteins, including three β-glucosidases, showed low relatedness with protein sequences deposited in databases. These findings highlight microbial consortia obtained from a mixture of decomposing biomass residues, such as sugar cane bagasse and cow manure, as a rich resource of novel enzymes potentially useful in biotechnology for saccharification of lignocellulosic substrate.

  14. Microbial keratinases: industrial enzymes with waste management potential.

    Science.gov (United States)

    Verma, Amit; Singh, Hukum; Anwar, Shahbaz; Chattopadhyay, Anirudha; Tiwari, Kapil K; Kaur, Surinder; Dhilon, Gurpreet Singh

    2017-06-01

    Proteases are ubiquitous enzymes that occur in various biological systems ranging from microorganisms to higher organisms. Microbial proteases are largely utilized in various established industrial processes. Despite their numerous industrial applications, they are not efficient in hydrolysis of recalcitrant, protein-rich keratinous wastes which result in environmental pollution and health hazards. This paved the way for the search of keratinolytic microorganisms having the ability to hydrolyze "hard to degrade" keratinous wastes. This new class of proteases is known as "keratinases". Due to their specificity, keratinases have an advantage over normal proteases and have replaced them in many industrial applications, such as nematicidal agents, nitrogenous fertilizer production from keratinous waste, animal feed and biofuel production. Keratinases have also replaced the normal proteases in the leather industry and detergent additive application due to their better performance. They have also been proved efficient in prion protein degradation. Above all, one of the major hurdles of enzyme industrial applications (cost effective production) can be achieved by using keratinous waste biomass, such as chicken feathers and hairs as fermentation substrate. Use of these low cost waste materials serves dual purposes: to reduce the fermentation cost for enzyme production as well as reducing the environmental waste load. The advent of keratinases has given new direction for waste management with industrial applications giving rise to green technology for sustainable development.

  15. Biochemical characterization of thermophilic lignocellulose degrading enzymes and their potential for biomass bioprocessing

    Energy Technology Data Exchange (ETDEWEB)

    Zambare, Vasudeo; Zambare, Archana; Christopher, Lew P. [Center for Bioprocessing Research & Development, South Dakota School of Mines and Technology, Rapid City 57701, SD (United States); Muthukumarappan, Kasiviswanath [Center for Bioprocessing Research & Development, South Dakota State University, Brookings 57007, SD (United States)

    2011-07-01

    . This could have important implications in the enzymatic breakdown of lignocellulosic biomass for the establishment of a robust and cost-efficient process for production of cellulosic ethanol. To the best of our knowledge, this work represents the first report in literature on biochemical characterization of lignocellulose-degrading enzymes from a thermophilic microbial consortium.

  16. Activity assessment of microbial fibrinolytic enzymes.

    Science.gov (United States)

    Kotb, Essam

    2013-08-01

    Conversion of fibrinogen to fibrin inside blood vessels results in thrombosis, leading to myocardial infarction and other cardiovascular diseases. In general, there are four therapy options: surgical operation, intake of antiplatelets, anticoagulants, or fibrinolytic enzymes. Microbial fibrinolytic enzymes have attracted much more attention than typical thrombolytic agents because of the expensive prices and the side effects of the latter. The fibrinolytic enzymes were successively discovered from different microorganisms, the most important among which is the genus Bacillus. Microbial fibrinolytic enzymes, especially those from food-grade microorganisms, have the potential to be developed as functional food additives and drugs to prevent or cure thrombosis and other related diseases. There are several assay methods for these enzymes; this may due to the insolubility of substrate, fibrin. Existing assay methods can be divided into three major groups. The first group consists of assay of fibrinolytic activity with natural proteins as substrates, e.g., fibrin plate methods. The second and third groups of assays are suitable for kinetic studies and are based on the determination of hydrolysis of synthetic peptide esters. This review will deal primarily with the microorganisms that have been reported in literature to produce fibrinolytic enzymes and the first review discussing the methods used to assay the fibrinolytic activity.

  17. Alterations in soil microbial community composition and biomass following agricultural land use change.

    Science.gov (United States)

    Zhang, Qian; Wu, Junjun; Yang, Fan; Lei, Yao; Zhang, Quanfa; Cheng, Xiaoli

    2016-11-04

    The effect of agricultural land use change on soil microbial community composition and biomass remains a widely debated topic. Here, we investigated soil microbial community composition and biomass [e.g., bacteria (B), fungi (F), Arbuscular mycorrhizal fungi (AMF) and Actinomycete (ACT)] using phospholipid fatty acids (PLFAs) analysis, and basal microbial respiration in afforested, cropland and adjacent uncultivated soils in central China. We also investigated soil organic carbon and nitrogen (SOC and SON), labile carbon and nitrogen (LC and LN), recalcitrant carbon and nitrogen (RC and RN), pH, moisture, and temperature. Afforestation averaged higher microbial PLFA biomass compared with cropland and uncultivated soils with higher values in top soils than deep soils. The microbial PLFA biomass was strongly correlated with SON and LC. Higher SOC, SON, LC, LN, moisture and lower pH in afforested soils could be explained approximately 87.3% of total variation of higher total PLFAs. Afforestation also enhanced the F: B ratios compared with cropland. The basal microbial respiration was higher while the basal microbial respiration on a per-unit-PLFA basis was lower in afforested land than adjacent cropland and uncultivated land, suggesting afforestation may increase soil C utilization efficiency and decrease respiration loss in afforested soils.

  18. Microbial Biomass Changes during Decomposition of Plant Residues in a Lixisol

    Directory of Open Access Journals (Sweden)

    Kachaka, SK.

    2003-01-01

    Full Text Available A lixisol was amended with four different alley cropping species: Senna siamea, Leucaena leucocephala, Dactyladenia barteri and Flemingia macrophylla. Soil samples were incubated for 140 days at 25 °C and the soil microbial biomass was determined by the ninhydrin extraction method along the incubation period. The soil microbial biomass values ranged between 80 and 600 mg.kg-1 and followed, in all cases, the decreasing order: Leucaena> Senna> Flemingia> Dactyladenia.

  19. Microbial Community Activity And Plant Biomass Are Insensitive To Passive Warming In A Semiarid Ecosystem

    Science.gov (United States)

    Espinosa, N. J.; Fehmi, J. S.; Rasmussen, C.; Gallery, R. E.

    2017-12-01

    Soil microorganisms drive biogeochemical and nutrient cycling through the production of extracellular enzymes that facilitate organic matter decomposition and the flux of large amounts of carbon dioxide to the atmosphere. Although dryland ecosystems occupy over 40% of land cover and are projected to expand due to climate change, much of our current understanding of these processes comes from mesic temperate ecosystems. Understanding the responses of these globally predominant dryland ecosystems is therefore important yet complicated by co-occurring environmental changes. For example, the widespread and pervasive transition from grass to woody dominated landscapes is changing the hydrology, fire regimes, and carbon storage potential of semiarid ecosystems. In this study, we used a novel passive method of warming to conduct a warming experiment with added plant debris as either woodchip or biochar, to simulate different long-term carbon additions that accompany woody plant encroachment in semiarid ecosystems. The response of heterotrophic respiration, plant biomass, and microbial activity was monitored bi-annually. We hypothesized that the temperature manipulations would have direct and indirect effects on microbial activity. Warmer soils directly reduce the activity of soil extracellular enzymes through denaturation and dehydration of soil pores and indirectly through reducing microbe-available substrates and plant inputs. Overall, reduction in extracellular enzyme activity may reduce decomposition of coarse woody debris and potentially enhance soil carbon storage in semiarid ecosystems. For all seven hydrolytic enzymes examined as well as heterotrophic respiration, there was no consistent or significant response to experimental warming, regardless of seasonal climatic and soil moisture variation. The enzyme results observed here are consistent with the few other experimental results for warming in semiarid ecosystems and indicate that the controls over soil

  20. The contribution of microbial biomass to the adsorption of radioiodide in soils

    International Nuclear Information System (INIS)

    Bors, J.; Martens, R.

    1992-01-01

    The contribution of soil microbial biomass to the sorption and migration of radiodide in soil has been investigated. In two arable soils, a chernozem and a podzol, the numbers of microorganisms were either reduced by biocidal treatment or increased by addition of nutrient sources. Radioiodide ( 125 I - ) adsorption by the pretreated soils was measured, relative to untreated soil samples, in aqueous suspensions containing iodide by estimating the distribution coefficient (K d ) after eight days of incubation. A reduction of biomass to about 10% of its original level drastically decreased adsorption. Elevated levels of microbial biomass (up to 126%) increased adsorption but the increase was not always correlated with biomass level. A closer correlation between soil biomass and adsorption was observed when the concentration of radioiodide in the suspension was increased by several orders of magnitude. Conditions such as anarobiosis and elevated temperatures which are known to influence the activity and survival of microorganisms also exerted an effect on radioiodide sorption. In accordance with the relationship described here between radioiodide adsorption and microbial biomass, migration in water saturated soil columns was influenced by the quantity of microorganisms present. However, high biomass contents obviously caused anaerobic conditions in the system, leading to increased leaching of radioiodide. (author)

  1. The contribution of microbial biomass to the adsorption of radioiodide in soils

    Energy Technology Data Exchange (ETDEWEB)

    Bors, J. (Niedersaechsisches Inst. fuer Radiooekologie, Hannover (Germany)); Martens, R. (Bundesforschungsanstalt fuer Landwirtschaft, Braunschweig (Germany). Inst. fuer Biochemie des Bodens)

    1992-01-01

    The contribution of soil microbial biomass to the sorption and migration of radiodide in soil has been investigated. In two arable soils, a chernozem and a podzol, the numbers of microorganisms were either reduced by biocidal treatment or increased by addition of nutrient sources. Radioiodide ({sup 125}I{sup -}) adsorption by the pretreated soils was measured, relative to untreated soil samples, in aqueous suspensions containing iodide by estimating the distribution coefficient (K{sub d}) after eight days of incubation. A reduction of biomass to about 10% of its original level drastically decreased adsorption. Elevated levels of microbial biomass (up to 126%) increased adsorption but the increase was not always correlated with biomass level. A closer correlation between soil biomass and adsorption was observed when the concentration of radioiodide in the suspension was increased by several orders of magnitude. Conditions such as anarobiosis and elevated temperatures which are known to influence the activity and survival of microorganisms also exerted an effect on radioiodide sorption. In accordance with the relationship described here between radioiodide adsorption and microbial biomass, migration in water saturated soil columns was influenced by the quantity of microorganisms present. However, high biomass contents obviously caused anaerobic conditions in the system, leading to increased leaching of radioiodide. (author).

  2. Modeling adaptation of carbon use efficiency in microbial communities

    Directory of Open Access Journals (Sweden)

    Steven D Allison

    2014-10-01

    Full Text Available In new microbial-biogeochemical models, microbial carbon use efficiency (CUE is often assumed to decline with increasing temperature. Under this assumption, soil carbon losses under warming are small because microbial biomass declines. Yet there is also empirical evidence that CUE may adapt (i.e. become less sensitive to warming, thereby mitigating negative effects on microbial biomass. To analyze potential mechanisms of CUE adaptation, I used two theoretical models to implement a tradeoff between microbial uptake rate and CUE. This rate-yield tradeoff is based on thermodynamic principles and suggests that microbes with greater investment in resource acquisition should have lower CUE. Microbial communities or individuals could adapt to warming by reducing investment in enzymes and uptake machinery. Consistent with this idea, a simple analytical model predicted that adaptation can offset 50% of the warming-induced decline in CUE. To assess the ecosystem implications of the rate-yield tradeoff, I quantified CUE adaptation in a spatially-structured simulation model with 100 microbial taxa and 12 soil carbon substrates. This model predicted much lower CUE adaptation, likely due to additional physiological and ecological constraints on microbes. In particular, specific resource acquisition traits are needed to maintain stoichiometric balance, and taxa with high CUE and low enzyme investment rely on low-yield, high-enzyme neighbors to catalyze substrate degradation. In contrast to published microbial models, simulations with greater CUE adaptation also showed greater carbon storage under warming. This pattern occurred because microbial communities with stronger CUE adaptation produced fewer degradative enzymes, despite increases in biomass. Thus the rate-yield tradeoff prevents CUE adaptation from driving ecosystem carbon loss under climate warming.

  3. Mineralogical impact on long-term patterns of soil nitrogen and phosphorus enzyme activities

    Science.gov (United States)

    Mikutta, Robert; Turner, Stephanie; Meyer-Stüve, Sandra; Guggenberger, Georg; Dohrmann, Reiner; Schippers, Axel

    2014-05-01

    Soil chronosequences provide a unique opportunity to study microbial activity over time in mineralogical diverse soils of different ages. The main objective of this study was to test the effect of mineralogical properties, nutrient and organic matter availability over whole soil pro-files on the abundance and activity of the microbial communities. We focused on microbio-logical processes involved in nitrogen and phosphorus cycling at the 120,000-year Franz Josef soil chronosequence. Microbial abundances (microbial biomass and total cell counts) and enzyme activities (protease, urease, aminopeptidase, and phosphatase) were determined and related to nutrient contents and mineralogical soil properties. Both, microbial abundances and enzyme activities decreased with soil depth at all sites. In the organic layers, microbial biomass and the activities of N-hydrolyzing enzymes showed their maximum at the intermediate-aged sites, corresponding to a high aboveground biomass. In contrast, the phosphatase activity increased with site age. The activities of N-hydrolyzing enzymes were positively correlated with total carbon and nitrogen contents, whereas the phosphatase activity was negatively correlated with the phosphorus content. In the mineral soil, the enzyme activities were generally low, thus reflecting the presence of strongly sorbing minerals. Sub-strate-normalized enzyme activities correlated negatively to clay content as well as poorly crystalline Al and Fe oxyhydroxides, supporting the view that the evolution of reactive sec-ondary mineral phases alters the activity of the microbial communities by constraining sub-strate availability. Our data suggest a strong mineralogical influence on nutrient cycling par-ticularly in subsoil environments.

  4. Technological advances and applications of hydrolytic enzymes for valorization of lignocellulosic biomass.

    Science.gov (United States)

    Manisha; Yadav, Sudesh Kumar

    2017-12-01

    Hydrolytic enzymes are indispensable tools in the production of various foodstuffs, drugs, and consumables owing to their applications in almost every industrial process nowadays. One of the foremost areas of interest involving the use of hydrolytic enzymes is in the transformation of lignocellulosic biomass into value added products. However, limitations of the processes due to inadequate enzyme activity and stability with a narrow range of pH and temperature optima often limit their effective usage. The innovative technologies, involving manipulation of enzyme activity and stability through mutagenesis, genetic engineering and metagenomics lead to a major leap in all the fields using hydrolytic enzymes. This article provides recent advancement towards the isolation and use of microbes for lignocellulosic biomass utilisation, microbes producing the hydrolytic enzymes, the modern age technologies used to manipulate and enhance the hydrolytic enzyme activity and the applications of such enzymes in value added products development from lignocellulosic biomass. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Production of microbial biomass protein by sequential culture fermentation of Arachniotus sp., and Candida utilis

    International Nuclear Information System (INIS)

    Ahmed, S.; Ahmad, F.; Hashmi, A.S.

    2010-01-01

    Sequential culture fermentation by Arachniotus sp. at 35 deg. C for 72 h and followed by Candida utilis fermentation at 35 deg. C for 72 h more resulted in higher production of microbial biomass protein. 6% (w/v) corn stover, 0.0075% CaCl/sub 2/.2H/sub 2/O, 0.005% MgSO/sub 4/.7H/sub 2/O, 0.01% KH/sub 2/PO/sub 4/, C:N ratio of 30:1 and 1% molasses gave higher microbial biomass protein production by the sequential culture fermentation of Arachniotus sp., and C. utilis. The mixed microbial biomass protein produced in the 75-L fermentor contained 16.41%, 23.51%, 10.9%, 12.11% and 0.12% true protein, crude protein, crude fiber, ash and RNA content, respectively. The amino acid profile of final mixed microbial biomass protein showed that it was enriched with essential amino acids. Thus, the potential utilization of corn stover can minimize the cost for growth of these microorganisms and enhance microbial biomass protein production by sequential culture fermentation. (author)

  6. Limitations of ATP as a measure of microbial biomass

    African Journals Online (AJOL)

    limits the use of ATP as a measure of microbial biomass. S. AIr. J. Zool. 1982, 17: 93 - 95. Beraming van die totale .... only micro-organisms present in the experimental culture, but these were later succeeded by a large ... a value of 49: I by the last day of the experiment. Estimates of the total living biomass of the heterotro-.

  7. Effects of elevated nitrogen deposition on soil microbial biomass carbon in major subtropical forests of southern China

    Institute of Scientific and Technical Information of China (English)

    Hui WANG; Jiangming MO; Xiankai LU; Jinghua XUE; Jiong LI; Yunting FANG

    2009-01-01

    The effects of elevated nitrogen deposition on soil microbial biomass carbon (C) and extractable dissolved organic carbon (DOC) in three types of forest of southern China were studied in November, 2004 and June, 2006. Plots were established in a pine forest (PF), a mixed pine and broad-leaved forest (MF) and monsoon evergreen broad-leaved forest (MEBF) in the Dinghushan Nature Reserve. Nitrogen treatments included a control (no N addition), low N (50 kg N/(hm2.a)), medium N (100 kg N/ (hm2. a)) and high N (150 kg N/(hm2. a)). Microbial biomass C and extractable DOC were determined using a chloro-form fumigation-extraction method. Results indicate that microbial biomass C and extractable DOC were higher in June, 2006 than in November, 2004 and higher in the MEBF than in the PF or the MF. The response of soil microbial biomass C and extractable DOC to nitrogen deposition varied depending on the forest type and the level of nitrogen treatment. In the PF or MF forests, no significantly different effects of nitrogen addition were found on soil microbial biomass C and extractable DOC. In the MEBF, however, the soil microbial biomass C generally decreased with increased nitrogen levels and high nitrogen addition significantly reduced soil microbial biomass C. The response of soil extractable DOC to added nitrogen in the MEBF shows the opposite trend to soil microbial biomass C. These results suggest that nitrogen deposition may increase the accumulation of soil organic carbon in the MEBF in the study region.

  8. Development of enzymes and enzyme systems by genetic engineering to convert biomass to sugars

    Science.gov (United States)

    TITLE Development of Enzymes and Enzyme Systems by Genetic Engineering to Convert Biomass to Sugars ABSTRACT Plant cellulosic material is one of the most viable renewable resources for the world’s fuel and chemical feedstock needs. Currently ethanol derived from corn starch is the most common li...

  9. Forest wildfire increases soil microbial biomass C:N:P stoichiometry in long-term effects

    Science.gov (United States)

    Zhou, Xuan

    2017-04-01

    Boreal forest fire strongly influences carbon (C) stock in permafrost soil by thawing permafrost table which accelerated microbe decomposition process. We studied soil microbial biomass stoichiometry in a gradient of four (3 yr, 25 yr, 46 yr and more than 100 yr) ages since fire in Canada boreal forest. Soil microbial biomass (MB) in long-term after fire is significantly higher than in short-term. MB C and nitrogen (N) were mainly dominated by corresponding soil element concentration and inorganic P, while MB phosphorus (P) changes were fully explained by soil N. Fire ages and soil temperature positively increased MB N and P, indicating the negative impact by fire. Microbial C:N:P gradually increased with fire ages from 15:2:1 to 76:6:1 and then drop down to 17:2:1 in the oldest fire ages. The degree of homeostasis of microbial C, N and P are close to 1 indicates non-homoeostasis within microbial elements, while it of C:N:P is close to 8 shows a strong homeostasis within element ratios and proved microbial stoichiometric ratio is not driven by soil element ratios. In conclusion, i) microbial biomass elements highly depends on soil nutrient supply rather than fire ages; ii) wildfire decreased microbial stoichiometry immediate after fire but increased with years after fire (YF) which at least 3 times higher than > 100 fire ages; iii) microbial biomass C, N and P deviated from strict homeostasis but C:N:P ratio reflects stronger homeostasis.

  10. Diversity of Microbial Carbohydrate-Active enZYmes (CAZYmes) Associated with Freshwater and Soil Samples from Caatinga Biome.

    Science.gov (United States)

    Andrade, Ana Camila; Fróes, Adriana; Lopes, Fabyano Álvares Cardoso; Thompson, Fabiano L; Krüger, Ricardo Henrique; Dinsdale, Elizabeth; Bruce, Thiago

    2017-07-01

    Semi-arid and arid areas occupy about 33% of terrestrial ecosystems. However, little information is available about microbial diversity in the semi-arid Caatinga, which represents a unique biome that extends to about 11% of the Brazilian territory and is home to extraordinary diversity and high endemism level of species. In this study, we characterized the diversity of microbial genes associated with biomass conversion (carbohydrate-active enzymes, or so-called CAZYmes) in soil and freshwater of the Caatinga. Our results showed distinct CAZYme profiles in the soil and freshwater samples. Glycoside hydrolases and glycosyltransferases were the most abundant CAZYme families, with glycoside hydrolases more dominant in soil (∼44%) and glycosyltransferases more abundant in freshwater (∼50%). The abundances of individual glycoside hydrolase, glycosyltransferase, and carbohydrate-binding module subfamilies varied widely between soil and water samples. A predominance of glycoside hydrolases was observed in soil, and a higher contribution of enzymes involved in carbohydrate biosynthesis was observed in freshwater. The main taxa associated with the CAZYme sequences were Planctomycetia (relative abundance in soil, 29%) and Alphaproteobacteria (relative abundance in freshwater, 27%). Approximately 5-7% of CAZYme sequences showed low similarity with sequences deposited in non-redundant databases, suggesting putative homologues. Our findings represent a first attempt to describe specific microbial CAZYme profiles for environmental samples. Characterizing these enzyme groups associated with the conversion of carbohydrates in nature will improve our understanding of the significant roles of enzymes in the carbon cycle. We identified a CAZYme signature that can be used to discriminate between soil and freshwater samples, and this signature may be related to the microbial species adapted to the habitat. The data show the potential ecological roles of the CAZYme repertoire and

  11. [Microbial biomass and growth kinetics of microorganisms in chernozem soils under different farm land use modes].

    Science.gov (United States)

    Blagodatskiĭ, S A; Bogomolova, I N; Blagodatskaia, E V

    2008-01-01

    The carbon content of microbial biomass and the kinetic characteristics of microbial respiration response to substrate introduction have been estimated for chernozem soils of different farm lands: arable lands used for 10, 46, and 76 years, mowed fallow land, non-mowed fallow land, and woodland. Microbial biomass and the content of microbial carbon in humus (Cmic/Corg) decreased in the following order: soils under forest cenoses-mowed fallow land-10-year arable land-46- and 75-year arable land. The amount of microbial carbon in the long-plowed horizon was 40% of its content in the upper horizon of non-mowed fallow land. Arable soils were characterized by a lower metabolic diversity of microbial community and by the highest portion of microorganisms able to grow directly on glucose introduced into soil. The effects of different scenarios of carbon sequestration in soil on the reserves and activity of microbial biomass are discussed.

  12. Thermoascus aurantiacus is a promising source of enzymes for biomass deconstruction under thermophilic conditions

    Directory of Open Access Journals (Sweden)

    McClendon Shara D

    2012-07-01

    Full Text Available Abstract Background Thermophilic fungi have attracted increased interest for their ability to secrete enzymes that deconstruct biomass at high temperatures. However, development of thermophilic fungi as enzyme producers for biomass deconstruction has not been thoroughly investigated. Comparing the enzymatic activities of thermophilic fungal strains that grow on targeted biomass feedstocks has the potential to identify promising candidates for strain development. Thielavia terrestris and Thermoascus aurantiacus were chosen for characterization based on literature precedents. Results Thermoascus aurantiacus and Thielavia terrestris were cultivated on various biomass substrates and culture supernatants assayed for glycoside hydrolase activities. Supernatants from both cultures possessed comparable glycoside hydrolase activities when incubated with artificial biomass substrates. In contrast, saccharifications of ionic liquid pretreated switchgrass (Panicum virgatum revealed that T. aurantiacus enzymes released more glucose than T. terrestris enzymes over a range of protein mass loadings and temperatures. Temperature-dependent saccharifications demonstrated that the T. aurantiacus proteins retained higher levels of activity compared to a commercial enzyme mixture sold by Novozymes, Cellic CTec2, at elevated temperatures. Enzymes secreted by T. aurantiacus released glucose at similar protein loadings to CTec2 on dilute acid, ammonia fiber expansion, or ionic liquid pretreated switchgrass. Proteomic analysis of the T. aurantiacus culture supernatant revealed dominant glycoside hydrolases from families 5, 7, 10, and 61, proteins that are key enzymes in commercial cocktails. Conclusions T. aurantiacus produces a complement of secreted proteins capable of higher levels of saccharification of pretreated switchgrass than T. terrestris enzymes. The T. aurantiacus enzymatic cocktail performs at the same level as commercially available enzymatic cocktail for

  13. Thermoascus aurantiacus is a promising source of enzymes for biomass deconstruction under thermophilic conditions.

    Science.gov (United States)

    McClendon, Shara D; Batth, Tanveer; Petzold, Christopher J; Adams, Paul D; Simmons, Blake A; Singer, Steven W

    2012-07-28

    Thermophilic fungi have attracted increased interest for their ability to secrete enzymes that deconstruct biomass at high temperatures. However, development of thermophilic fungi as enzyme producers for biomass deconstruction has not been thoroughly investigated. Comparing the enzymatic activities of thermophilic fungal strains that grow on targeted biomass feedstocks has the potential to identify promising candidates for strain development. Thielavia terrestris and Thermoascus aurantiacus were chosen for characterization based on literature precedents. Thermoascus aurantiacus and Thielavia terrestris were cultivated on various biomass substrates and culture supernatants assayed for glycoside hydrolase activities. Supernatants from both cultures possessed comparable glycoside hydrolase activities when incubated with artificial biomass substrates. In contrast, saccharifications of ionic liquid pretreated switchgrass (Panicum virgatum) revealed that T. aurantiacus enzymes released more glucose than T. terrestris enzymes over a range of protein mass loadings and temperatures. Temperature-dependent saccharifications demonstrated that the T. aurantiacus proteins retained higher levels of activity compared to a commercial enzyme mixture sold by Novozymes, Cellic CTec2, at elevated temperatures. Enzymes secreted by T. aurantiacus released glucose at similar protein loadings to CTec2 on dilute acid, ammonia fiber expansion, or ionic liquid pretreated switchgrass. Proteomic analysis of the T. aurantiacus culture supernatant revealed dominant glycoside hydrolases from families 5, 7, 10, and 61, proteins that are key enzymes in commercial cocktails. T. aurantiacus produces a complement of secreted proteins capable of higher levels of saccharification of pretreated switchgrass than T. terrestris enzymes. The T. aurantiacus enzymatic cocktail performs at the same level as commercially available enzymatic cocktail for biomass deconstruction, without strain development or

  14. Effect of citric acid and microbial phytase on serum enzyme activities ...

    African Journals Online (AJOL)

    Effect of citric acid and microbial phytase on serum enzyme activities and plasma minerals retention in broiler chicks. ... African Journal of Biotechnology ... An experiment was conducted to study the effect of microbial phytase supplementation and citric acid in broiler chicks fed corn-soybean meal base diets on enzyme ...

  15. Annual Removal of Aboveground Plant Biomass Alters Soil Microbial Responses to Warming

    Directory of Open Access Journals (Sweden)

    Kai Xue

    2016-09-01

    Full Text Available Clipping (i.e., harvesting aboveground plant biomass is common in agriculture and for bioenergy production. However, microbial responses to clipping in the context of climate warming are poorly understood. We investigated the interactive effects of grassland warming and clipping on soil properties and plant and microbial communities, in particular, on microbial functional genes. Clipping alone did not change the plant biomass production, but warming and clipping combined increased the C4 peak biomass by 47% and belowground net primary production by 110%. Clipping alone and in combination with warming decreased the soil carbon input from litter by 81% and 75%, respectively. With less carbon input, the abundances of genes involved in degrading relatively recalcitrant carbon increased by 38% to 137% in response to either clipping or the combined treatment, which could weaken long-term soil carbon stability and trigger positive feedback with respect to warming. Clipping alone also increased the abundance of genes for nitrogen fixation, mineralization, and denitrification by 32% to 39%. Such potentially stimulated nitrogen fixation could help compensate for the 20% decline in soil ammonium levels caused by clipping alone and could contribute to unchanged plant biomass levels. Moreover, clipping tended to interact antagonistically with warming, especially with respect to effects on nitrogen cycling genes, demonstrating that single-factor studies cannot predict multifactorial changes. These results revealed that clipping alone or in combination with warming altered soil and plant properties as well as the abundance and structure of soil microbial functional genes. Aboveground biomass removal for biofuel production needs to be reconsidered, as the long-term soil carbon stability may be weakened.

  16. GENPLAT: an automated platform for biomass enzyme discovery and cocktail optimization.

    Science.gov (United States)

    Walton, Jonathan; Banerjee, Goutami; Car, Suzana

    2011-10-24

    The high cost of enzymes for biomass deconstruction is a major impediment to the economic conversion of lignocellulosic feedstocks to liquid transportation fuels such as ethanol. We have developed an integrated high throughput platform, called GENPLAT, for the discovery and development of novel enzymes and enzyme cocktails for the release of sugars from diverse pretreatment/biomass combinations. GENPLAT comprises four elements: individual pure enzymes, statistical design of experiments, robotic pipeting of biomass slurries and enzymes, and automated colorimeteric determination of released Glc and Xyl. Individual enzymes are produced by expression in Pichia pastoris or Trichoderma reesei, or by chromatographic purification from commercial cocktails or from extracts of novel microorganisms. Simplex lattice (fractional factorial) mixture models are designed using commercial Design of Experiment statistical software. Enzyme mixtures of high complexity are constructed using robotic pipeting into a 96-well format. The measurement of released Glc and Xyl is automated using enzyme-linked colorimetric assays. Optimized enzyme mixtures containing as many as 16 components have been tested on a variety of feedstock and pretreatment combinations. GENPLAT is adaptable to mixtures of pure enzymes, mixtures of commercial products (e.g., Accellerase 1000 and Novozyme 188), extracts of novel microbes, or combinations thereof. To make and test mixtures of ˜10 pure enzymes requires less than 100 μg of each protein and fewer than 100 total reactions, when operated at a final total loading of 15 mg protein/g glucan. We use enzymes from several sources. Enzymes can be purified from natural sources such as fungal cultures (e.g., Aspergillus niger, Cochliobolus carbonum, and Galerina marginata), or they can be made by expression of the encoding genes (obtained from the increasing number of microbial genome sequences) in hosts such as E. coli, Pichia pastoris, or a filamentous fungus such

  17. Microbial Activity and Silica Degradation in Rice Straw

    Science.gov (United States)

    Kim, Esther Jin-kyung

    Abundantly available agricultural residues like rice straw have the potential to be feedstocks for bioethanol production. Developing optimized conditions for rice straw deconstruction is a key step toward utilizing the biomass to its full potential. One challenge associated with conversion of rice straw to bioenergy is its high silica content as high silica erodes machinery. Another obstacle is the availability of enzymes that hydrolyze polymers in rice straw under industrially relevant conditions. Microbial communities that colonize compost may be a source of enzymes for bioconversion of lignocellulose to products because composting systems operate under thermophilic and high solids conditions that have been shown to be commercially relevant. Compost microbial communities enriched on rice straw could provide insight into a more targeted source of enzymes for the breakdown of rice straw polysaccharides and silica. Because rice straw is low in nitrogen it is important to understand the impact of nitrogen concentrations on the production of enzyme activity by the microbial community. This study aims to address this issue by developing a method to measure microbial silica-degrading activity and measure the effect of nitrogen amendment to rice straw on microbial activity and extracted enzyme activity during a high-solids, thermophilic incubation. An assay was developed to measure silica-degrading enzyme or silicase activity. This process included identifying methods of enzyme extraction from rice straw, identifying a model substrate for the assay, and optimizing measurement techniques. Rice straw incubations were conducted with five different levels of nitrogen added to the biomass. Microbial activity was measured by respiration and enzyme activity. A microbial community analysis was performed to understand the shift in community structure with different treatments. With increased levels of nitrogen, respiration and cellulose and hemicellulose degrading activity

  18. Microbial biomass in faeces of dairy cows affected by a nitrogen deficient diet.

    Science.gov (United States)

    Jost, Daphne Isabel; Aschemann, Martina; Lebzien, Peter; Joergensen, Rainer Georg; Sundrum, Albert

    2013-04-01

    Since more than half of the faecal nitrogen (N) originates from microbial N, the objective of the study was to develop a method for quantitatively detecting microbial biomass and portion of living microorganisms in dairy cattle faeces, including bacteria, fungi and archaea. Three techniques were tested: (1) the chloroform fumigation extraction (CFE) method, (2) detection of the fungal cell-membrane component ergosterol and (3) analysis of the cell wall components fungal glucosamine and bacterial muramic acid. In a second step, an N deficient (ND) and an N balanced (NB) diets were compared with respect to the impacts on faecal C and N fractions, microbial indices and digestibility. The mean values of microbial biomass C and N concentrations averaged around 37 and 4.9 mg g(-1) DM, respectively. Ergosterol, together with fungal glucosamine and bacterial muramic acid, revealed a 25% fungal C in relation to the total microbial C content in dairy cattle faeces. Changes in ruminal N supply showed significant effects on faecal composition. Faecal concentrations of NDF, hemicelluloses and undigested dietary N and the total C/N ratio were significantly higher in ND treatment compared to the NB treatment. N deficiency was reflected also by a higher microbial biomass C/N ratio. It was concluded that the assessment of microbial indices provides valuable information with respect to diet effects on faecal composition and the successive decomposition. Further studies should be conducted to explore the potentials for minimising nutrient losses from faeces.

  19. The survival strategy of the soil microbial biomass

    Science.gov (United States)

    Brookes, Philip; Kemmitt, Sarah; Dungait, Jennifer; Xu, Jianming

    2014-05-01

    The soil microbial biomass (biomass) is defined as the sum of the masses of all soil microorganisms > 5000 µm3 (e.g. fungi, bacteria, protozoa, yeasts, actinomycetes and algae). Typically comprising about 1 to 3 % of total soil organic matter (SOM), the biomass might be though to live in a highly substrate-rich environment. However, the SOM is, normally, only exceedingly slowly available to the biomass. However the biomass can survive for months or even years on this meagre energy source. Not surprisingly, therefore, the biomass exhibits many features typical of a dormant or resting population. These include a very low rate of basal and specific respiration, a slow rate of cell division (about once every six months on average) and slow turnover rate. These are clearly adaptations to existing in an environment where substrate availability is very low. Yet, paradoxically, the biomass, in soils worldwide, has an adenosine triphosphate (ATP) concentration (around 10 to 12 µmol ATP g-1 biomass C), and an Adenylate Energy Charge (AEC = [(ATP) + (0.5 ADP)]/[(ATP)+(ADP) + (AMP)]) which are typical of microorganisms growing exponentially in a chemostat. This sets us several questions. Firstly, under the condition of extremely limited substrate availability in soil, why does the biomass not mainly exist as spores, becoming active, by increasing both its ATP concentration and AEC, when substrate (plant and animal residues) becomes available? We surmise that a spore strategy may put organisms at a competitive disadvantage, compared to others which are prepared to invest energy, maintaining high ATP and ATP, to take advantage of a 'food event' as soon as it becomes available. Secondly, since SOM is available (although only very slowly) to the biomass, why have some groups not evolved the ability to mineralize it faster, obtain more energy, and so gain a competitive advantage? We believe that the reason why organisms do not use this strategy is, simply, that they cannot. Our

  20. Research and Application of Marine Microbial Enzymes: Status and Prospects

    Science.gov (United States)

    Zhang, Chen; Kim, Se-Kwon

    2010-01-01

    Over billions of years, the ocean has been regarded as the origin of life on Earth. The ocean includes the largest range of habitats, hosting the most life-forms. Competition amongst microorganisms for space and nutrients in the marine environment is a powerful selective force, which has led to evolution. The evolution prompted the marine microorganisms to generate multifarious enzyme systems to adapt to the complicated marine environments. Therefore, marine microbial enzymes can offer novel biocatalysts with extraordinary properties. This review deals with the research and development work investigating the occurrence and bioprocessing of marine microbial enzymes. PMID:20631875

  1. Microbial genetic engineering and enzyme technology

    Energy Technology Data Exchange (ETDEWEB)

    Hollenberg, C.P.; Sahm, H.

    1987-01-01

    In a series of up-to-date contributions BIOTEC 1 has experts discussing the current topics in microbial gene technology and enzyme technology and speculating on future developments. Bacterial and yeast systems for the production of interferons, growth hormone or viral antigenes are described as well as the impact of gene technology on plants. Exciting is the prospect of degrading toxic compounds in our environment by microorganisms tuned in the laboratory. Enzymes are the most effective catalysts we know. They exhibit a very high substrate- and stereospecificity. These properties make enzymes extremely attractive as industrial catalysts, leading to new production processes that are non-polluting and save both energy and raw materials. (orig.) With 135 figs., 36 tabs.

  2. Trichoderma Biofertilizer Links to Altered Soil Chemistry, Altered Microbial Communities, and Improved Grassland Biomass

    Directory of Open Access Journals (Sweden)

    Fengge Zhang

    2018-04-01

    Full Text Available In grasslands, forage and livestock production results in soil nutrient deficits as grasslands typically receive no nutrient inputs, leading to a loss of grassland biomass. The application of mature compost has been shown to effectively increase grassland nutrient availability. However, research on fertilization regime influence and potential microbial ecological regulation mechanisms are rarely conducted in grassland soil. We conducted a two-year experiment in meadow steppe grasslands, focusing on above- and belowground consequences of organic or Trichoderma biofertilizer applications and potential soil microbial ecological mechanisms underlying soil chemistry and microbial community responses. Grassland biomass significantly (p = 0.019 increased following amendment with 9,000 kg ha−1 of Trichoderma biofertilizer (composted cattle manure + inoculum compared with other assessed organic or biofertilizer rates, except for BOF3000 (fertilized with 3,000 kg ha−1 biofertilizer. This rate of Trichoderma biofertilizer treatment increased soil antifungal compounds that may suppress pathogenic fungi, potentially partially responsible for improved grassland biomass. Nonmetric multidimensional scaling (NMDS revealed soil chemistry and fungal communities were all separated by different fertilization regime. Trichoderma biofertilizer (9,000 kg ha−1 increased relative abundances of Archaeorhizomyces and Trichoderma while decreasing Ophiosphaerella. Trichoderma can improve grassland biomass, while Ophiosphaerella has the opposite effect as it may secrete metabolites causing grass necrosis. Correlations between soil properties and microbial genera showed plant-available phosphorus may influence grassland biomass by increasing Archaeorhizomyces and Trichoderma while reducing Ophiosphaerella. According to our structural equation modeling (SEM, Trichoderma abundance was the primary contributor to aboveground grassland biomass. Our results suggest Trichoderma

  3. Trichoderma Biofertilizer Links to Altered Soil Chemistry, Altered Microbial Communities, and Improved Grassland Biomass.

    Science.gov (United States)

    Zhang, Fengge; Huo, Yunqian; Cobb, Adam B; Luo, Gongwen; Zhou, Jiqiong; Yang, Gaowen; Wilson, Gail W T; Zhang, Yingjun

    2018-01-01

    In grasslands, forage and livestock production results in soil nutrient deficits as grasslands typically receive no nutrient inputs, leading to a loss of grassland biomass. The application of mature compost has been shown to effectively increase grassland nutrient availability. However, research on fertilization regime influence and potential microbial ecological regulation mechanisms are rarely conducted in grassland soil. We conducted a two-year experiment in meadow steppe grasslands, focusing on above- and belowground consequences of organic or Trichoderma biofertilizer applications and potential soil microbial ecological mechanisms underlying soil chemistry and microbial community responses. Grassland biomass significantly ( p = 0.019) increased following amendment with 9,000 kg ha -1 of Trichoderma biofertilizer (composted cattle manure + inoculum) compared with other assessed organic or biofertilizer rates, except for BOF3000 (fertilized with 3,000 kg ha -1 biofertilizer). This rate of Trichoderma biofertilizer treatment increased soil antifungal compounds that may suppress pathogenic fungi, potentially partially responsible for improved grassland biomass. Nonmetric multidimensional scaling (NMDS) revealed soil chemistry and fungal communities were all separated by different fertilization regime. Trichoderma biofertilizer (9,000 kg ha -1 ) increased relative abundances of Archaeorhizomyces and Trichoderma while decreasing Ophiosphaerella . Trichoderma can improve grassland biomass, while Ophiosphaerella has the opposite effect as it may secrete metabolites causing grass necrosis. Correlations between soil properties and microbial genera showed plant-available phosphorus may influence grassland biomass by increasing Archaeorhizomyces and Trichoderma while reducing Ophiosphaerella . According to our structural equation modeling (SEM), Trichoderma abundance was the primary contributor to aboveground grassland biomass. Our results suggest Trichoderma

  4. Microbial surface displayed enzymes based biofuel cell utilizing degradation products of lignocellulosic biomass for direct electrical energy.

    Science.gov (United States)

    Fan, Shuqin; Hou, Chuantao; Liang, Bo; Feng, Ruirui; Liu, Aihua

    2015-09-01

    In this work, a bacterial surface displaying enzyme based two-compartment biofuel cell for the direct electrical energy conversion from degradation products of lignocellulosic biomass is reported. Considering that the main degradation products of the lignocellulose are glucose and xylose, xylose dehydrogenase (XDH) displayed bacteria (XDH-bacteria) and glucose dehydrogenase (GDH) displayed bacteria (GDH-bacteria) were used as anode catalysts in anode chamber with methylene blue as electron transfer mediator. While the cathode chamber was constructed with laccase/multi-walled-carbon nanotube/glassy-carbon-electrode. XDH-bacteria exhibited 1.75 times higher catalytic efficiency than GDH-bacteria. This assembled enzymatic fuel cell exhibited a high open-circuit potential of 0.80 V, acceptable stability and energy conversion efficiency. Moreover, the maximum power density of the cell could reach 53 μW cm(-2) when fueled with degradation products of corn stalk. Thus, this finding holds great potential to directly convert degradation products of biomass into electrical energy. Copyright © 2015 Elsevier Ltd. All rights reserved.

  5. Shotgun Approach to Increasing Enzymatic Saccharification Yields of Ammonia Fiber Expansion Pretreated Cellulosic Biomass

    International Nuclear Information System (INIS)

    Chundawat, Shishir P. S.; Uppugundla, Nirmal; Gao, Dahai; Curran, Paul G.; Balan, Venkatesh; Dale, Bruce E.

    2017-01-01

    Most cellulolytic enzyme blends, either procured from a commercial vendor or isolated from a single cellulolytic microbial secretome, do not efficiently hydrolyze ammonia-pretreated (e.g., ammonia fiber expansion, AFEX) lignocellulosic agricultural crop residues like corn stover to fermentable sugars. Typically reported commercial enzyme loading (30–100 mg protein/g glucan) necessary to achieve >90% total hydrolysis yield (to monosaccharides) for AFEX-treated biomass, within a short saccharification time frame (24–48 h), is economically unviable. Unlike acid-based pretreatments, AFEX retains most of the hemicelluloses in the biomass and therefore requires a more complex suite of enzymes for efficient hydrolysis of cellulose and hemicellulose at industrially relevant high solids loadings. One strategy to reduce enzyme dosage while improving cocktail effectiveness for AFEX-treated biomass has been to use individually purified enzymes to determine optimal enzyme combinations to maximize hydrolysis yields. However, this approach is limited by the selection of heterologous enzymes available or the labor required for isolating low-abundance enzymes directly from the microbial secretomes. Here, we show that directly blending crude cellulolytic and hemicellulolytic enzymes-rich microbial secretomes can maximize specific activity on AFEX-treated biomass without having to isolate individual enzymes. Fourteen commercially available cellulolytic and hemicellulolytic enzymes were procured from leading enzyme companies (Novozymes ® , Genencor ® , and Biocatalysts ® ) and were mixed together to generate several hundred unique cocktail combinations. The mixtures were assayed for activity on AFEX-treated corn stover (AFEX-CS) using a previously established high-throughput methodology. The optimal enzyme blend combinations identified from these screening assays were enriched in various low-abundance hemicellulases and accessory enzymes typically absent in most commercial

  6. Shotgun Approach to Increasing Enzymatic Saccharification Yields of Ammonia Fiber Expansion Pretreated Cellulosic Biomass

    Energy Technology Data Exchange (ETDEWEB)

    Chundawat, Shishir P. S., E-mail: shishir.chundawat@rutgers.edu [Department of Chemical and Biochemical Engineering, Rutgers-State University of New Jersey, Piscataway, NJ (United States); Uppugundla, Nirmal; Gao, Dahai [Department of Chemical Engineering and Materials Science, Michigan State University, East Lansing, MI (United States); Curran, Paul G. [Center for Statistical Training and Consulting (CSTAT), Michigan State University, East Lansing, MI (United States); Balan, Venkatesh; Dale, Bruce E. [Department of Chemical Engineering and Materials Science, Michigan State University, East Lansing, MI (United States)

    2017-05-10

    Most cellulolytic enzyme blends, either procured from a commercial vendor or isolated from a single cellulolytic microbial secretome, do not efficiently hydrolyze ammonia-pretreated (e.g., ammonia fiber expansion, AFEX) lignocellulosic agricultural crop residues like corn stover to fermentable sugars. Typically reported commercial enzyme loading (30–100 mg protein/g glucan) necessary to achieve >90% total hydrolysis yield (to monosaccharides) for AFEX-treated biomass, within a short saccharification time frame (24–48 h), is economically unviable. Unlike acid-based pretreatments, AFEX retains most of the hemicelluloses in the biomass and therefore requires a more complex suite of enzymes for efficient hydrolysis of cellulose and hemicellulose at industrially relevant high solids loadings. One strategy to reduce enzyme dosage while improving cocktail effectiveness for AFEX-treated biomass has been to use individually purified enzymes to determine optimal enzyme combinations to maximize hydrolysis yields. However, this approach is limited by the selection of heterologous enzymes available or the labor required for isolating low-abundance enzymes directly from the microbial secretomes. Here, we show that directly blending crude cellulolytic and hemicellulolytic enzymes-rich microbial secretomes can maximize specific activity on AFEX-treated biomass without having to isolate individual enzymes. Fourteen commercially available cellulolytic and hemicellulolytic enzymes were procured from leading enzyme companies (Novozymes{sup ®}, Genencor{sup ®}, and Biocatalysts{sup ®}) and were mixed together to generate several hundred unique cocktail combinations. The mixtures were assayed for activity on AFEX-treated corn stover (AFEX-CS) using a previously established high-throughput methodology. The optimal enzyme blend combinations identified from these screening assays were enriched in various low-abundance hemicellulases and accessory enzymes typically absent in most

  7. Metal impacts on microbial biomass in the anoxic sediments of a contaminated lake

    Energy Technology Data Exchange (ETDEWEB)

    Gough, Heidi L.; Dahl, Amy L.; Nolan, Melissa A.; Gaillard, Jean-Francois; Stahl, David A.

    2008-04-26

    Little is known about the long-term impacts of metal contamination on the microbiota of anoxic lake sediments. In this study, we examined microbial biomass and metals (arsenic, cadmium, chromium, copper, iron, lead, manganese, and zinc) in the sediments of Lake DePue, a backwater lake located near a former zinc smelter. Sediment core samples were examined using two independent measures for microbial biomass (total microscopic counts and total phospholipid-phosphate concentrations), and for various fractions of each metal (pore water extracts, sequential extractions, and total extracts of all studied metals and zinc speciation by X-ray absorption fine structure (XAFS). Zinc concentrations were up to 1000 times higher than reported for sediments in the adjacent Illinois River, and ranged from 21,400 mg/kg near the source to 1,680 mg/kg near the river. However, solid metal fractions were not well correlated with pore water concentrations, and were not good predictors of biomass concentrations. Instead, biomass, which varied among sites by as much as two-times, was inversely correlated with concentrations of pore water zinc and arsenic as established by multiple linear regression. Monitoring of other parameters known to naturally influence biomass in sediments (e.g., organic carbon concentrations, nitrogen concentrations, pH, sediment texture, and macrophytes) revealed no differences that could explain observed biomass trends. This study provides strong support for control of microbial abundance by pore water metal concentrations in contaminated freshwater sediments.

  8. Membrane bioreactor biomass characteristics and microbial yield at ...

    African Journals Online (AJOL)

    In this study, a laboratory-scale MBR and SBR were operated in parallel and at very low MCRTs (3 d, 2 d, 1 d and 0.5 d) to assess the relative bioreactor performance, biomass characteristics, and microbial yield. This study confirmed that the MBR maintains higher solids levels and better overall effluent quality than ...

  9. Effects of inorganic and organic amendment on soil chemical properties, enzyme activities, microbial community and soil quality in yellow clayey soil.

    Directory of Open Access Journals (Sweden)

    Zhanjun Liu

    Full Text Available Understanding the effects of external organic and inorganic components on soil fertility and quality is essential for improving low-yielding soils. We conducted a field study over two consecutive rice growing seasons to investigate the effect of applying chemical fertilizer (NPK, NPK plus green manure (NPKG, NPK plus pig manure (NPKM, and NPK plus straw (NPKS on the soil nutrient status, enzyme activities involved in C, N, P, and S cycling, microbial community and rice yields of yellow clayey soil. Results showed that the fertilized treatments significantly improved rice yields over the first three experimental seasons. Compared with the NPK treatment, organic amendments produced more favorable effects on soil productivity. Notably, the NPKM treatment exhibited the highest levels of nutrient availability, microbial biomass carbon (MBC, activities of most enzymes and the microbial community. This resulted in the highest soil quality index (SQI and rice yield, indicating better soil fertility and quality. Significant differences in enzyme activities and the microbial community were observed among the treatments, and redundancy analysis showed that MBC and available N were the key determinants affecting the soil enzyme activities and microbial community. The SQI score of the non-fertilized control (0.72 was comparable to that of the NPK (0.77, NPKG (0.81 and NPKS (0.79 treatments but significantly lower compared with NPKM (0.85. The significant correlation between rice yield and SQI suggests that SQI can be a useful to quantify soil quality changes caused by different agricultural management practices. The results indicate that application of NPK plus pig manure is the preferred option to enhance SOC accumulation, improve soil fertility and quality, and increase rice yield in yellow clayey soil.

  10. Radiocesium storage in soil microbial biomass of undisturbed alpine meadow soils and its relation to 137Cs soil-plant transfer

    International Nuclear Information System (INIS)

    Stemmer, Michael; Hromatka, Angelika; Lettner, Herbert; Strebl, Friederike

    2005-01-01

    This study focuses on radiocesium storage in soil microbial biomass of undisturbed alpine meadow sites and its relation to the soil-to-plant transfer. Soil and plant samples were taken in August 1999 from an altitude transect (800-1600 m.a.s.l.) at Gastein valley, Austria. Soil samples were subdivided into 3-cm layers for analyses of total, K 2 SO 4 -extractable and microbially stored 137 Cs. Microbial biomass was measured by the fumigation extraction method, and fungal biomass was quantified using ergosterol as biomarker molecule. In general, the quantity of 137 Cs stored in the living soil microbial biomass was relatively small. At the high-altitude meadows, showing high amounts of fungal biomass, microbially stored 137 Cs amounted to 0.64 ± 0.14 kBq m -2 which corresponds to about 1.2-2.7% of the total 137 Cs soil inventory. At lower altitudes, microbial 137 Cs content was distinctly smaller and in most cases not measurable at all using the fumigation extraction method. However, a positive correlation between the observed soil-to-plant aggregated transfer factor, microbially stored 137 Cs and fungal biomass was found, which indicates a possible role of fungal biomass in the storage and turnover of 137 Cs in soils and in the 137 Cs uptake by plants

  11. Enzyme and methodology for the treatment of a biomass

    Science.gov (United States)

    Thompson, Vicki S.; Thompson, David N.; Schaller, Kastli D.; Apel, William A.

    2010-06-01

    An enzyme isolated from an extremophilic microbe, and a method for utilizing same is described, and wherein the enzyme displays optimum enzymatic activity at a temperature of greater than about 80.degree. C., and a pH of less than about 2, and further may be useful in methodology including pretreatment of a biomass so as to facilitate the production of an end product.

  12. High levels of maize in broiler diets with or without microbial enzyme ...

    African Journals Online (AJOL)

    Over the feeding period (21 d), there was an increase in feed intake as maize inclusion level (MIL) increased in diets, while supplementation with microbial enzyme improved feed intake only in the MM diet. There was an improvement in live weight (LW) in chickens with increased MIL in their diets. The microbial enzyme ...

  13. Ecological Insights into the Dynamics of Plant Biomass-Degrading Microbial Consortia.

    Science.gov (United States)

    Jiménez, Diego Javier; Dini-Andreote, Francisco; DeAngelis, Kristen M; Singer, Steven W; Salles, Joana Falcão; van Elsas, Jan Dirk

    2017-10-01

    Plant biomass (PB) is an important resource for biofuel production. However, the frequent lack of efficiency of PB saccharification is still an industrial bottleneck. The use of enzyme cocktails produced from PB-degrading microbial consortia (PB-dmc) is a promising approach to optimize this process. Nevertheless, the proper use and manipulation of PB-dmc depends on a sound understanding of the ecological processes and mechanisms that exist in these communities. This Opinion article provides an overview of arguments as to how spatiotemporal nutritional fluxes influence the successional dynamics and ecological interactions (synergism versus competition) between populations in PB-dmc. The themes of niche occupancy, 'sugar cheaters', minimal effective consortium, and the Black Queen Hypothesis are raised as key subjects that foster our appraisal of such systems. Here we provide a conceptual framework that describes the critical topics underpinning the ecological basis of PB-dmc, giving a solid foundation upon which further prospective experimentation can be developed. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Effects of vegetation type on microbial biomass carbon and nitrogen in subalpine mountain forest soils.

    Science.gov (United States)

    Ravindran, Anita; Yang, Shang-Shyng

    2015-08-01

    Microbial biomass plays an important role in nutrient transformation and conservation of forest and grassland ecosystems. The objective of this study was to determine the microbial biomass among three vegetation types in subalpine mountain forest soils of Taiwan. Tatachia is a typical high-altitude subalpine temperate forest ecosystem in Taiwan with an elevation of 1800-3952 m and consists of three vegetation types: spruce, hemlock, and grassland. Three plots were selected in each vegetation type. Soil samples were collected from the organic layer, topsoil, and subsoil. Microbial biomass carbon (Cmic) was determined by the chloroform fumigation-extraction method, and microbial biomass nitrogen (Nmic) was determined from the total nitrogen (Ntot) released during fumigation-extraction. Bacteria, actinomycetes, fungi, cellulolytic microbes, phosphate-solubilizing microbes, and nitrogen-fixing microbes were also counted. The Cmic and Nmic were highest in the surface soil and declined with the soil depth. These were also highest in spruce soils, followed by in hemlock soils, and were lowest in grassland soils. Cmic and Nmic had the highest values in the spring season and the lowest values in the winter season. Cmic and Nmic had significantly positive correlations with total organic carbon (Corg) and Ntot. Contributions of Cmic and Nmic, respectively, to Corg and Ntot indicated that the microbial biomass was immobilized more in spruce and hemlock soils than in grassland soils. Microbial populations of the tested vegetation types decreased with increasing soil depth. Cmic and Nmic were high in the organic layer and decreased with the depth of layers. These values were higher for spruce and hemlock soils than for grassland soils. Positive correlations were observed between Cmic and Nmic and between Corg and Ntot. Copyright © 2014. Published by Elsevier B.V.

  15. Afforestation impacts microbial biomass and its natural {sup 13}C and {sup 15}N abundance in soil aggregates in central China

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Junjun; Zhang, Qian; Yang, Fan; Lei, Yao; Zhang, Quanfa; Cheng, Xiaoli, E-mail: xlcheng@fudan.edu.cn

    2016-10-15

    We investigated soil microbial biomass and its natural abundance of δ{sup 13}C and δ{sup 15}N in aggregates (> 2000 μm, 250–2000 μm, 53–250 μm and < 53 μm) of afforested (implementing woodland and shrubland plantations) soils, adjacent croplands and open area (i.e., control) in the Danjiangkou Reservoir area of central China. The afforested soils averaged higher microbial biomass carbon (MBC) and nitrogen (MBN) levels in all aggregates than in open area and cropland, with higher microbial biomass in micro-aggregates (< 250 μm) than in macro-aggregates (> 2000 μm). The δ{sup 13}C of soil microbial biomass was more enriched in woodland soils than in other land use types, while δ{sup 15}N of soil microbial biomass was more enriched compared with that of organic soil in all land use types. The δ{sup 13}C and δ{sup 15}N of microbial biomass were positively correlated with the δ{sup 13}C and δ{sup 15}N of organic soil across aggregates and land use types, whereas the {sup 13}C and {sup 15}N enrichment of microbial biomass exhibited linear decreases with the corresponding C:N ratio of organic soil. Our results suggest that shifts in the natural {sup 13}C and {sup 15}N abundance of microbial biomass reflect changes in the stabilization and turnover of soil organic matter (SOM) and thereby imply that afforestation can greatly impact SOM accumulation over the long-term. - Highlights: • Afforested soils averaged higher microbial biomass in all aggregates than cropland. • Microbial biomass was higher in micro-aggregates than in macro-aggregates. • δ{sup 13}C and δ{sup 15}N of microbe positively correlated with δ{sup 13}C and δ{sup 15}N of organic soil. • {sup 13}C and {sup 15}N enrichment of microbe was negatively related to with soil C:N ratio.

  16. Effect of monospecific and mixed sea-buckthorn (Hippophae rhamnoides plantations on the structure and activity of soil microbial communities.

    Directory of Open Access Journals (Sweden)

    Xuan Yu

    Full Text Available This study aims to evaluate the effect of different afforestation models on soil microbial composition in the Loess Plateau in China. In particular, we determined soil physicochemical properties, enzyme activities, and microbial community structures in the top 0 cm to 10 cm soil underneath a pure Hippophae rhamnoides (SS stand and three mixed stands, namely, H. rhamnoides and Robinia pseucdoacacia (SC, H. rhamnoides and Pinus tabulaeformis (SY, and H. rhamnoides and Platycladus orientalis (SB. Results showed that total organic carbon (TOC, total nitrogen, and ammonium (NH4(+ contents were higher in SY and SB than in SS. The total microbial biomass, bacterial biomass, and Gram+ biomass of the three mixed stands were significantly higher than those of the pure stand. However, no significant difference was found in fungal biomass. Correlation analysis suggested that soil microbial communities are significantly and positively correlated with some chemical parameters of soil, such as TOC, total phosphorus, total potassium, available phosphorus, NH4(+ content, nitrate content (NH3(-, and the enzyme activities of urease, peroxidase, and phosphatase. Principal component analysis showed that the microbial community structures of SB and SS could clearly be discriminated from each other and from the others, whereas SY and SC were similar. In conclusion, tree species indirectly but significantly affect soil microbial communities and enzyme activities through soil physicochemical properties. In addition, mixing P. tabulaeformis or P. orientalis in H. rhamnoides plantations is a suitable afforestation model in the Loess Plateau, because of significant positive effects on soil nutrient conditions, microbial community, and enzyme activities over pure plantations.

  17. Soil plus root respiration and microbial biomass following water, nitrogen, and phosphorus application at a high arctic semi desert

    DEFF Research Database (Denmark)

    Illeris, Lotte; Michelsen, Anders; Jonasson, Sven Evert

    2003-01-01

    CO2 emmision, Decomposition, Microbial biomass carbon, Soil organic matter, Tundra, Water and nutrient limitation......CO2 emmision, Decomposition, Microbial biomass carbon, Soil organic matter, Tundra, Water and nutrient limitation...

  18. Recent trends in ionic liquid (IL) tolerant enzymes and microorganisms for biomass conversion.

    Science.gov (United States)

    Portillo, Maria Del Carmen; Saadeddin, Anas

    2015-01-01

    Second generation biofuel production depends on lignocellulosic (LC) biomass transformation into simple sugars and their subsequent fermentation into alcohols. However, the main obstacle in this process is the efficient breakdown of the recalcitrant cellulose to sugar monomers. Hence, efficient feedstock pretreatment and hydrolysis are necessary to produce a cost effective biofuel. Recently, ionic liquids (ILs) have been recognized as a promising solvent able to dissolve different biomass feedstocks, providing higher sugar yields. However, most of the hydrolytic enzymes and microorganisms are inactivated, completely or partially, in the presence of even low concentrations of IL, making necessary the discovery of novel hydrolytic enzymes and fermentative microorganisms that are tolerant to ILs. In this review, the current state and the challenges of using ILs as a pretreatment of LC biomass was evaluated, underlining the advances in the discovery and identification of new IL-tolerant enzymes and microorganisms that could improve the bioprocessing of biomass to fuels and chemicals.

  19. Microbial plankton communities in the coastal southeastern Black Sea: biomass, composition and trophic interactions

    Directory of Open Access Journals (Sweden)

    Ulgen Aytan

    2018-04-01

    Full Text Available Summary: We investigated biomass and composition of the pico-, nano- and microplankton communities in a coastal station of the southeastern Black Sea during 2011. We also examined trophic interactions within these communities from size-fractionated dilution experiments in February, June and December. Autotrophic and heterotrophic biomasses showed similar seasonal trends, with a peak in June, but heterotrophs dominated throughout the year. Autotrophic biomass was mainly comprised by nanoflagellates and diatoms in the first half of the year, and by dinoflagellates and Synechococcus spp. in the second half. Heterotrophic biomass was mostly dominated by heterotrophic bacteria, followed by nanoflagellates and microzooplankton. Dilution experiments suggest that nano- and microzooplankton were significant consumers of autotrophs and heterotrophic bacteria. More than 100% of bacterial production was consumed by grazers in all experiments, while 46%, 21% and 30% of daily primary production were consumed in February, June and December, respectively. In February, autotrophs were the main carbon source, but in December, it was heterotrophic bacteria. An intermediate situation was observed in June, with similar carbon flows from autotrophs and heterotrophic bacteria. Size-fraction dilution experiments suggested that heterotrophic nanoflagellates are an important link between the high heterotrophic bacterial biomass and microzooplankton. In summary, these results indicate that nano- and microzooplankton were responsible for comprising a significant fraction of total microbial plankton biomass, standing stocks, growth and grazing processes. This suggests that in 2011, the microbial food web was an important compartment of the planktonic food web in the coastal southeastern Black Sea. Keywords: Phytoplankton, Microzooplankton, Carbon biomass, Microbial food web, Grazing, Black Sea

  20. Microbial chemical factories: recent advances in pathway engineering for synthesis of value added chemicals.

    Science.gov (United States)

    Dhamankar, Himanshu; Prather, Kristala L J

    2011-08-01

    The dwindling nature of petroleum and other fossil reserves has provided impetus towards microbial synthesis of fuels and value added chemicals from biomass-derived sugars as a renewable resource. Microbes have naturally evolved enzymes and pathways that can convert biomass into hundreds of unique chemical structures, a property that can be effectively exploited for their engineering into Microbial Chemical Factories (MCFs). De novo pathway engineering facilitates expansion of the repertoire of microbially synthesized compounds beyond natural products. In this review, we visit some recent successes in such novel pathway engineering and optimization, with particular emphasis on the selection and engineering of pathway enzymes and balancing of their accessory cofactors. Copyright © 2011 Elsevier Ltd. All rights reserved.

  1. Looking inside the box: using Raman microspectroscopy to deconstruct microbial biomass stoichiometry one cell at a time

    Science.gov (United States)

    Hall, Edward K.; Singer, Gabriel A.; Pölzl, Marvin; Hämmerle, Ieda; Schwarz, Christian; Daims, Holger; Maixner, Frank; Battin, Tom J.

    2011-01-01

    Stoichiometry of microbial biomass is a key determinant of nutrient recycling in a wide variety of ecosystems. However, little is known about the underlying causes of variance in microbial biomass stoichiometry. This is primarily because of technological constraints limiting the analysis of macromolecular composition to large quantities of microbial biomass. Here, we use Raman microspectroscopy (MS), to analyze the macromolecular composition of single cells of two species of bacteria grown on minimal media over a wide range of resource stoichiometry. We show that macromolecular composition, determined from a subset of identified peaks within the Raman spectra, was consistent with macromolecular composition determined using traditional analytical methods. In addition, macromolecular composition determined by Raman MS correlated with total biomass stoichiometry, indicating that analysis with Raman MS included a large proportion of a cell's total macromolecular composition. Growth phase (logarithmic or stationary), resource stoichiometry and species identity each influenced each organism's macromolecular composition and thus biomass stoichiometry. Interestingly, the least variable peaks in the Raman spectra were those responsible for differentiation between species, suggesting a phylogenetically specific cellular architecture. As Raman MS has been previously shown to be applicable to cells sampled directly from complex environments, our results suggest Raman MS is an extremely useful application for evaluating the biomass stoichiometry of environmental microorganisms. This includes the ability to partition microbial biomass into its constituent macromolecules and increase our understanding of how microorganisms in the environment respond to resource heterogeneity.

  2. Microbial Functional Diversity, Biomass and Activity as Affected by Soil Surface Mulching in a Semiarid Farmland.

    Directory of Open Access Journals (Sweden)

    Yufang Shen

    Full Text Available Mulching is widely used to increase crop yield in semiarid regions in northwestern China, but little is known about the effect of different mulching systems on the microbial properties of the soil, which play an important role in agroecosystemic functioning and nutrient cycling. Based on a 4-year spring maize (Zea mays L. field experiment at Changwu Agricultural and Ecological Experimental Station, Shaanxi, we evaluated the responses of soil microbial activity and crop to various management systems. The treatments were NMC (no mulching with inorganic N fertilizer, GMC (gravel mulching with inorganic N fertilizer, FMC (plastic-film mulching with inorganic N fertilizer and FMO (plastic-film mulching with inorganic N fertilizer and organic manure addition. The results showed that the FMO soil had the highest contents of microbial biomass carbon and nitrogen, dehydrogenase activity, microbial activity and Shannon diversity index. The relative use of carbohydrates and amino acids by microbes was highest in the FMO soil, whereas the relative use of polymers, phenolic compounds and amines was highest in the soil in the NMC soil. Compared with the NMC, an increased but no significant trend of biomass production and nitrogen accumulation was observed under the GMC treatment. The FMC and FMO led a greater increase in biomass production than GMC and NMC. Compare with the NMC treatment, FMC increased grain yield, maize biomass and nitrogen accumulation by 62.2, 62.9 and 86.2%, but no significant difference was found between the FMO and FMC treatments. Some soil biological properties, i.e. microbial biomass carbon, microbial biomass nitrogen, being sensitive to the mulching and organic fertilizer, were significant correlated with yield and nitrogen availability. Film mulching over gravel mulching can serve as an effective measure for crop production and nutrient cycling, and plus organic fertilization additions may thus have improvements in the biological

  3. A Broader View: Microbial Enzymes and Their Relevance in Industries, Medicine, and Beyond

    Science.gov (United States)

    Bose, Sutapa; Rai, Vivek

    2013-01-01

    Enzymes are the large biomolecules that are required for the numerous chemical interconversions that sustain life. They accelerate all the metabolic processes in the body and carry out a specific task. Enzymes are highly efficient, which can increase reaction rates by 100 million to 10 billion times faster than any normal chemical reaction. Due to development in recombinant technology and protein engineering, enzymes have evolved as an important molecule that has been widely used in different industrial and therapeutical purposes. Microbial enzymes are currently acquiring much attention with rapid development of enzyme technology. Microbial enzymes are preferred due to their economic feasibility, high yields, consistency, ease of product modification and optimization, regular supply due to absence of seasonal fluctuations, rapid growth of microbes on inexpensive media, stability, and greater catalytic activity. Microbial enzymes play a major role in the diagnosis, treatment, biochemical investigation, and monitoring of various dreaded diseases. Amylase and lipase are two very important enzymes that have been vastly studied and have great importance in different industries and therapeutic industry. In this review, an approach has been made to highlight the importance of different enzymes with special emphasis on amylase and lipase in the different industrial and medical fields. PMID:24106701

  4. Microbial Decomposers Not Constrained by Climate History Along a Mediterranean Climate Gradient

    Science.gov (United States)

    Baker, N. R.; Khalili, B.; Martiny, J. B. H.; Allison, S. D.

    2017-12-01

    The return of organic carbon to the atmosphere through terrestrial decomposition is mediated through the breakdown of complex organic polymers by extracellular enzymes produced by microbial decomposer communities. Determining if and how these decomposer communities are constrained in their ability to degrade plant litter is necessary for predicting how carbon cycling will be affected by future climate change. To address this question, we deployed fine-pore nylon mesh "microbial cage" litterbags containing grassland litter with and without local inoculum across five sites in southern California, spanning a gradient of 10.3-22.8° C in mean annual temperature and 100-400+ mm mean annual precipitation. Litterbags were deployed in October 2014 and collected four times over the course of 14 months. Recovered litter was assayed for mass loss, litter chemistry, microbial biomass, extracellular enzymes (Vmax and Km­), and enzyme temperature sensitivities. We hypothesized that grassland litter would decompose most rapidly in the grassland site, and that access to local microbial communities would enhance litter decomposition rates and microbial activity in the other sites along the gradient. We determined that temperature and precipitation likely interact to limit microbial decomposition in the extreme sites along our gradient. Despite their unique climate history, grassland microbes were not restricted in their ability to decompose litter under different climate conditions. Although we observed a strong correlation between bacterial biomass and mass loss across the gradient, litter that was inoculated with local microbial communities lost less mass despite having greater bacterial biomass and potentially accumulating more microbial residues. Our results suggest that microbial community composition may not constrain C-cycling rates under climate change in our system. However, there may be community constraints on decomposition if climate change alters litter chemistry, a

  5. Application of next-generation sequencing methods for microbial monitoring of anaerobic digestion of lignocellulosic biomass.

    Science.gov (United States)

    Bozan, Mahir; Akyol, Çağrı; Ince, Orhan; Aydin, Sevcan; Ince, Bahar

    2017-09-01

    The anaerobic digestion of lignocellulosic wastes is considered an efficient method for managing the world's energy shortages and resolving contemporary environmental problems. However, the recalcitrance of lignocellulosic biomass represents a barrier to maximizing biogas production. The purpose of this review is to examine the extent to which sequencing methods can be employed to monitor such biofuel conversion processes. From a microbial perspective, we present a detailed insight into anaerobic digesters that utilize lignocellulosic biomass and discuss some benefits and disadvantages associated with the microbial sequencing techniques that are typically applied. We further evaluate the extent to which a hybrid approach incorporating a variation of existing methods can be utilized to develop a more in-depth understanding of microbial communities. It is hoped that this deeper knowledge will enhance the reliability and extent of research findings with the end objective of improving the stability of anaerobic digesters that manage lignocellulosic biomass.

  6. A Compilation of Global Soil Microbial Biomass Carbon, Nitrogen, and Phosphorus Data

    Data.gov (United States)

    National Aeronautics and Space Administration — This data set provides the concentrations of soil microbial biomass carbon (C), nitrogen (N) and phosphorus (P), soil organic carbon, total nitrogen, and total...

  7. Early-branching Gut Fungi Possess A Large, And Comprehensive Array Of Biomass-Degrading Enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Solomon, Kevin V.; Haitjema, Charles; Henske, John K.; Gilmore, Sean P.; Borges-Rivera, Diego; Lipzen, Anna; Brewer, Heather M.; Purvine, Samuel O.; Wright, Aaron T.; Theodorou, Michael K.; Grigoriev, Igor V.; Regev, Aviv; Thompson, Dawn; O' Malley, Michelle A.

    2016-03-11

    The fungal kingdom is the source of almost all industrial enzymes in use for lignocellulose bioprocessing. Its more primitive members, however, remain relatively unexploited. We developed a systems-level approach that integrates RNA-Seq, proteomics, phenotype and biochemical studies of relatively unexplored early-branching free-living fungi. Anaerobic gut fungi isolated from herbivores produce a large array of biomass-degrading enzymes that synergistically degrade crude, unpretreated plant biomass, and are competitive with optimized commercial preparations from Aspergillus and Trichoderma. Compared to these model platforms, gut fungal enzymes are unbiased in substrate preference due to a wealth of xylan-degrading enzymes. These enzymes are universally catabolite repressed, and are further regulated by a rich landscape of noncoding regulatory RNAs. Furthermore, we identified several promising sequence divergent enzyme candidates for lignocellulosic bioprocessing.

  8. Farm Deployable Microbial Bioreactor for Fuel Ethanol Production

    Energy Technology Data Exchange (ETDEWEB)

    Okeke, Benedict [Auburn Univ., Montgomery AL (United States)

    2016-03-30

    Research was conducted to develop a farm and field deployable microbial bioreactor for bioethanol production from biomass. Experiments were conducted to select the most efficient microorganisms for conversion of plant fiber to sugars for fermentation to ethanol. Mixtures of biomass and surface soil samples were collected from selected sites in Alabama black belt counties (Macon, Sumter, Choctaw, Dallas, Montgomery, Lowndes) and other areas within the state of Alabama. Experiments were conducted to determine the effects of culture parameters on key biomass saccharifying enzymes (cellulase, beta-glucosidase, xylanase and beta-xylosidase). A wide-scale sampling of locally-grown fruits in Central Alabama was embarked to isolate potential xylose fermenting microorganisms. Yeast isolates were evaluated for xylose fermentation. Selected microorganisms were characterized by DNA based methods. Factors affecting enzyme production and biomass saccharification were examined and optimized in the laboratory. Methods of biomass pretreatment were compared. Co-production of amylolytic enzymes with celluloytic-xylanolytic enzymes was evaluated; and co-saccharification of a combination of biomass, and starch-rich materials was examined. Simultaneous saccharification and fermentation with and without pre-saccharifcation was studied. Whole culture broth and filtered culture broth simultaneous saccahrifcation and fermentation were compared. A bioreactor system was designed and constructed to employ laboratory results for scale up of biomass saccharification.

  9. Microbial enzyme activities of peatland soils in south central Alaska lowlands

    Science.gov (United States)

    Microbial enzyme activities related to carbon and nutrient acquisition were measured on Alaskan peatland soils as indicators of nutrient limitation and biochemical sustainability. Peat decomposition is mediated by microorganisms and enzymes that in turn are limited by various ph...

  10. Effects of 1-Alkyl-3-Methylimidazolium Nitrate on Soil Physical and Chemical Properties and Microbial Biomass.

    Science.gov (United States)

    Zhou, Tongtong; Wang, Jun; Ma, Zhiqiang; Du, Zhongkun; Zhang, Cheng; Zhu, Lusheng; Wang, Jinhua

    2018-05-01

    Ionic liquids (ILs), also called room temperature ILs, are widely applied in many fields on the basis of their unique physical and chemical properties. However, numerous ILs may be released into and gradually accumulate in the environment due to their extensive use and absolute solubility. The effects of 1-alkyl-3-methylimidazolium nitrate ([C n mim]NO 3 , n = 4, 6, 8) on soil pH, conductivity, cation exchange capacity, microbial biomass carbon, and microbial biomass nitrogen were examined at the doses of 1, 10, and 100 mg/kg on days 10, 20, 30, and 40. The results demonstrated that the soil pH decreased and the conductivity increased with increasing IL doses. No significant differences were observed in the soil cation-exchange capacity. All three of the tested ILs decreased the soil microbial biomass carbon and nitrogen. Additionally, there were few differences among the ILs with different alkyl chain lengths on the tested indicators except for the microbial biomass nitrogen. The present study addressed a gap in the literature regarding the effects of the aforementioned ILs with different alkyl side chains on the physicochemical properties of soil, and the results could provide the basic data for future studies on their toxicity to soil organisms, such as earthworms and soil microbes.

  11. Soil microbial biomass under different management and tillage systems of permanent intercropped cover species in an orange orchard

    Directory of Open Access Journals (Sweden)

    Elcio Liborio Balota

    2011-12-01

    Full Text Available To mitigate soil erosion and enhance soil fertility in orange plantations, the permanent protection of the inter-rows by cover species has been suggested. The objective of this study was to evaluate alterations in the microbial biomass, due to different soil tillage systems and intercropped cover species between rows of orange trees. The soil of the experimental area previously used as pasture (Brachiaria humidicola was an Ultisol (Typic Paleudult originating from Caiuá sandstone in the northwestern part of the State of Paraná, Brazil. Two soil tillage systems were evaluated: conventional tillage (CT in the entire area and strip tillage (ST (strip width 2 m, in combination with different ground cover management systems. The citrus cultivar 'Pera' orange (Citrus sinensis grafted onto 'Rangpur' lime rootstock was used. Soil samples were collected after five years of treatment from a depth of 0-15 cm, under the tree canopy and in the inter-row, in the following treatments: (1 CT and an annual cover crop with the leguminous species Calopogonium mucunoides; (2 CT and a perennial cover crop with the leguminous peanut Arachis pintoi; (3 CT and an evergreen cover crop with Bahiagrass Paspalum notatum; (4 CT and a cover crop with spontaneous Brachiaria humidicola grass vegetation; and (5 ST and maintenance of the remaining grass (pasture of Brachiaria humidicola. Soil tillage and the different cover species influenced the microbial biomass, both under the tree canopy and in the inter-row. The cultivation of brachiaria increased C and N in the microbial biomass, while bahiagrass increased P in the microbial biomass. The soil microbial biomass was enriched in N and P by the presence of ground cover species and according to the soil P content. The grass species increased C, N and P in the soil microbial biomass from the inter-row more than leguminous species.

  12. High-solids enrichment of thermophilic microbial communities and their enzymes on bioenergy feedstocks

    Energy Technology Data Exchange (ETDEWEB)

    Reddy, A. P.; Allgaier, M.; Singer, S.W.; Hazen, T.C.; Simmons, B.A.; Hugenholtz, P.; VanderGheynst, J.S.

    2011-04-01

    Thermophilic microbial communities that are active in a high-solids environment offer great potential for the discovery of industrially relevant enzymes that efficiently deconstruct bioenergy feedstocks. In this study, finished green waste compost was used as an inoculum source to enrich microbial communities and associated enzymes that hydrolyze cellulose and hemicellulose during thermophilic high-solids fermentation of the bioenergy feedstocks switchgrass and corn stover. Methods involving the disruption of enzyme and plant cell wall polysaccharide interactions were developed to recover xylanase and endoglucanase activity from deconstructed solids. Xylanase and endoglucanase activity increased by more than a factor of 5, upon four successive enrichments on switchgrass. Overall, the changes for switchgrass were more pronounced than for corn stover; solids reduction between the first and second enrichments increased by a factor of four for switchgrass while solids reduction remained relatively constant for corn stover. Amplicon pyrosequencing analysis of small-subunit ribosomal RNA genes recovered from enriched samples indicated rapid changes in the microbial communities between the first and second enrichment with the simplified communities achieved by the third enrichment. The results demonstrate a successful approach for enrichment of unique microbial communities and enzymes active in a thermophilic high-solids environment.

  13. The ecological effects of different loading rates of metalaxyl on microbial biomass in unplanted and planted soils under field conditions

    Directory of Open Access Journals (Sweden)

    M. Mansourzadeh

    2016-05-01

    Full Text Available Fungicides are most widely used pesticides in Iran and the world. Application of fungicides may affect the populations and activity of soil microorganisms, particularly fungi, with a consequence for soil fertility and crop growth. In the current study, the effects of different levels of metalaxyl on soil microbial biomass carbon (C and nitrogen (N, microbial biomass C/N ratio and metabolic quotient under field conditions were assessed. Two levels of metalaxyl (30 and 60 kg.ha-1 were applied in planted soils with corn and unplanted calcareous soils, using a split-plots experiment in a completely randomized design with three replications. The C and N contents in soil microbial biomass as well as metabolic quotient were measured at 30 and 90 days after the onset of the experiment. Results showed that in cultivated soils metalaxyl application at 30 kg.ha-1 increased (15-80% significantly (p≤0.01 the amounts of microbial biomass C and N at both intervals (except microbial biomass C at 90 days compared to the control soil (0 kg.ha-1, while in uncultivated soils both microbial biomass C and N reduced by almost 1-34%. Microbial biomass C/N ratios in unplanted soils decreased (15 and 53% with increasing loading rates of metalaxyl, without a clear effect in cultivated soils. On the other hand, metabolic quotient values reduced (48% at 30 and 60 kg.ha-1 metalaxyl in corn-cultivated soils when compared to untreated soils while in uncultivated soils metalaxyl rate at 30 kg.a-1 had the greatest values at 30 days, and increased with increasing the levels of metalaxyl at 90 days. In summary, application of metalaxyl can either reduce or increase soil biological indices, and the direction and changes are depended upon the application rate of metalaxyl, time elapsed since metalaxyl application and the presence or absence of plant.

  14. Soil respiration, microbial biomass and exoenzyme activity in switchgrass stands under nitrogen fertilization management and climate warming.

    Science.gov (United States)

    Jian, S.; Li, J.; de Koff, J.; Celada, S.; Mayes, M. A.; Wang, G.; Guo, C.

    2016-12-01

    Switchgrass (Panicum virgatum L.), as a model bioenergy crop, received nitrogen fertilizers for increasing its biomass yields. Studies rarely investigate the interactive effects of nitrogen fertilization and climate warming on soil microbial activity and carbon cycling in switchgrass cropping systems. Enhanced nitrogen availability under fertilization can alter rates of soil organic matter decomposition and soil carbon emissions to the atmosphere and thus have an effect on climate change. Here, we assess soil CO2 emission, microbial biomass and exoenzyme activities in two switchgrass stands with no fertilizer and 60 lbs N / acre. Soils were incubated at 15 ºC and 20 ºC for 180-day. Dry switchgrass plant materials were added to incubation jars and the 13C stable isotopic probing technique was used to monitor soil CO2 respiration derived from relatively labile litter and indigenous soil. Measurements of respiration, δ13C of respiration, microbial biomass carbon and exoenzyme activity were performed on days 1, 5, 10, 15, 30, 60, 90, 120, 150 and 180. Soil respiration rate was greater in the samples incubated at 20 ºC as compared to those incubated at 15 ºC. Exoenzyme activities were significantly altered by warming, litter addition and nitrogen fertilization. There was a significant interactive effect of nitrogen fertilization and warming on the proportion of CO2 respired from soils such that nitrogen fertilization enhanced warming-induced increase by 12.0% (Pmineralization. Fertilization increased soil microbial biomass carbon at both temperatures (9.0% at 15 ºC and 14.5% at 20 ºC). Our preliminary analysis suggested that warming effects on enhanced soil respiration can be further increased with elevated fertilizer input via greater microbial biomass and exoenzyme activity. In addition to greater biomass yield under N fertilization, this study informs potential soil carbon loss from stimulated soil respiration under nitrogen fertilization and warming in

  15. [Characteristics of soil microbes and enzyme activities in different degraded alpine meadows].

    Science.gov (United States)

    Yin, Ya Li; Wang, Yu Qin; Bao, Gen Sheng; Wang, Hong Sheng; Li, Shi Xiong; Song, Mei Ling; Shao, Bao Lian; Wen, Yu Cun

    2017-12-01

    Soil microbial biomass C and N, microbial diversities and enzyme activity in 0-10 cm and 10-20 cm soil layers of different degraded grasslands (non-degradation, ND; light degradation, LD; moderate degradation, MD; sever degradation, SD; and black soil beach, ED) were measured by Biolog and other methods. The results showed that: 1) There were significant diffe-rences between 0-10 cm and 10-20 cm soil layers in soil microbial biomass, diversities and inver-tase activities in all grasslands. 2) The ratio of soil microbial biomass C to N decreased significantly with the grassland degradation. In the 0-10 cm soil layer, microbial biomass C and N in ND and LD were significantly higher than that in MD, SD and ED. Among the latter three kinds of grasslands, there was no difference for microbial biomass C, but microbial biomass N was lower in MD than in the other grasslands. The average color change rate (AWCD) and McIntosh Index (U) also decreased with grassland degradation, but only the reduction from ND to MD was significant. There were no differences among all grasslands for Shannon index (H) and Simpson Index (D). The urease activity was highest in MD and SD, and the activity of phosphatase and invertase was lowest in ED. In the 10-20 cm soil layer, microbial biomass C in ND and LD were significantly higher than that in the other grasslands. Microbial biomass N in LD and ED were significantly higher than that in the other grasslands. Carbon metabolism index in MD was significantly lower than that in LD and SD. AWCD and U index in ND and LD were significantly higher than that in ED. H index and D index showed no difference among different grasslands. The urease activity in ND and MD was significantly higher than that in the other grasslands. The phosphatase activity was highest in MD, and the invertase activity was lowest in MD. 3) The belowground biomass was significantly positively correlated with microbial biomass, carbon metabolic index and phosphatase activity

  16. Effects of Biochar on Soil Microbial Biomass after Four Years of Consecutive Application in the North China Plain

    Science.gov (United States)

    Zhang, Qing-zhong; Dijkstra, Feike A.; Liu, Xing-ren; Wang, Yi-ding; Huang, Jian; Lu, Ning

    2014-01-01

    The long term effect of biochar application on soil microbial biomass is not well understood. We measured soil microbial biomass carbon (MBC) and nitrogen (MBN) in a field experiment during a winter wheat growing season after four consecutive years of no (CK), 4.5 (B4.5) and 9.0 t biochar ha−1 yr−1 (B9.0) applied. For comparison, a treatment with wheat straw residue incorporation (SR) was also included. Results showed that biochar application increased soil MBC significantly compared to the CK treatment, and that the effect size increased with biochar application rate. The B9.0 treatment showed the same effect on MBC as the SR treatment. Treatments effects on soil MBN were less strong than for MBC. The microbial biomass C∶N ratio was significantly increased by biochar. Biochar might decrease the fraction of biomass N mineralized (K N), which would make the soil MBN for biochar treatments underestimated, and microbial biomass C∶N ratios overestimated. Seasonal fluctuation in MBC was less for biochar amended soils than for CK and SR treatments, suggesting that biochar induced a less extreme environment for microorganisms throughout the season. There was a significant positive correlation between MBC and soil water content (SWC), but there was no significant correlation between MBC and soil temperature. Biochar amendments may therefore reduce temporal variability in environmental conditions for microbial growth in this system thereby reducing temporal fluctuations in C and N dynamics. PMID:25025330

  17. Effects of biochar on soil microbial biomass after four years of consecutive application in the north China Plain.

    Directory of Open Access Journals (Sweden)

    Qing-zhong Zhang

    Full Text Available The long term effect of biochar application on soil microbial biomass is not well understood. We measured soil microbial biomass carbon (MBC and nitrogen (MBN in a field experiment during a winter wheat growing season after four consecutive years of no (CK, 4.5 (B4.5 and 9.0 t biochar ha(-1 yr(-1 (B9.0 applied. For comparison, a treatment with wheat straw residue incorporation (SR was also included. Results showed that biochar application increased soil MBC significantly compared to the CK treatment, and that the effect size increased with biochar application rate. The B9.0 treatment showed the same effect on MBC as the SR treatment. Treatments effects on soil MBN were less strong than for MBC. The microbial biomass C∶N ratio was significantly increased by biochar. Biochar might decrease the fraction of biomass N mineralized (KN, which would make the soil MBN for biochar treatments underestimated, and microbial biomass C∶N ratios overestimated. Seasonal fluctuation in MBC was less for biochar amended soils than for CK and SR treatments, suggesting that biochar induced a less extreme environment for microorganisms throughout the season. There was a significant positive correlation between MBC and soil water content (SWC, but there was no significant correlation between MBC and soil temperature. Biochar amendments may therefore reduce temporal variability in environmental conditions for microbial growth in this system thereby reducing temporal fluctuations in C and N dynamics.

  18. Integrative computational approach for genome-based study of microbial lipid-degrading enzymes.

    Science.gov (United States)

    Vorapreeda, Tayvich; Thammarongtham, Chinae; Laoteng, Kobkul

    2016-07-01

    Lipid-degrading or lipolytic enzymes have gained enormous attention in academic and industrial sectors. Several efforts are underway to discover new lipase enzymes from a variety of microorganisms with particular catalytic properties to be used for extensive applications. In addition, various tools and strategies have been implemented to unravel the functional relevance of the versatile lipid-degrading enzymes for special purposes. This review highlights the study of microbial lipid-degrading enzymes through an integrative computational approach. The identification of putative lipase genes from microbial genomes and metagenomic libraries using homology-based mining is discussed, with an emphasis on sequence analysis of conserved motifs and enzyme topology. Molecular modelling of three-dimensional structure on the basis of sequence similarity is shown to be a potential approach for exploring the structural and functional relationships of candidate lipase enzymes. The perspectives on a discriminative framework of cutting-edge tools and technologies, including bioinformatics, computational biology, functional genomics and functional proteomics, intended to facilitate rapid progress in understanding lipolysis mechanism and to discover novel lipid-degrading enzymes of microorganisms are discussed.

  19. Soil microbial biomass, activity and community composition along altitudinal gradients in the High Arctic (Billefjorden, Svalbard)

    Science.gov (United States)

    Kotas, Petr; Šantrůčková, Hana; Elster, Josef; Kaštovská, Eva

    2018-03-01

    The unique and fragile High Arctic ecosystems are vulnerable to global climate warming. The elucidation of factors driving microbial distribution and activity in arctic soils is essential for a comprehensive understanding of ecosystem functioning and its response to environmental change. The goals of this study were to investigate microbial biomass and activity, microbial community structure (MCS), and their environmental controls in soils along three elevational transects in the coastal mountains of Billefjorden, central Svalbard. Soils from four different altitudes (25, 275, 525 and 765 m above sea level) were analyzed for a suite of characteristics including temperature regimes, organic matter content, base cation availability, moisture, pH, potential respiration, and microbial biomass and community structure using phospholipid fatty acids (PLFAs). We observed significant spatial heterogeneity of edaphic properties among transects, resulting in transect-specific effects of altitude on most soil parameters. We did not observe any clear elevation pattern in microbial biomass, and microbial activity revealed contrasting elevational patterns between transects. We found relatively large horizontal variability in MCS (i.e., between sites of corresponding elevation in different transects), mainly due to differences in the composition of bacterial PLFAs, but also a systematic altitudinal shift in MCS related to different habitat preferences of fungi and bacteria, which resulted in high fungi-to-bacteria ratios at the most elevated sites. The biological soil crusts on these most elevated, unvegetated sites can host microbial assemblages of a size and activity comparable to those of the arctic tundra ecosystem. The key environmental factors determining horizontal and vertical changes in soil microbial properties were soil pH, organic carbon content, soil moisture and Mg2+ availability.

  20. Soil microbial biomass, activity and community composition along altitudinal gradients in the High Arctic (Billefjorden, Svalbard

    Directory of Open Access Journals (Sweden)

    P. Kotas

    2018-03-01

    Full Text Available The unique and fragile High Arctic ecosystems are vulnerable to global climate warming. The elucidation of factors driving microbial distribution and activity in arctic soils is essential for a comprehensive understanding of ecosystem functioning and its response to environmental change. The goals of this study were to investigate microbial biomass and activity, microbial community structure (MCS, and their environmental controls in soils along three elevational transects in the coastal mountains of Billefjorden, central Svalbard. Soils from four different altitudes (25, 275, 525 and 765 m above sea level were analyzed for a suite of characteristics including temperature regimes, organic matter content, base cation availability, moisture, pH, potential respiration, and microbial biomass and community structure using phospholipid fatty acids (PLFAs. We observed significant spatial heterogeneity of edaphic properties among transects, resulting in transect-specific effects of altitude on most soil parameters. We did not observe any clear elevation pattern in microbial biomass, and microbial activity revealed contrasting elevational patterns between transects. We found relatively large horizontal variability in MCS (i.e., between sites of corresponding elevation in different transects, mainly due to differences in the composition of bacterial PLFAs, but also a systematic altitudinal shift in MCS related to different habitat preferences of fungi and bacteria, which resulted in high fungi-to-bacteria ratios at the most elevated sites. The biological soil crusts on these most elevated, unvegetated sites can host microbial assemblages of a size and activity comparable to those of the arctic tundra ecosystem. The key environmental factors determining horizontal and vertical changes in soil microbial properties were soil pH, organic carbon content, soil moisture and Mg2+ availability.

  1. Bioaugmentation for Electricity Generation from Corn Stover Biomass Using Microbial Fuel Cells

    KAUST Repository

    Wang, Xin; Feng, Yujie; Wang, Heming; Qu, Youpeng; Yu, Yanling; Ren, Nanqi; Li, Nan; Wang, Elle; Lee, He; Logan, Bruce E.

    2009-01-01

    of microbial consortia specifically acclimated for biomass breakdown. A mixed culture that was developed to have a high saccharification rate with corn stover was added to singlechamber, air-cathode MFCs acclimated for power production using glucose. The MFC

  2. Effects of Hurricane-Felled Tree Trunks on Soil Carbon, Nitrogen, Microbial Biomass, and Root Length in a Wet Tropical Forest

    Directory of Open Access Journals (Sweden)

    D. Jean Lodge

    2016-11-01

    Full Text Available Decaying coarse woody debris can affect the underlying soil either by augmenting nutrients that can be exploited by tree roots, or by diminishing nutrient availability through stimulation of microbial nutrient immobilization. We analyzed C, N, microbial biomass C and root length in closely paired soil samples taken under versus 20–50 cm away from large trunks of two species felled by Hugo (1989 and Georges (1998 three times during wet and dry seasons over the two years following the study conducted by Georges. Soil microbial biomass, % C and % N were significantly higher under than away from logs felled by both hurricanes (i.e., 1989 and 1998, at all sampling times and at both depths (0–10 and 10–20 cm. Frass from wood boring beetles may contribute to early effects. Root length was greater away from logs during the dry season, and under logs in the wet season. Root length was correlated with microbial biomass C, soil N and soil moisture (R = 0.36, 0.18, and 0.27, respectively; all p values < 0.05. Microbial biomass C varied significantly among seasons but differences between positions (under vs. away were only suggestive. Microbial C was correlated with soil N (R = 0.35. Surface soil on the upslope side of the logs had significantly more N and microbial biomass, likely from accumulation of leaf litter above the logs on steep slopes. We conclude that decaying wood can provide ephemeral resources that are exploited by tree roots during some seasons.

  3. Colloid-based multiplexed method for screening plant biomass-degrading glycoside hydrolase activities in microbial communities

    Energy Technology Data Exchange (ETDEWEB)

    Reindl, W.; Deng, K.; Gladden, J.M.; Cheng, G.; Wong, A.; Singer, S.W.; Singh, S.; Lee, J.-C.; Yao, J.-S.; Hazen, T.C.; Singh, A.K; Simmons, B.A.; Adams, P.D.; Northen, T.R.

    2011-05-01

    The enzymatic hydrolysis of long-chain polysaccharides is a crucial step in the conversion of biomass to lignocellulosic biofuels. The identification and characterization of optimal glycoside hydrolases is dependent on enzyme activity assays, however existing methods are limited in terms of compatibility with a broad range of reaction conditions, sample complexity, and especially multiplexity. The method we present is a multiplexed approach based on Nanostructure-Initiator Mass Spectrometry (NIMS) that allowed studying several glycolytic activities in parallel under diverse assay conditions. Although the substrate analogs carried a highly hydrophobic perfluorinated tag, assays could be performed in aqueous solutions due colloid formation of the substrate molecules. We first validated our method by analyzing known {beta}-glucosidase and {beta}-xylosidase activities in single and parallel assay setups, followed by the identification and characterization of yet unknown glycoside hydrolase activities in microbial communities.

  4. Assessment of Soil Health in Urban Agriculture: Soil Enzymes and Microbial Properties

    Directory of Open Access Journals (Sweden)

    Avanthi Deshani Igalavithana

    2017-02-01

    Full Text Available Urban agriculture has been recently highlighted with the increased importance for recreation in modern society; however, soil quality and public health may not be guaranteed because of continuous exposure to various pollutants. The objective of this study was to evaluate the soil quality of urban agriculture by soil microbial assessments. Two independent variables, organic and inorganic fertilizers, were considered. The activities of soil enzymes including dehydrogenase, β-glucosidase, arylsulfatase, urease, alkaline and acid phosphatases were used as indicators of important microbial mediated functions and the soil chemical properties were measured in the soils applied with organic or inorganic fertilizer for 10 years. Fatty acid methyl ester analysis was applied to determine the soil microbial community composition. Relatively higher microbial community richness and enzyme activities were found in the organic fertilizers applied soils as compared to the inorganic fertilizers applied soils. Principal component analysis explained the positive influence of organic fertilizers on the microbial community. The application of organic fertilizers can be a better alternative compared to inorganic fertilizers for the long-term health and security of urban agriculture.

  5. Recombinant protein production facility for fungal biomass-degrading enzymes using the yeast Pichia pastoris

    Directory of Open Access Journals (Sweden)

    Mireille eHaon

    2015-09-01

    Full Text Available Filamentous fungi are the predominant source of lignocellulolytic enzymes used in industry for the transformation of plant biomass into high-value molecules and biofuels. The rapidity with which new fungal genomic and post-genomic data are being produced is vastly outpacing functional studies. This underscores the critical need for developing platforms dedicated to the recombinant expression of enzymes lacking confident functional annotation, a prerequisite to their functional and structural study. In the last decade, the yeast Pichia pastoris has become increasingly popular as a host for the production of fungal biomass-degrading enzymes, and particularly carbohydrate-active enzymes (CAZymes. This study aimed at setting-up a platform to easily and quickly screen the extracellular expression of biomass-degrading enzymes in Pichia pastoris. We first used three fungal glycoside hydrolases that we previously expressed using the protocol devised by Invitrogen to try different modifications of the original protocol. Considering the gain in time and convenience provided by the new protocol, we used it as basis to set-up the facility and produce a suite of fungal CAZymes (glycoside hydrolases, carbohydrate esterases and auxiliary activity enzyme families out of which more than 70% were successfully expressed. The platform tasks range from gene cloning to automated protein purifications and activity tests, and is open to the CAZyme users’ community.

  6. [Dynamics of microbial biomass carbon and nitrogen during foliar litter decomposition under artificial forest gap in Pinus massoniana plantation.

    Science.gov (United States)

    Zhang, Ming Jin; Chen, Liang Hua; Zhang, Jian; Yang, Wan Qin; Liu, Hua; Li, Xun; Zhang, Yan

    2016-03-01

    Nowadays large areas of plantations have caused serious ecological problems such as soil degradation and biodiversity decline. Artificial tending thinning and construction of mixed forest are frequently used ways when we manage plantations. To understand the effect of this operation mode on nutrient cycle of plantation ecosystem, we detected the dynamics of microbial bio-mass carbon and nitrogen during foliar litter decomposition of Pinus massoniana and Toona ciliate in seven types of gap in different sizes (G 1 : 100 m 2 , G 2 : 225 m 2 , G 3 : 400 m 2 , G 4 : 625 m 2 , G 5 : 900 m 2 , G 6 : 1225 m 2 , G 7 : 1600 m 2 ) of 42-year-old P. massoniana plantations in a hilly area of the upper Yang-tze River. The results showed that small and medium-sized forest gaps(G 1 -G 5 ) were more advantageous for the increment of microbial biomass carbon and nitrogen in the process of foliar litter decomposition. Along with the foliar litter decomposition during the experiment (360 d), microbial biomass carbon (MBC), microbial biomass nitrogen (MBN) in P. massoniana foliar litter and MBN in T. ciliata foliar litter first increased and then decreased, and respectively reached the maxima 9.87, 0.22 and 0.80 g·kg -1 on the 180 th d. But the peak (44.40 g·kg -1 ) of MBC in T. ciliata foliar litter appeared on the 90 th d. Microbial biomass carbon and nitrogen in T. ciliate was significantly higher than that of P. massoniana during foliar litter decomposition. Microbial biomass carbon and nitrogen in foliar litter was not only significantly associated with average daily temperature and the water content of foliar litter, but also closely related to the change of the quality of litter. Therefore, in the thinning, forest gap size could be controlled in the range of from 100 to 900 m 2 to facilitate the increase of microbial biomass carbon and nitrogen in the process of foliar litter decomposition, accelerate the decomposition of foliar litter and improve soil fertility of plantations.

  7. Extracellular enzyme activity assay as indicator of soil microbial functional diversity and activity

    DEFF Research Database (Denmark)

    Hendriksen, Niels Bohse; Winding, Anne

    2012-01-01

    Extracellular enzyme activity assay as indicator of soil microbial functional diversity and activity Niels Bohse Hendriksen, Anne Winding. Department of Environmental Science, Aarhus University, 4000 Roskilde, Denmark Soils provide numerous essential ecosystem services such as carbon cycling...... of soil microbial functions is still needed. In soil, enzymes originate from a variety of organisms, notably fungi and bacteria and especially hydrolytic extracellular enzymes are of pivotal importance for decomposition of organic substrates and biogeochemical cycling. Their activity will reflect...... the functional diversity and activity of the microorganisms involved in decomposition processes. Their activity has been measured by the use of fluorogenic model substrates e.g. methylumbelliferyl (MUF) substrates for a number of enzymes involved in the degradation of polysacharides as cellulose, hemicellulose...

  8. End-to-end gene fusions and their impact on the production of multifunctional biomass degrading enzymes

    International Nuclear Information System (INIS)

    Rizk, Mazen; Antranikian, Garabed; Elleuche, Skander

    2012-01-01

    Highlights: ► Multifunctional enzymes offer an interesting approach for biomass degradation. ► Size and conformation of separate constructs play a role in the effectiveness of chimeras. ► A connecting linker allows for maximal flexibility and increased thermostability. ► Genes with functional similarities are the best choice for fusion candidates. -- Abstract: The reduction of fossil fuels, coupled with its increase in price, has made the search for alternative energy resources more plausible. One of the topics gaining fast interest is the utilization of lignocellulose, the main component of plants. Its primary constituents, cellulose and hemicellulose, can be degraded by a series of enzymes present in microorganisms, into simple sugars, later used for bioethanol production. Thermophilic bacteria have proven to be an interesting source of enzymes required for hydrolysis since they can withstand high and denaturing temperatures, which are usually required for processes involving biomass degradation. However, the cost associated with the whole enzymatic process is staggering. A solution for cost effective and highly active production is through the construction of multifunctional enzyme complexes harboring the function of more than one enzyme needed for the hydrolysis process. There are various strategies for the degradation of complex biomass ranging from the regulation of the enzymes involved, to cellulosomes, and proteins harboring more than one enzymatic activity. In this review, the construction of multifunctional biomass degrading enzymes through end-to-end gene fusions, and its impact on production and activity by choosing the enzymes and linkers is assessed.

  9. The Physiology of Microbial Symbionts in Fungus-Farming Termites

    DEFF Research Database (Denmark)

    Rodrigues da Costa, Rafael

    . The termites provide the fungus with optimal growth conditions (e.g., stable temperature and humidity), as well as with constant inoculation of growth substrate and protection against alien fungi. In reward, the fungus provides the termites with a protein-rich fungal biomass based diet. In addition...... with their symbionts are main decomposer of organic matter in Africa, and this is reflect of a metabolic complementarity to decompose plant biomass in the genome of the three organisms involved in this symbiosis. Many of the physiological aspects of this symbiosis remain obscure, and here I focus on physiology...... of microbial symbionts associated with fungus-growing termites. Firstly, by using a set of enzyme assays, plant biomass compositional analyses, and RNA sequencing we gained deeper understanding on what enzymes are produced and active at different times of the decomposition process. Our results show that enzyme...

  10. The hemicellulolytic enzyme arsenal of Thermobacillus xylanilyticus depends on the composition of biomass used for growth

    Directory of Open Access Journals (Sweden)

    Rakotoarivonina Harivony

    2012-12-01

    Full Text Available Abstract Background Thermobacillus xylanilyticus is a thermophilic and highly xylanolytic bacterium. It produces robust and stable enzymes, including glycoside hydrolases and esterases, which are of special interest for the development of integrated biorefineries. To investigate the strategies used by T. xylanilyticus to fractionate plant cell walls, two agricultural by-products, wheat bran and straw (which differ in their chemical composition and tissue organization, were used in this study and compared with glucose and xylans. The ability of T. xylanilyticus to grow on these substrates was studied. When the bacteria used lignocellulosic biomass, the production of enzymes was evaluated and correlated with the initial composition of the biomass, as well as with the evolution of any residues during growth. Results Our results showed that T. xylanilyticus is not only able to use glucose and xylans as primary carbon sources but can also use wheat bran and straw. The chemical compositions of both lignocellulosic substrates were modified by T. xylanilyticus after growth. The bacteria were able to consume 49% and 20% of the total carbohydrates in bran and straw, respectively, after 24 h of growth. The phenolic and acetyl ester contents of these substrates were also altered. Bacterial growth on both lignocellulosic biomasses induced hemicellulolytic enzyme production, and xylanase was the primary enzyme secreted. Debranching activities were differentially produced, as esterase activities were more important to bacterial cultures grown on wheat straw; arabinofuranosidase production was significantly higher in bacterial cultures grown on wheat bran. Conclusion This study provides insight into the ability of T. xylanilyticus to grow on abundant agricultural by-products, which are inexpensive carbon sources for enzyme production. The composition of the biomass upon which the bacteria grew influenced their growth, and differences in the biomass provided

  11. Short- and long-term effects of nutrient enrichment on microbial exoenzyme activity in mangrove peat

    KAUST Repository

    Keuskamp, Joost A.

    2015-02-01

    © 2014 Elsevier Ltd. Mangroves receive increasing quantities of nutrients as a result of coastal development, which could lead to significant changes in carbon sequestration and soil subsidence. We hypothesised that mangrove-produced tannins induce a nitrogen (N) limitation on microbial decomposition even when plant growth is limited by phosphorus (P). As a result, increased N influx would lead to a net loss of sequestered carbon negating the ability to compensate for sea level rise in P-limited mangroves. To examine this, we quantified the short- and long-term effects of N and P enrichment on microbial biomass and decomposition-related enzyme activities in a Rhizophora mangle-dominated mangrove, which had been subjected to fertilisation treatments for a period of fifteen years. We compared microbial biomass, elemental stoichiometry and potential enzyme activity in dwarf and fringe-type R. mangle-dominated sites, where primary production is limited by P or N depending on the proximity to open water. Even in P-limited mangroves, microbial activity was N-limited as indicated by stoichiometry and an increase in enzymic activity upon N amendment. Nevertheless, microbial biomass increased upon field additions of P, indicating that the carbon supply played even a larger role. Furthermore, we found that P amendment suppressed phenol oxidase activity, while N amendment did not. The possible differential nutrient limitations of microbial decomposers versus primary producers implies that the direction of the effect of eutrophication on carbon sequestration is nutrient-specific. In addition, this study shows that phenol oxidase activities in this system decrease through P, possibly strengthening the enzymic latch effect of mangrove tannins. Furthermore, it is argued that the often used division between N-harvesting, P-harvesting, and carbon-harvesting exoenzymes needs to be reconsidered.

  12. Microbial respiration, but not biomass, responded linearly to increasing light fraction organic matter input: Consequences for carbon sequestration.

    Science.gov (United States)

    Rui, Yichao; Murphy, Daniel V; Wang, Xiaoli; Hoyle, Frances C

    2016-10-18

    Rebuilding 'lost' soil carbon (C) is a priority in mitigating climate change and underpinning key soil functions that support ecosystem services. Microorganisms determine if fresh C input is converted into stable soil organic matter (SOM) or lost as CO 2 . Here we quantified if microbial biomass and respiration responded positively to addition of light fraction organic matter (LFOM, representing recent inputs of plant residue) in an infertile semi-arid agricultural soil. Field trial soil with different historical plant residue inputs [soil C content: control (tilled) = 9.6 t C ha -1 versus tilled + plant residue treatment (tilled + OM) = 18.0 t C ha -1 ] were incubated in the laboratory with a gradient of LFOM equivalent to 0 to 3.8 t C ha -1 (0 to 500% LFOM). Microbial biomass C significantly declined under increased rates of LFOM addition while microbial respiration increased linearly, leading to a decrease in the microbial C use efficiency. We hypothesise this was due to insufficient nutrients to form new microbial biomass as LFOM input increased the ratio of C to nitrogen, phosphorus and sulphur of soil. Increased CO 2 efflux but constrained microbial growth in response to LFOM input demonstrated the difficulty for C storage in this environment.

  13. Effect of buctril super (Bromoxynil herbicide on soil microbial biomass and bacterial population

    Directory of Open Access Journals (Sweden)

    Zafar Abbas

    2014-02-01

    Full Text Available The present study aimed to evaluate the effect of bromoxynil herbicide on soil microorganisms, with the hypothesis that this herbicide caused suppression in microbial activity and biomass by exerting toxic effect on them. Nine sites of Punjab province (Pakistan those had been exposed to bromoxynil herbicide for about last ten years designated as soil 'A' were surveyed in 2011 and samples were collected and analyzed for Microbial Biomass Carbon (MBC, Biomass Nitrogen (MBN, Biomass Phosphorus (MBP and bacterial population. Simultaneously, soil samples from the same areas those were not exposed to herbicide designated as soil 'B' were taken. At all the sites MBC, MBN and MBP ranged from 131 to 457, 1.22 to 13.1 and 0.59 to 3.70 µg g-1 in the contaminated soils (Soil A, which was 187 to 573, 1.70 to 14.4 and 0.72 to 4.12 µg g-1 in the soils without contamination (soil B. Bacterial population ranged from 0.67 to 1.84x10(8 and 0.87 to 2.37x10(8 cfu g-1 soil in the soils A and B, respectively. Bromoxynil residues ranged from 0.09 to 0.24 mg kg-1 at all the sites in soil A. But no residues were detected in the soil B. Due to lethal effect of bromoxynil residues on the above parameters, considerable decline in these parameters was observed in the contaminated soils. Results depicted that the herbicide had left toxic effects on soil microbial parameters, thus confirmed that continuous use of this herbicide affected the quality of soil and sustainable crop production.

  14. Representing Microbial Dormancy in Soil Decomposition Models Improves Model Performance and Reveals Key Ecosystem Controls on Microbial Activity

    Science.gov (United States)

    He, Y.; Yang, J.; Zhuang, Q.; Wang, G.; Liu, Y.

    2014-12-01

    Climate feedbacks from soils can result from environmental change and subsequent responses of plant and microbial communities and nutrient cycling. Explicit consideration of microbial life history traits and strategy may be necessary to predict climate feedbacks due to microbial physiology and community changes and their associated effect on carbon cycling. In this study, we developed an explicit microbial-enzyme decomposition model and examined model performance with and without representation of dormancy at six temperate forest sites with observed soil efflux ranged from 4 to 10 years across different forest types. We then extrapolated the model to all temperate forests in the Northern Hemisphere (25-50°N) to investigate spatial controls on microbial and soil C dynamics. Both models captured the observed soil heterotrophic respiration (RH), yet no-dormancy model consistently exhibited large seasonal amplitude and overestimation in microbial biomass. Spatially, the total RH from temperate forests based on dormancy model amounts to 6.88PgC/yr, and 7.99PgC/yr based on no-dormancy model. However, no-dormancy model notably overestimated the ratio of microbial biomass to SOC. Spatial correlation analysis revealed key controls of soil C:N ratio on the active proportion of microbial biomass, whereas local dormancy is primarily controlled by soil moisture and temperature, indicating scale-dependent environmental and biotic controls on microbial and SOC dynamics. These developments should provide essential support to modeling future soil carbon dynamics and enhance the avenue for collaboration between empirical soil experiment and modeling in the sense that more microbial physiological measurements are needed to better constrain and evaluate the models.

  15. Microbial conversion technologies

    Energy Technology Data Exchange (ETDEWEB)

    Lau, P. [National Research Council of Canada, Ottawa, ON (Canada). Bioconversion and Sustainable Development

    2006-07-01

    Microbes are a biomass and an valuable resource. This presentation discussed microbial conversion technologies along with background information on microbial cells, their characteristics and microbial diversity. Untapped opportunities for microbial conversion were identified. Metagenomic and genome mining approaches were also discussed, as they can provide access to uncultivated or unculturable microorganisms in communal populations and are an unlimited resource for biocatalysts, novel genes and metabolites. Genome mining was seen as an economical approach. The presentation also emphasized that the development of microbial biorefineries would require significant insights into the relevant microorganisms and that biocatalysts were the ultimate in sustainability. In addition, the presentation discussed the natural fibres initiative for biochemicals and biomaterials. Anticipated outputs were identified and work in progress of a new enzyme-retting cocktail to provide diversity and/or consistency in fibre characteristics for various applications were also presented. It was concluded that it is necessary to leverage understanding of biological processes to produce bioproducts in a clean and sustainable manner. tabs., figs.

  16. Enzyme Amplified Detection of Microbial Cell Wall Components

    Science.gov (United States)

    Wainwright, Norman R.

    2004-01-01

    This proposal is MBL's portion of NASA's Johnson Space Center's Astrobiology Center led by Principal Investigator, Dr. David McKay, entitled: 'Institute for the Study of Biomarkers in Astromaterials.' Dr. Norman Wainwright is the principal investigator at MBL and is responsible for developing methods to detect trace quantities of microbial cell wall chemicals using the enzyme amplification system of Limulus polyphemus and other related methods.

  17. Potential and utilization of thermophiles and thermostable enzymes in biorefining

    Directory of Open Access Journals (Sweden)

    Karlsson Eva

    2007-03-01

    Full Text Available Abstract In today's world, there is an increasing trend towards the use of renewable, cheap and readily available biomass in the production of a wide variety of fine and bulk chemicals in different biorefineries. Biorefineries utilize the activities of microbial cells and their enzymes to convert biomass into target products. Many of these processes require enzymes which are operationally stable at high temperature thus allowing e.g. easy mixing, better substrate solubility, high mass transfer rate, and lowered risk of contamination. Thermophiles have often been proposed as sources of industrially relevant thermostable enzymes. Here we discuss existing and potential applications of thermophiles and thermostable enzymes with focus on conversion of carbohydrate containing raw materials. Their importance in biorefineries is explained using examples of lignocellulose and starch conversions to desired products. Strategies that enhance thermostablity of enzymes both in vivo and in vitro are also assessed. Moreover, this review deals with efforts made on developing vectors for expressing recombinant enzymes in thermophilic hosts.

  18. Rapid optimization of enzyme mixtures for deconstruction of diverse pretreatment/biomass feedstock combinations

    Directory of Open Access Journals (Sweden)

    Walton Jonathan D

    2010-10-01

    Full Text Available Abstract Background Enzymes for plant cell wall deconstruction are a major cost in the production of ethanol from lignocellulosic biomass. The goal of this research was to develop optimized synthetic mixtures of enzymes for multiple pretreatment/substrate combinations using our high-throughput biomass digestion platform, GENPLAT, which combines robotic liquid handling, statistical experimental design and automated Glc and Xyl assays. Proportions of six core fungal enzymes (CBH1, CBH2, EG1, β-glucosidase, a GH10 endo-β1,4-xylanase, and β-xylosidase were optimized at a fixed enzyme loading of 15 mg/g glucan for release of Glc and Xyl from all combinations of five biomass feedstocks (corn stover, switchgrass, Miscanthus, dried distillers' grains plus solubles [DDGS] and poplar subjected to three alkaline pretreatments (AFEX, dilute base [0.25% NaOH] and alkaline peroxide [AP]. A 16-component mixture comprising the core set plus 10 accessory enzymes was optimized for three pretreatment/substrate combinations. Results were compared to the performance of two commercial enzymes (Accellerase 1000 and Spezyme CP at the same protein loadings. Results When analyzed with GENPLAT, corn stover gave the highest yields of Glc with commercial enzymes and with the core set with all pretreatments, whereas corn stover, switchgrass and Miscanthus gave comparable Xyl yields. With commercial enzymes and with the core set, yields of Glc and Xyl were highest for grass stovers pretreated by AP compared to AFEX or dilute base. Corn stover, switchgrass and DDGS pretreated with AFEX and digested with the core set required a higher proportion of endo-β1,4-xylanase (EX3 and a lower proportion of endo-β1,4-glucanase (EG1 compared to the same materials pretreated with dilute base or AP. An optimized enzyme mixture containing 16 components (by addition of α-glucuronidase, a GH11 endoxylanase [EX2], Cel5A, Cel61A, Cip1, Cip2, β-mannanase, amyloglucosidase,

  19. A survey of Opportunities for Microbial Conversion of Biomass to Hydrocarbon Compatible Fuels

    Energy Technology Data Exchange (ETDEWEB)

    Jovanovic, Iva; Jones, Susanne B.; Santosa, Daniel M.; Dai, Ziyu; Ramasamy, Karthikeyan K.; Zhu, Yunhua

    2010-09-01

    Biomass is uniquely able to supply renewable and sustainable liquid transportation fuels. In the near term, the Biomass program has a 2012 goal of cost competitive cellulosic ethanol. However, beyond 2012, there will be an increasing need to provide liquid transportation fuels that are more compatible with the existing infrastructure and can supply fuel into all transportation sectors, including aviation and heavy road transport. Microbial organisms are capable of producing a wide variety of fuel and fuel precursors such as higher alcohols, ethers, esters, fatty acids, alkenes and alkanes. This report surveys liquid fuels and fuel precurors that can be produced from microbial processes, but are not yet ready for commercialization using cellulosic feedstocks. Organisms, current research and commercial activities, and economics are addressed. Significant improvements to yields and process intensification are needed to make these routes economic. Specifically, high productivity, titer and efficient conversion are the key factors for success.

  20. Soluble inhibitors/deactivators of cellulase enzymes from lignocellulosic biomass.

    Science.gov (United States)

    Kim, Youngmi; Ximenes, Eduardo; Mosier, Nathan S; Ladisch, Michael R

    2011-04-07

    Liquid hot water, steam explosion, and dilute acid pretreatments of lignocellulose generate soluble inhibitors which hamper enzymatic hydrolysis as well as fermentation of sugars to ethanol. Toxic and inhibitory compounds will vary with pretreatment and include soluble sugars, furan derivatives (hydroxymethyl fulfural, furfural), organic acids (acetic, formic and, levulinic acid), and phenolic compounds. Their effect is seen when an increase in the concentration of pretreated biomass in a hydrolysis slurry results in decreased cellulose conversion, even though the ratio of enzyme to cellulose is kept constant. We used lignin-free cellulose, Solka Floc, combined with mixtures of soluble components released during pretreatment of wood, to prove that the decrease in the rate and extent of cellulose hydrolysis is due to a combination of enzyme inhibition and deactivation. The causative agents were extracted from wood pretreatment liquid using PEG surfactant, activated charcoal or ethyl acetate and then desorbed, recovered, and added back to a mixture of enzyme and cellulose. At enzyme loadings of either 1 or 25mg protein/g glucan, the most inhibitory components, later identified as phenolics, decreased the rate and extent of cellulose hydrolysis by half due to both inhibition and precipitation of the enzymes. Full enzyme activity occurred when the phenols were removed. Hence detoxification of pretreated woods through phenol removal is expected to reduce enzyme loadings, and therefore reduce enzyme costs, for a given level of cellulose conversion. Copyright © 2011 Elsevier Inc. All rights reserved.

  1. Inorganic phosphorus fertilizer ameliorates maize growth by reducing metal uptake, improving soil enzyme activity and microbial community structure.

    Science.gov (United States)

    Wu, Wencheng; Wu, Jiahui; Liu, Xiaowen; Chen, Xianbin; Wu, Yingxin; Yu, Shixiao

    2017-09-01

    Recently, several studies have showed that both organic and inorganic fertilizers are effective in immobilizing heavy metals at low cost, in comparison to other remediation strategies for heavy metal-contaminated farmlands. A pot trial was conducted in this study to examine the effects of inorganic P fertilizer and organic fertilizer, in single application or in combination, on growth of maize, heavy metal availabilities, enzyme activities, and microbial community structure in metal-contaminated soils from an electronic waste recycling region. Results showed that biomass of maize shoot and root from the inorganic P fertilizer treatments were respectively 17.8 and 10.0 folds higher than the un-amended treatments (CK), while the biomass in the organic fertilizer treatments was only comparable to the CK. In addition, there were decreases of 85.0% in Cd, 74.3% in Pb, 66.3% in Cu, and 91.9% in Zn concentrations in the roots of maize grown in inorganic P fertilizer amended soil. Consistently, urease and catalase activities in the inorganic P fertilizer amended soil were 3.3 and 2.0 times higher than the CK, whereas no enhancement was observed in the organic fertilizer amended soil. Moreover, microbial community structure was improved by the application of inorganic P fertilizer, but not by organic fertilizer; the beneficial microbial groups such as Kaistobacter and Koribacter were most frequently detected in the inorganic P fertilizer amended soil. The negligible effect from the organic fertilizer might be ascribed to the decreased pH value in soils. The results suggest that the application of inorganic P fertilizer (or in combination with organic fertilizer) might be a promising strategy for the remediation of heavy metals contaminated soils in electronic waste recycling region. Copyright © 2017. Published by Elsevier Inc.

  2. Response of broiler chickens to diets containing artificially dried high-moisture maize supplemented with microbial enzymes

    OpenAIRE

    Bhuiyan, M.M; Islam, A.F; Iji, P.A

    2010-01-01

    The effect of feeding high-moisture maize grains dried in the sun or artificially in a forced draught oven at 80, 90 or 100 ºC for 24 hours and supplemented with microbial enzymes (Avizyme 1502 and Phyzyme XP) on growth performance, visceral organs, tissue protein, enzyme activity and gut development was investigated in a broiler growth trial. Feed intake (FI) up to 21 days decreased as a results of oven drying of grains whereas supplementation with microbial enzymes increased FI compared to ...

  3. Abundance and biomass responses of microbial food web components to hydrology and environmental gradients within a floodplain of the River Danube.

    Science.gov (United States)

    Palijan, Goran

    2012-07-01

    This study investigated the relationships of time-dependent hydrological variability and selected microbial food web components. Samples were collected monthly from the Kopački Rit floodplain in Croatia, over a period of 19 months, for analysis of bacterioplankton abundance, cell size and biomass; abundance of heterotrophic nanoflagellates and nanophytoplankton; and concentration of chlorophyll a. Similar hydrological variability at different times of the year enabled partition of seasonal effects from hydrological changes on microbial community properties. The results suggested that, unlike some other studies investigating sites with different connectivity, bacterioplankton abundance, and phytoplankton abundance and biomass increased during lentic conditions. At increasing water level, nanophytoplankton showed lower sensitivity to disturbance in comparison with total phytoplankton biomass: this could prolong autotrophic conditions within the floodplain. Bacterioplankton biomass, unlike phytoplankton, was not impacted by hydrology. The bacterial biomass less affected by hydrological changes can be an important additional food component for the floodplain food web. The results also suggested a mechanism controlling bacterial cell size independent of hydrology, as bacterial cell size was significantly decreased as nanoflagellate abundance increased. Hydrology, regardless of seasonal sucession, has the potential to structure microbial food webs, supporting microbial development during lentic conditions. Conversely, other components appear unaffected by hydrology or may be more strongly controlled by biotic interactions. This research, therefore, adds to understanding on microbial food web interactions in the context of flood and flow pulses in river-floodplain ecosystems.

  4. Microbial biomass and activity in litter during the initial development of pure and mixed plantations of Eucalyptus grandis and Acacia mangium

    Directory of Open Access Journals (Sweden)

    Daniel Bini

    2013-02-01

    Full Text Available Studies on microbial activity and biomass in forestry plantations often overlook the role of litter, typically focusing instead on soil nutrient contents to explain plant and microorganism development. However, since the litter is a significant source of recycled nutrients that affect nutrient dynamics in the soil, litter composition may be more strongly correlated with forest growth and development than soil nutrient contents. This study aimed to test this hypothesis by examining correlations between soil C, N, and P; litter C, N, P, lignin content, and polyphenol content; and microbial biomass and activity in pure and mixed second-rotation plantations of Eucalyptus grandis and Acacia mangium before and after senescent leaf drop. The numbers of cultivable fungi and bacteria were also estimated. All properties were correlated with litter C, N, P, lignin and polyphenols, and with soil C and N. We found higher microbial activity (CO2 evolution in litter than in soil. In the E. grandis monoculture before senescent leaf drop, microbial biomass C was 46 % higher in litter than in soil. After leaf drop, this difference decreased to 16 %. In A. mangium plantations, however, microbial biomass C was lower in litter than in soil both before and after leaf drop. Microbial biomass N of litter was approximately 94 % greater than that of the soil in summer and winter in all plantations. The number of cultivable fungi and bacteria increased after leaf drop, especially so in the litter. Fungi were also more abundant in the E. grandis litter. In general, the A. mangium monoculture was associated with higher levels of litter lignin and N, especially after leaf drop. In contrast, the polyphenol and C levels in E. grandis monoculture litter were higher after leaf drop. These properties were negatively correlated with total soil C and N. Litter in the mixed stands had lower C:N and C:P ratios and higher N, P, and C levels in the microbial biomass. This suggests more

  5. Winery biomass waste degradation by sequential sonication and mixed fungal enzyme treatments.

    Science.gov (United States)

    Karpe, Avinash V; Dhamale, Vijay V; Morrison, Paul D; Beale, David J; Harding, Ian H; Palombo, Enzo A

    2017-05-01

    To increase the efficiency of winery-derived biomass biodegradation, grape pomace was ultrasonicated for 20min in the presence of 0.25M, 0.5Mand1.0MKOH and 1.0MNaOH. This was followed by treatment with a 1:1 (v/v) mix of crude enzyme preparation derived from Phanerochaete chrysosporium and Trametes versicolor for 18h and a further 18h treatment with a 60:14:4:2 percent ratio combination of enzymes derived from Aspergillus niger: Penicillium chrysogenum: Trichoderma harzianum: P. citrinum, repsectively. Process efficiency was evaluated by its comparison to biological only mixed fungal degradation over 16days. Ultrasonication treatment with 0.5MKOH followed by mixed enzyme treatment yielded the highest lignin degradation of about 13%. Cellulase, β-glucosidase, xylanase, laccase and lignin peroxidase activities of 77.9, 476, 5,390.5, 66.7 and 29,230.7U/mL, respectively, were observed during biomass degradation. Gas chromatography-mass spectrometry (GC-MS) analysis of the degraded material identified commercially important compounds such as gallic acid, lithocholic acid, glycolic acid and lactic acid which were generated in considerable quantities. Thus, the combination of sonication pre-treatment and enzymatic degradation has the potential to considerably improve the breakdown of agricultural biomass and produce commercially useful compounds in markedly less time (<40h) with respect to biological only degradation (16days). Copyright © 2016 Elsevier Inc. All rights reserved.

  6. Temperature sensitivity differences with depth and season between carbon, nitrogen, and phosphorus cycling enzyme activities in an ombrotrophic peatland system

    Science.gov (United States)

    Steinweg, J. M.; Kostka, J. E.; Hanson, P. J.; Schadt, C. W.

    2017-12-01

    Northern peatlands have large amounts of soil organic matter due to reduced decomposition. Breakdown of organic matter is initially mediated by extracellular enzymes, the activity of which may be controlled by temperature, moisture, and substrate availability, all of which vary seasonally throughout the year and with depth. In typical soils the majority of the microbial biomass and decomposition occurs within the top 30cm due to reduced organic matter inputs in the subsurface however peatlands by their very nature contain large amounts of organic matter throughout their depth profile. We hypothesized that potential enzyme activity would be greatest at the surface of the peat due to a larger microbial biomass compared to 40cm and 175cm below the surface and that temperature sensitivity would be greatest at the surface during winter but lowest during the summer due to high temperatures and enzyme efficiency. Peat samples were collected in February, July, and August 2012 from the DOE Spruce and Peatland Responses Under Climatic and Environmental Change project at Marcell Experimental Forest S1 bog. We measured potential activity of hydrolytic enzymes involved in three different nutrient cycles: beta-glucosidase (carbon), leucine amino peptidase (nitrogen), and phosphatase (phosphorus) at 15 temperature points ranging from 3°C to 65°C. Enzyme activity decreased with depth as expected but there was no concurrent change in activation energy (Ea). The reduction in enzyme activity with depth indicates a smaller pool which coincided with a decreased microbial biomass. Differences in enzyme activity with depth also mirrored the changes in peat composition from the acrotelm to the catotelm. Season did play a role in temperature sensitivity with Ea of β-glucosidase and phosphatase being the lowest in August as expected but leucine amino peptidase (a nitrogen acquiring enzyme) Ea was not influenced by season. As temperatures rise, especially in winter months, enzymatic

  7. Analysis of Low-Biomass Microbial Communities in the Deep Biosphere.

    Science.gov (United States)

    Morono, Y; Inagaki, F

    2016-01-01

    Over the past few decades, the subseafloor biosphere has been explored by scientific ocean drilling to depths of about 2.5km below the seafloor. Although organic-rich anaerobic sedimentary habitats in the ocean margins harbor large numbers of microbial cells, microbial populations in ultraoligotrophic aerobic sedimentary habitats in the open ocean gyres are several orders of magnitude less abundant. Despite advances in cultivation-independent molecular ecological techniques, exploring the low-biomass environment remains technologically challenging, especially in the deep subseafloor biosphere. Reviewing the historical background of deep-biosphere analytical methods, the importance of obtaining clean samples and tracing contamination, as well as methods for detecting microbial life, technological aspects of molecular microbiology, and detecting subseafloor metabolic activity will be discussed. Copyright © 2016 Elsevier Inc. All rights reserved.

  8. Carbon use efficiency (CUE) and biomass turnover of soil microbial communities as affected by bedrock, land management and soil temperature and moisture

    Science.gov (United States)

    Zheng, Qing; Hu, Yuntao; Richter, Andreas; Wanek, Wolfgang

    2017-04-01

    Soil microbial carbon use efficiency (CUE), defined as the proportion of organic C taken up that is allocated to microbial growth, represents an important synthetic representation of microbial community C metabolism that describes the flux partitioning between microbial respiration and growth. Therefore, studying microbial CUE is critical for the understanding of soil C cycling. Microbial CUE is thought to vary with environmental conditions (e.g. temperature and soil moisture). Microbial CUE is thought to decrease with increasing temperature and declining soil moisture, as the latter may trigger stress responses (e.g. the synthesis of stress metabolites), which may consequently lower microbial community CUE. However, these effects on microbial CUE have not been adequately measured so far due to methodological restrictions. The most widely used methods for microbial CUE estimation are based on tracing 13C-labeled substrates into microbial biomass and respiratory CO2, approaches that are known to overestimate microbial CUE of native organic matter in soil. Recently, a novel substrate-independent approach based on the measurement of (i) respiration rates and (ii) the incorporation rates of 18O from labelled water into newly formed microbial DNA has been developed in our laboratory for measuring microbial CUE. This approach overcomes the shortcomings of previously used methods and has already been shown to yield realistic estimations of soil microbial CUE. This approach can also be applied to concurrently measure microbial biomass turnover rates, which also influence the sequestration of soil organic C. Microbial turnover rates are also thought to be impacted by environmental factors, but rarely have been directly measured so far. Here, we aimed at determining the short-term effects of environmental factors (soil temperature and soil moisture) on microbial CUE and microbial biomass turnover rates based on the novel 18O approach. Soils from three land-use types (arable

  9. Genome-centric metatranscriptomes and ecological roles of the active microbial populations during cellulosic biomass anaerobic digestion.

    Science.gov (United States)

    Jia, Yangyang; Ng, Siu-Kin; Lu, Hongyuan; Cai, Mingwei; Lee, Patrick K H

    2018-01-01

    Although anaerobic digestion for biogas production is used worldwide in treatment processes to recover energy from carbon-rich waste such as cellulosic biomass, the activities and interactions among the microbial populations that perform anaerobic digestion deserve further investigations, especially at the population genome level. To understand the cellulosic biomass-degrading potentials in two full-scale digesters, this study examined five methanogenic enrichment cultures derived from the digesters that anaerobically digested cellulose or xylan for more than 2 years under 35 or 55 °C conditions. Metagenomics and metatranscriptomics were used to capture the active microbial populations in each enrichment culture and reconstruct their meta-metabolic network and ecological roles. 107 population genomes were reconstructed from the five enrichment cultures using a differential coverage binning approach, of which only a subset was highly transcribed in the metatranscriptomes. Phylogenetic and functional convergence of communities by enrichment condition and phase of fermentation was observed for the highly transcribed populations in the metatranscriptomes. In the 35 °C cultures grown on cellulose, Clostridium cellulolyticum -related and Ruminococcus -related bacteria were identified as major hydrolyzers and primary fermenters in the early growth phase, while Clostridium leptum -related bacteria were major secondary fermenters and potential fatty acid scavengers in the late growth phase. While the meta-metabolism and trophic roles of the cultures were similar, the bacterial populations performing each function were distinct between the enrichment conditions. Overall, a population genome-centric view of the meta-metabolism and functional roles of key active players in anaerobic digestion of cellulosic biomass was obtained. This study represents a major step forward towards understanding the microbial functions and interactions at population genome level during the

  10. Continuous exposure of pesticides in an aquifer changes microbial biomass, diversity and degradation potential

    DEFF Research Database (Denmark)

    de Lipthay, J. R.; Johnsen, K.; Aamand, J.

    2000-01-01

    We studied in situ effects of pesticide exposure on microbial degradation potential and community structure of aquifer sediments. Sediment samples pre-exposed to pesticides were significantly different to non-exposed control samples. Pre-exposed sediment showed an increased degradation potential ...... towards phenoxyalcanoic acid herbicides as well as impact on microbial diversity was observed. Furthermore, bacterial biomass was changed, e.g. increased numbers of phenoxyalcanoic acid degraders in pesticide exposed sediment.......We studied in situ effects of pesticide exposure on microbial degradation potential and community structure of aquifer sediments. Sediment samples pre-exposed to pesticides were significantly different to non-exposed control samples. Pre-exposed sediment showed an increased degradation potential...

  11. Assimilable organic carbon (AOC in soil water extracts using Vibrio harveyi BB721 and its implication for microbial biomass.

    Directory of Open Access Journals (Sweden)

    Jincai Ma

    Full Text Available Assimilable organic carbon (AOC is commonly used to measure the growth potential of microorganisms in water, but has not yet been investigated for measuring microbial growth potential in soils. In this study, a simple, rapid, and non-growth based assay to determine AOC in soil was developed using a naturally occurring luminous strain Vibrio harveyi BB721 to determine the fraction of low molecular weight organic carbon in soil water extract. Calibration of the assay was achieved by measuring the luminescence intensity of starved V. harveyi BB721 cells in the late exponential phase with a concentration range from 0 to 800 µg l(-1 glucose (equivalent to 0-16.0 mg glucose C kg(-1 soil with the detection limit of 10 µg l(-1 equivalent to 0.20 mg glucose C kg(-1 soil. Results showed that bioluminescence was proportional to the concentration of glucose added to soil. The luminescence intensity of the cells was highly pH dependent and the optimal pH was about 7.0. The average AOC concentration in 32 soils tested was 2.9±2.2 mg glucose C kg(-1. Our data showed that AOC levels in soil water extracts were significantly correlated (P<0.05 with microbial biomass determined as microbial biomass carbon, indicating that the AOC concentrations determined by the method developed might be a good indicator of soil microbial biomass. Our findings provide a new approach that may be used to determine AOC in environmental samples using a non-growth bioluminescence based assay. Understanding the levels of AOC in soil water extract provides new insights into our ability to estimate the most available carbon pool to bacteria in soil that may be easily assimilated into cells for many metabolic processes and suggest possible the links between AOC, microbial regrowth potential, and microbial biomass in soils.

  12. Nitrogen fractions in the microbial biomass in soils of southern Brazil

    Directory of Open Access Journals (Sweden)

    F. A.O. Camargo

    1999-03-01

    Full Text Available The reaction of nitrogen compounds with ninhydrin can be used as an indicator of cytoplasmic materials released from microbial cells killed by fumigation. Total-N, ninhydrin-reactive-N (NR-N, ammonium-N (A-N, and α-amino-N in the microbial biomass of soils from the State of Rio Grande do Sul, Brazil, were determined, in 1996, in 0.5 mol L-1 K2SO4 extracts of fumigated and non-fumigated soils. Total-N varied from 20.3 to 104.4 mg kg-1 and the ninhydrin-reactive-N corresponded, in average, to 27% of this. The ninhydrin-reactive-N was made up of 67% ammonium-N and 33% aminoacids with the amino group at the α-carbon position. It was concluded that colorimetric analysis of NR-N and A-N may be used as a direct measure of microbial N in soil. This simple and rapid procedure is adequate for routine analyses.

  13. Colonization Habitat Controls Biomass, Composition, and Metabolic Activity of Attached Microbial Communities in the Columbia River Hyporheic Corridor

    Energy Technology Data Exchange (ETDEWEB)

    Stern, Noah; Ginder-Vogel, Matthew; Stegen, James C.; Arntzen, Evan; Kennedy, David W.; Larget, Bret R.; Roden, Eric E.; Kostka, Joel E.

    2017-06-09

    Hydrologic exchange plays a critical role in biogeochemical cycling within the hyporheic zone (the interface between river water and groundwater) of riverine ecosystems. Such exchange may set limits on the rates of microbial metabolism and impose deterministic selection on microbial communities that adapt to dynamically changing dissolved organic carbon (DOC) sources. This study examined the response of attached microbial communities (in situcolonized sand packs) from groundwater, hyporheic, and riverbed habitats within the Columbia River hyporheic corridor to “cross-feeding” with either groundwater, river water, or DOC-free artificial fluids. Our working hypothesis was that deterministic selection duringin situcolonization would dictate the response to cross-feeding, with communities displaying maximal biomass and respiration when supplied with their native fluid source. In contrast to expectations, the major observation was that the riverbed colonized sand had much higher biomass and respiratory activity, as well as a distinct community structure, compared with those of the hyporheic and groundwater colonized sands. 16S rRNA gene amplicon sequencing revealed a much higher proportion of certain heterotrophic taxa as well as significant numbers of eukaryotic algal chloroplasts in the riverbed colonized sand. Significant quantities of DOC were released from riverbed sediment and colonized sand, and separate experiments showed that the released DOC stimulated respiration in the groundwater and piezometer colonized sand. These results suggest that the accumulation and degradation of labile particulate organic carbon (POC) within the riverbed are likely to release DOC, which may enter the hyporheic corridor during hydrologic exchange, thereby stimulating microbial activity and imposing deterministic selective pressure on the microbial community composition.

    IMPORTANCEThe influence of river water

  14. Microbial nitrilases: versatile, spiral forming, industrial enzymes.

    Science.gov (United States)

    Thuku, R N; Brady, D; Benedik, M J; Sewell, B T

    2009-03-01

    The nitrilases are enzymes that convert nitriles to the corresponding acid and ammonia. They are members of a superfamily, which includes amidases and occur in both prokaryotes and eukaryotes. The superfamily is characterized by having a homodimeric building block with a alpha beta beta alpha-alpha beta beta alpha sandwich fold and an active site containing four positionally conserved residues: cys, glu, glu and lys. Their high chemical specificity and frequent enantioselectivity makes them attractive biocatalysts for the production of fine chemicals and pharmaceutical intermediates. Nitrilases are also used in the treatment of toxic industrial effluent and cyanide remediation. The superfamily enzymes have been visualized as dimers, tetramers, hexamers, octamers, tetradecamers, octadecamers and variable length helices, but all nitrilase oligomers have the same basic dimer interface. Moreover, in the case of the octamers, tetradecamers, octadecamers and the helices, common principles of subunit association apply. While the range of industrially interesting reactions catalysed by this enzyme class continues to increase, research efforts are still hampered by the lack of a high resolution microbial nitrilase structure which can provide insights into their specificity, enantioselectivity and the mechanism of catalysis. This review provides an overview of the current progress in elucidation of structure and function in this enzyme class and emphasizes insights that may lead to further biotechnological applications.

  15. Influence of diligent disintegration on anaerobic biomass and performance of microbial fuel cell.

    Science.gov (United States)

    Divyalakshmi, Palanisamy; Murugan, Devaraj; Rai, Chockalingam Lajapathi

    2017-12-01

    To enhance the performance of microbial fuel cells (MFC) by increasing the surface area of cathode and diligent mechanical disintegration of anaerobic biomass. Tannery effluent and anaerobic biomass were used. The increase in surface area of the cathode resulted in 78% COD removal, with the potential, current density, power density and coulombic efficiency of 675 mV, 147 mA m -2 , 33 mW m -2 and 3.5%, respectively. The work coupled with increased surface area of the cathode with diligent mechanical disintegration of the biomass, led to a further increase in COD removal of 82% with the potential, current density, power density and coulombic efficiency of 748 mV, 229 mA m -2 , 78 mW m -2 and 6% respectively. Mechanical disintegration of the biomass along with increased surface area of cathode enhances power generation in vertical MFC reactors using tannery effluent as fuel.

  16. Biomass assessment of microbial surface communities by means of hyperspectral remote sensing data.

    Science.gov (United States)

    Rodríguez-Caballero, Emilio; Paul, Max; Tamm, Alexandra; Caesar, Jennifer; Büdel, Burkhard; Escribano, Paula; Hill, Joachim; Weber, Bettina

    2017-05-15

    Dryland vegetation developed morphological and physiological strategies to cope with drought. However, as aridity increases, vascular plant coverage gets sparse and microbially-dominated surface communities (MSC), comprising cyanobacteria, algae, lichens and bryophytes together with heterotropic bacteria, archaea and fungi, gain relevance. Nevertheless, the relevance of MSC net primary productivity has only rarely been considered in ecosystem scale studies, and detailed information on their contribution to the total photosynthetic biomass reservoir is largely missing. In this study, we mapped the spatial distribution of two different MSC (biological soil crusts and quartz fields hosting hypolithic crusts) at two different sites within the South African Succulent Karoo (Soebatsfontein and Knersvlakte). Then we characterized both types of MSC in terms of chlorophyll content, and combining these data with the biocrust and quartz field maps, we estimated total biomass values of MSCs and their spatial patterns within the two different ecosystems. Our results revealed that MSC are important vegetation components of the South African Karoo biome, revealing clear differences between the two sites. At Soebatsfontein, MSC occurred as biological soil crusts (biocrusts), which covered about one third of the landscape reaching an overall biomass value of ~480gha -1 of chlorophyll a+b at the landscape scale. In the Knersvlakte, which is characterized by harsher environmental conditions (i.e. higher solar radiation and potential evapotranspiration), MSC occurred as biocrusts, but also formed hypolithic crusts growing on the lower soil-immersed parts of translucent quartz pebbles. Whereas chlorophyll concentrations of biocrusts and hypolithic crusts where insignificantly lower in the Knersvlakte, the overall MSC biomass reservoir was by far larger with ~780gha -1 of chlorophyll a+b. Thus, the complementary microbially-dominated surface communities promoted biomass formation within

  17. Taxonomic and Functional Responses of Soil Microbial Communities to Annual Removal of Aboveground Plant Biomass

    Science.gov (United States)

    Guo, Xue; Zhou, Xishu; Hale, Lauren; Yuan, Mengting; Feng, Jiajie; Ning, Daliang; Shi, Zhou; Qin, Yujia; Liu, Feifei; Wu, Liyou; He, Zhili; Van Nostrand, Joy D.; Liu, Xueduan; Luo, Yiqi; Tiedje, James M.; Zhou, Jizhong

    2018-01-01

    Clipping, removal of aboveground plant biomass, is an important issue in grassland ecology. However, few studies have focused on the effect of clipping on belowground microbial communities. Using integrated metagenomic technologies, we examined the taxonomic and functional responses of soil microbial communities to annual clipping (2010–2014) in a grassland ecosystem of the Great Plains of North America. Our results indicated that clipping significantly (P microbial respiration rates. Annual temporal variation within the microbial communities was much greater than the significant changes introduced by clipping, but cumulative effects of clipping were still observed in the long-term scale. The abundances of some bacterial and fungal lineages including Actinobacteria and Bacteroidetes were significantly (P microbial communities were significantly correlated with soil respiration and plant productivity. Intriguingly, clipping effects on microbial function may be highly regulated by precipitation at the interannual scale. Altogether, our results illustrated the potential of soil microbial communities for increased soil organic matter decomposition under clipping land-use practices. PMID:29904372

  18. Genome-scale biological models for industrial microbial systems.

    Science.gov (United States)

    Xu, Nan; Ye, Chao; Liu, Liming

    2018-04-01

    The primary aims and challenges associated with microbial fermentation include achieving faster cell growth, higher productivity, and more robust production processes. Genome-scale biological models, predicting the formation of an interaction among genetic materials, enzymes, and metabolites, constitute a systematic and comprehensive platform to analyze and optimize the microbial growth and production of biological products. Genome-scale biological models can help optimize microbial growth-associated traits by simulating biomass formation, predicting growth rates, and identifying the requirements for cell growth. With regard to microbial product biosynthesis, genome-scale biological models can be used to design product biosynthetic pathways, accelerate production efficiency, and reduce metabolic side effects, leading to improved production performance. The present review discusses the development of microbial genome-scale biological models since their emergence and emphasizes their pertinent application in improving industrial microbial fermentation of biological products.

  19. Bioaugmentation for Electricity Generation from Corn Stover Biomass Using Microbial Fuel Cells

    KAUST Repository

    Wang, Xin

    2009-08-01

    Corn stover is usually treated by an energy-intensive or expensive process to extract sugars for bioenergy production. However, it is possible to directly generate electricity from corn stover in microbial fuel cells (MFCs) through the addition of microbial consortia specifically acclimated for biomass breakdown. A mixed culture that was developed to have a high saccharification rate with corn stover was added to singlechamber, air-cathode MFCs acclimated for power production using glucose. The MFC produced a maximum power of 331 mW/ m 2 with the bioaugmented mixed culture and corn stover, compared to 510 mW/m2 using glucose. Denaturing gradient gel electrophoresis (DGGE) showed the communities continued to evolve on both the anode and corn stover biomass over 60 days, with several bacteria identified including Rhodopseudomonas palustris. The use of residual solids from the steam exploded corn stover produced 8% more power (406 mW/m2) than the raw corn stover. These results show that it is possible to directly generate electricity from waste corn stover in MFCs through bioaugmentation using naturally occurring bacteria. © 2009 American Chemical Society.

  20. Ultrasonic disintegration of microalgal biomass and consequent improvement of bioaccessibility/bioavailability in microbial fermentation.

    Science.gov (United States)

    Jeon, Byong-Hun; Choi, Jeong-A; Kim, Hyun-Chul; Hwang, Jae-Hoon; Abou-Shanab, Reda Ai; Dempsey, Brian A; Regan, John M; Kim, Jung Rae

    2013-01-01

    Microalgal biomass contains a high level of carbohydrates which can be biochemically converted to biofuels using state-of-the-art strategies that are almost always needed to employ a robust pretreatment on the biomass for enhanced energy production. In this study, we used an ultrasonic pretreatment to convert microalgal biomass (Scenedesmus obliquus YSW15) into feasible feedstock for microbial fermentation to produce ethanol and hydrogen. The effect of sonication condition was quantitatively evaluated with emphases on the characterization of carbohydrate components in microalgal suspension and on subsequent production of fermentative bioenergy. Scenedesmus obliquus YSW15 was isolated from the effluent of a municipal wastewater treatment plant. The sonication durations of 0, 10, 15, and 60 min were examined under different temperatures at a fixed frequency and acoustic power resulted in morphologically different states of microalgal biomass lysis. Fermentation was performed to evaluate the bioenergy production from the non-sonicated and sonicated algal biomasses after pretreatment stage under both mesophilic (35°C) and thermophilic (55°C) conditions. A 15 min sonication treatment significantly increased the concentration of dissolved carbohydrates (0.12 g g(-1)), which resulted in an increase of hydrogen/ethanol production through microbial fermentation. The bioconvertibility of microalgal biomass sonicated for 15 min or longer was comparable to starch as a control, indicating a high feasibility of using microalgae for fermentative bioenergy production. Increasing the sonication duration resulted in increases in both algal surface hydrophilicity and electrostatic repulsion among algal debris dispersed in aqueous solution. Scanning electron microscope images supported that ruptured algal cell allowed fermentative bacteria to access the inner space of the cell, evidencing an enhanced bioaccessibility. Sonication for 15 min was the best for fermentative

  1. Characterization of three plant biomass-degrading microbial consortia by metagenomics- and metasecretomics-based approaches

    NARCIS (Netherlands)

    Jiménez, Diego Javier; Brossi, Maria Julia de Lima; Schuckel, Julia; Kracun, Stjepan Kresimir; Willats, William George Tycho; van Elsas, Jan Dirk

    2016-01-01

    The selection of microbes by enrichment on plant biomass has been proposed as an efficient way to develop new strategies for lignocellulose saccharification. Here, we report an in-depth analysis of soil-derived microbial consortia that were trained to degrade once-used wheat straw (WS1-M),

  2. [Effects of adding straw carbon source to root knot nematode diseased soil on soil microbial biomass and protozoa abundance].

    Science.gov (United States)

    Zhang, Si-Hui; Lian, Jian-Hong; Cao, Zhi-Ping; Zhao, Li

    2013-06-01

    A field experiment with successive planting of tomato was conducted to study the effects of adding different amounts of winter wheat straw (2.08 g x kg(-1), 1N; 4.16 g x kg(-1), 2N; and 8.32 g x kg(-1), 4N) to the soil seriously suffered from root knot nematode disease on the soil microbial biomass and protozoa abundance. Adding straw carbon source had significant effects on the contents of soil microbial biomass carbon (MBC) and microbial biomass nitrogen (MBN) and the abundance of soil protozoa, which all decreased in the order of 4N > 2N > 1N > CK. The community structure of soil protozoa also changed significantly under straw addition. In the treatments with straw addition, the average proportion of fagellate, amoeba, and ciliates accounted for 36.0%, 59.5%, and 4.5% of the total protozoa, respectively. Under the same adding amounts of wheat straw, there was an increase in the soil MBC and MBN contents, MBC/MBN ratio, and protozoa abundance with increasing cultivation period.

  3. Soil microbial species loss affects plant biomass and survival of an introduced bacterial strain, but not inducible plant defences.

    Science.gov (United States)

    Kurm, Viola; van der Putten, Wim H; Pineda, Ana; Hol, W H Gera

    2018-02-12

    Plant growth-promoting rhizobacteria (PGPR) strains can influence plant-insect interactions. However, little is known about the effect of changes in the soil bacterial community in general and especially the loss of rare soil microbes on these interactions. Here, the influence of rare soil microbe reduction on induced systemic resistance (ISR) in a wild ecotype of Arabidopsis thaliana against the aphid Myzus persicae was investigated. To create a gradient of microbial abundances, soil was inoculated with a serial dilution of a microbial community and responses of Arabidopsis plants that originated from the same site as the soil microbes were tested. Plant biomass, transcription of genes involved in plant defences, and insect performance were measured. In addition, the effects of the PGPR strain Pseudomonas fluorescens SS101 on plant and insect performance were tested under the influence of the various soil dilution treatments. Plant biomass showed a hump-shaped relationship with soil microbial community dilution, independent of aphid or Pseudomonas treatments. Both aphid infestation and inoculation with Pseudomonas reduced plant biomass, and led to downregulation of PR1 (salicylic acid-responsive gene) and CYP79B3 (involved in synthesis of glucosinolates). Aphid performance and gene transcription were unaffected by soil dilution. Neither the loss of rare microbial species, as caused by soil dilution, nor Pseudomonas affect the resistance of A. thaliana against M. persicae. However, both Pseudomonas survival and plant biomass respond to rare species loss. Thus, loss of rare soil microbial species can have a significant impact on both above- and below-ground organisms. © The Author(s) 2018. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  4. Short- and long-term effects of nutrient enrichment on microbial exoenzyme activity in mangrove peat

    KAUST Repository

    Keuskamp, Joost A.; Feller, Ilka C.; Laanbroek, Hendrikus J.; Verhoeven, Jos T.A.; Hefting, Mariet M.

    2015-01-01

    -limited mangroves. To examine this, we quantified the short- and long-term effects of N and P enrichment on microbial biomass and decomposition-related enzyme activities in a Rhizophora mangle-dominated mangrove, which had been subjected to fertilisation treatments

  5. Enzyme activities by indicator of quality in organic soil

    Science.gov (United States)

    Raigon Jiménez, Mo; Fita, Ana Delores; Rodriguez Burruezo, Adrián

    2016-04-01

    The analytical determination of biochemical parameters, as soil enzyme activities and those related to the microbial biomass is growing importance by biological indicator in soil science studies. The metabolic activity in soil is responsible of important processes such as mineralization and humification of organic matter. These biological reactions will affect other key processes involved with elements like carbon, nitrogen and phosphorus , and all transformations related in soil microbial biomass. The determination of biochemical parameters is useful in studies carried out on organic soil where microbial processes that are key to their conservation can be analyzed through parameters of the metabolic activity of these soils. The main objective of this work is to apply analytical methodologies of enzyme activities in soil collections of different physicochemical characteristics. There have been selective sampling of natural soils, organic farming soils, conventional farming soils and urban soils. The soils have been properly identified conserved at 4 ° C until analysis. The enzyme activities determinations have been: catalase, urease, cellulase, dehydrogenase and alkaline phosphatase, which bring together a representative group of biological transformations that occur in the soil environment. The results indicate that for natural and agronomic soil collections, the values of the enzymatic activities are within the ranges established for forestry and agricultural soils. Organic soils are generally higher level of enzymatic, regardless activity of the enzyme involved. Soil near an urban area, levels of activities have been significantly reduced. The vegetation cover applied to organic soils, results in greater enzymatic activity. So the quality of these soils, defined as the ability to maintain their biological productivity is increased with the use of cover crops, whether or spontaneous species. The practice of cover based on legumes could be used as an ideal choice

  6. Hydrolytic enzyme activity enhanced by Barium supplementation

    Directory of Open Access Journals (Sweden)

    Camilo Muñoz

    2016-10-01

    Full Text Available Hydrolysis of polymers is a first and often limiting step during the degradation of plant residues. Plant biomass is generally a major component of waste residues and a major renewable resource to obtain a variety of secondary products including biofuels. Improving the performance of enzymatic hydrolysis of plant material with minimum costs and limiting the use of additional microbial biomass or hydrolytic enzymes directly influences competitiveness of these green biotechnological processes. In this study, we cloned and expressed a cellulase and two esterases recovered from environmental thermophilic soil bacterial communities and characterize their optimum activity conditions including the effect of several metal ions. Results showed that supplementing these hydrolytic reactions with Barium increases the activity of these extracellular hydrolytic enzymes. This observation represents a simple but major improvement to enhance the efficiency and competitiveness of this process within an increasingly important biotechnological sector.

  7. Evaluation of gastrointestinal bacterial population for the production of holocellulose enzymes for biomass deconstruction.

    Science.gov (United States)

    Asem, Dhaneshwaree; Leo, Vincent Vineeth; Passari, Ajit Kumar; Tonsing, Mary Vanlalhruaii; Joshi, J Beslin; Uthandi, Sivakumar; Hashem, Abeer; Abd Allah, Elsayed Fathi; Singh, Bhim Pratap

    2017-01-01

    The gastrointestinal (GI) habitat of ruminant and non-ruminant animals sustains a vast ensemble of microbes that are capable of utilizing lignocellulosic plant biomass. In this study, an indigenous swine (Zovawk) and a domesticated goat (Black Bengal) were investigated to isolate bacteria having plant biomass degrading enzymes. After screening and enzymatic quantification of eighty-one obtained bacterial isolates, Serratia rubidaea strain DBT4 and Aneurinibacillus aneurinilyticus strain DBT87 were revealed as the most potent strains, showing both cellulase and xylanase production. A biomass utilization study showed that submerged fermentation (SmF) of D2 (alkaline pretreated pulpy biomass) using strain DBT4 resulted in the most efficient biomass deconstruction with maximum xylanase (11.98 U/mL) and FPase (0.5 U/mL) activities (55°C, pH 8). The present study demonstrated that bacterial strains residing in the gastrointestinal region of non-ruminant swine are a promising source for lignocellulose degrading microorganisms that could be used for biomass conversion.

  8. Evaluation of gastrointestinal bacterial population for the production of holocellulose enzymes for biomass deconstruction.

    Directory of Open Access Journals (Sweden)

    Dhaneshwaree Asem

    Full Text Available The gastrointestinal (GI habitat of ruminant and non-ruminant animals sustains a vast ensemble of microbes that are capable of utilizing lignocellulosic plant biomass. In this study, an indigenous swine (Zovawk and a domesticated goat (Black Bengal were investigated to isolate bacteria having plant biomass degrading enzymes. After screening and enzymatic quantification of eighty-one obtained bacterial isolates, Serratia rubidaea strain DBT4 and Aneurinibacillus aneurinilyticus strain DBT87 were revealed as the most potent strains, showing both cellulase and xylanase production. A biomass utilization study showed that submerged fermentation (SmF of D2 (alkaline pretreated pulpy biomass using strain DBT4 resulted in the most efficient biomass deconstruction with maximum xylanase (11.98 U/mL and FPase (0.5 U/mL activities (55°C, pH 8. The present study demonstrated that bacterial strains residing in the gastrointestinal region of non-ruminant swine are a promising source for lignocellulose degrading microorganisms that could be used for biomass conversion.

  9. Assessment of tillage systems in organic farming: influence of soil structure on microbial biomass. First results

    OpenAIRE

    Vian, Jean François; Peigné, Joséphine; Chaussod, Rémi; Roger-Estrade, Jean

    2007-01-01

    Soil tillage modifies environmental conditions of soil microorganisms and their ability to release nitrogen. We compare the influence of reduced tillage (RT) and mouldboard ploughing (MP) on the soil microbial functioning in organic farming. In order to connect soil structure generated by these tillage systems on the soil microbial biomass we adopt a particular sampling scheme based on the morphological characterisation of the soil structure by the description of the soil profile. This method...

  10. Imidacloprid application changes microbial dynamics and enzymes in rice soil.

    Science.gov (United States)

    Mahapatra, Bibhab; Adak, Totan; Patil, Naveen K B; Pandi G, Guru P; Gowda, G Basana; Jambhulkar, N N; Yadav, Manoj Kumar; Panneerselvam, P; Kumar, Upendra; Munda, Sushmita; Jena, Mayabini

    2017-10-01

    Extensive use of imidacloprid in rice ecosystem may alter dynamics of microorganisms and can change soil biochemical properties. The objective of this study was to assess the effect of imidacloprid on growth and activities of microbes in tropical rice soil ecosystem. Four treatments, namely, recommended dose (at 25g a.i. ha -1 , RD), double the recommended dose (at 50g a.i. ha -1 , 2RD), five times the recommended dose (at 125g a.i. ha -1 , 5RD) & ten times the recommended dose (at 250g a.i. ha -1 , 10RD) along with control were imposed under controlled condition. Dissipation half lives of imidacloprid in soil were 19.25, 20.38, 21.65 and 33.00 days for RD, 2RD, 5RD and 10RD, respectively. In general bacteria, actinomycetes, fungi and phosphate solubilising bacteria population were disturbed due to imidacloprid application. Changes in diversity indices within bacterial community confirmed that imidacloprid application significantly affected distribution of bacteria. Total soil microbial biomass carbon content was reduced on imidacloprid application. Except dehydrogenase and alkaline phosphatase activities, all other soil enzymes namely, β-glycosidase, fluorescien diacetate hydrolase, acid phosphatase and urease responded negatively to imidacloprid application. The extent of negative effect of imidacloprid depends on dose and exposure time. This study concludes imidacloprid application had transient negative effects on soil microbes. Copyright © 2017 Elsevier Inc. All rights reserved.

  11. High levels of maize in broiler diets with or without microbial enzyme ...

    African Journals Online (AJOL)

    mbhuiya3

    2013-03-13

    Mar 13, 2013 ... High levels of maize in broiler diets with or without microbial enzyme .... to improve carbohydrate digestion and availability of phosphorus from ... grinding machine and stored at –4 ºC in airtight containers for chemical analysis ...

  12. Bacterial synergism in lignocellulose biomass degradation : Complementary roles of degraders as influenced by complexity of the carbon source

    NARCIS (Netherlands)

    Cortes Tolalpa, Larisa; Falcao Salles, Joana; van Elsas, Jan

    2017-01-01

    Lignocellulosic biomass (LCB) is an attractive source of carbon for the production of sugars and other chemicals. Due to its inherent complexity and heterogeneity, efficient biodegradation requires the actions of different types of hydrolytic enzymes. In nature, complex microbial communities that

  13. Spatial distribution of enzyme driven reactions at micro-scales

    Science.gov (United States)

    Kandeler, Ellen; Boeddinghaus, Runa; Nassal, Dinah; Preusser, Sebastian; Marhan, Sven; Poll, Christian

    2017-04-01

    Studies of microbial biogeography can often provide key insights into the physiologies, environmental tolerances, and ecological strategies of soil microorganisms that dominate in natural environments. In comparison with aquatic systems, soils are particularly heterogeneous. Soil heterogeneity results from the interaction of a hierarchical series of interrelated variables that fluctuate at many different spatial and temporal scales. Whereas spatial dependence of chemical and physical soil properties is well known at scales ranging from decimetres to several hundred metres, the spatial structure of soil enzymes is less clear. Previous work has primarily focused on spatial heterogeneity at a single analytical scale using the distribution of individual cells, specific types of organisms or collective parameters such as bacterial abundance or total microbial biomass. There are fewer studies that have considered variations in community function and soil enzyme activities. This presentation will give an overview about recent studies focusing on spatial pattern of different soil enzymes in the terrestrial environment. Whereas zymography allows the visualization of enzyme pattern in the close vicinity of roots, micro-sampling strategies followed by MUF analyses clarify micro-scale pattern of enzymes associated to specific microhabitats (micro-aggregates, organo-mineral complexes, subsoil compartments).

  14. Linking temperature sensitivity of soil organic matter decomposition to its molecular structure, accessibility, and microbial physiology.

    Science.gov (United States)

    Wagai, Rota; Kishimoto-Mo, Ayaka W; Yonemura, Seiichiro; Shirato, Yasuhito; Hiradate, Syuntaro; Yagasaki, Yasumi

    2013-04-01

    Temperature sensitivity of soil organic matter (SOM) decomposition may have a significant impact on global warming. Enzyme-kinetic hypothesis suggests that decomposition of low-quality substrate (recalcitrant molecular structure) requires higher activation energy and thus has greater temperature sensitivity than that of high-quality, labile substrate. Supporting evidence, however, relies largely on indirect indices of substrate quality. Furthermore, the enzyme-substrate reactions that drive decomposition may be regulated by microbial physiology and/or constrained by protective effects of soil mineral matrix. We thus tested the kinetic hypothesis by directly assessing the carbon molecular structure of low-density fraction (LF) which represents readily accessible, mineral-free SOM pool. Using five mineral soil samples of contrasting SOM concentrations, we conducted 30-days incubations (15, 25, and 35 °C) to measure microbial respiration and quantified easily soluble C as well as microbial biomass C pools before and after the incubations. Carbon structure of LFs (soil was measured by solid-state (13) C-NMR. Decomposition Q10 was significantly correlated with the abundance of aromatic plus alkyl-C relative to O-alkyl-C groups in LFs but not in bulk soil fraction or with the indirect C quality indices based on microbial respiration or biomass. The warming did not significantly change the concentration of biomass C or the three types of soluble C despite two- to three-fold increase in respiration. Thus, enhanced microbial maintenance respiration (reduced C-use efficiency) especially in the soils rich in recalcitrant LF might lead to the apparent equilibrium between SOM solubilization and microbial C uptake. Our results showed physical fractionation coupled with direct assessment of molecular structure as an effective approach and supported the enzyme-kinetic interpretation of widely observed C quality-temperature relationship for short-term decomposition. Factors

  15. Mechanisms of microbial destabilization of soil C shifts over decades of warming

    Science.gov (United States)

    DeAngelis, K.; Pold, G.; Chowdhury, P. R.; Schnabel, J.; Grandy, S.; Melillo, J. M.

    2017-12-01

    Microbes are major actors in regulating the earth's biogeochemical cycles, with temperature-sensitive microbial tradeoffs improving ecosystem biogeochemical models. Meanwhile, the Earth's climate is changing, with decades of warming undercutting the ability of soil to store carbon. Our work explores trends of 26 years of experimental warming in temperate deciduous forest soils, which is associated with cycles of soil carbon degradation punctuated by periods of changes in soil microbial dynamics. Using a combination of biogeochemistry and molecular analytical methods, we explore the hypotheses that substrate availability, community structure, altered temperature sensitivity of microbial turnover-growth efficiency tradeoff, and microbial evolution are responsible for observations of accelerated degradation of soil carbon over time. Amplicon sequencing of microbial communities suggests a small role of changing microbial community composition over decades of warming, but a sustained suppression of fungal biomass is accompanied by increased biomass of Actinobacteria, Actinobacteria, Alphaproteobacteria, Verrucomicrobia and Planctomycetes. Substrate availability plays an important role in microbial dynamics, with depleted labile carbon in the first decade and depleted lignin in the second decade. Increased lignin-degrading enzyme activity supports the suggestion that lignin-like organic matter is an important substrate in chronically warmed soils. Metatranscriptomics data support the suggestion that increased turnover is associated with long-term warming, with metagenomic signals of increased carbohydrate-degrading enzymes in the organic horizon but decreased in the mineral soils. Finally, traits analysis of over 200 cultivated isolates of bacterial species from heated and control soils suggests an expanded ability for degradation of cellulose and hemicellulose but not chitin, supporting the hypothesis that long-term warming is exerting evolutionary pressure on microbial

  16. Microbial P450 Enzymes in Bioremediation and Drug Discovery: Emerging Potentials and Challenges.

    Science.gov (United States)

    Bhattacharya, Sukanta S; Yadav, Jagjit S

    2018-01-01

    Cytochrome P450 enzymes are a structurally conserved but functionally diverse group of heme-containing mixed function oxidases found across both prokaryotic and eukaryotic forms of the microbial world. Microbial P450s are known to perform diverse functions ranging from the synthesis of cell wall components to xenobiotic/drug metabolism to biodegradation of environmental chemicals. Conventionally, many microbial systems have been reported to mimic mammalian P450-like activation of drugs and were proposed as the in-vitro models of mammalian drug metabolism. Recent reports suggest that native or engineered forms of specific microbial P450s from these and other microbial systems could be employed for desired specific biotransformation reactions toward natural and synthetic (drug) compounds underscoring their emerging potential in drug improvement and discovery. On the other hand, microorganisms particularly fungi and actinomycetes have been shown to possess catabolic P450s with unusual potential to degrade toxic environmental chemicals including persistent organic pollutants (POPs). Wood-rotting basidiomycete fungi in particular have revealed the presence of exceptionally large P450 repertoire (P450ome) in their genomes, majority of which are however orphan (with no known function). Our pre- and post-genomic studies have led to functional characterization of several fungal P450s inducible in response to exposure to several environmental toxicants and demonstration of their potential in bioremediation of these chemicals. This review is an attempt to summarize the postgenomic unveiling of this versatile enzyme superfamily in microbial systems and investigation of their potential to synthesize new drugs and degrade persistent pollutants, among other biotechnological applications. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  17. Development of over-production strain of saccharification enzyme and biomass pretreatment by proton beam irradiation

    International Nuclear Information System (INIS)

    Kim, S. W.; Lee, J. Y.; Song, Y. S.; Lee, S. J.; Shin, H. Y.; Kim, S. B.

    2010-04-01

    When lignocellulosic biomass converts to ethanol, enzyme takes lots of part of whole cost. Therefore, cellulase production is one of the important processes for the successful enzymatic conversion of cellulosic biomass to ethanol. Among cellulolytic enzymes, cellulase is multi-complex enzyme containing endo-glucanase, exo-glucanase and β-glucosidase. Cellulolyticfungi, Trichodema reesei is well known to produce the highest yields of cellulase. Especially, suitable cellulase composition was important for the effective saccharification of lignocellulosic biomass and strain having high level production of cellulase should be developed for hydrolysis. For efficient ethanol production, hemicellullase of Aspergillus also develop to use xylose generated from saccharification of biomass. In this study, pretreatment process of rice straw using proton beam irradiation (PBI) was carried out for enhancement of enzyme digestibility at different proton beam doses. Also, PBI pretreatment on ammonia soaking treated (SAA, Soaking aqueous ammonia) rice straw was conducted to solve the problem that is micro-structural inhibition of rice straw. Optimal dosages of proton beam on rice straw and SAA treated rice straw for efficient recovery of sugar were 15 KGy and 3 KGy, respectively. Enzymatic saccharification of PBI treated rice straw and SAA rice straw was conducted for the guidance of NREL standard procedure. Analysis using X-ray diffractometry (XRD) for crystallinity index was carried out and CrI found to be 33.38% of control and 35.72% of 15 KGy. Also, CrI was determined to be 67.11% of control and approximately 65.58% of 3 kGy dose in PBI pretreatment on SAA treated rice straw. The result of sugar recovery of both was approximately 70 % and 91 % of theoretical glucose contents, respectively. The initial reaction rate was increased from 7.610 -4 g·l -1 ·s -1 of 15 KGy (PBI pretreated rice straw) to 9.710 -4 g·l -1 ·s -1 (3 KGy PBI pretreated SAA rice straw). The selection of

  18. On polydispersity of plant biomass recalcitrance and its effects on pretreatment optimization for sugar production

    Science.gov (United States)

    J.Y. Zhu; Steve P. Verrill; Hao Liu; Victoria L. Herian; Xuejun Pan; Donald L. Rockwood

    2011-01-01

    This paper discusses a property associated with plant biomass recalcitrance to enzyme and microbial deconstructions in sugar production from cellulose and hemicelluloses. The hemicelluloses are more readily hydrolyzed to sugars than is cellulose. As a result, optimization to maximize individual glucose and hemicellulose sugar recovery is not possible. This property is...

  19. Abrolhos bank reef health evaluated by means of water quality, microbial diversity, benthic cover, and fish biomass data.

    Directory of Open Access Journals (Sweden)

    Thiago Bruce

    Full Text Available The health of the coral reefs of the Abrolhos Bank (Southwestern Atlantic was characterized with a holistic approach using measurements of four ecosystem components: (i inorganic and organic nutrient concentrations, [1] fish biomass, [1] macroalgal and coral cover and (iv microbial community composition and abundance. The possible benefits of protection from fishing were particularly evaluated by comparing sites with varying levels of protection. Two reefs within the well-enforced no-take area of the National Marine Park of Abrolhos (Parcel dos Abrolhos and California were compared with two unprotected coastal reefs (Sebastião Gomes and Pedra de Leste and one legally protected but poorly enforced coastal reef (the "paper park" of Timbebas Reef. The fish biomass was lower and the fleshy macroalgal cover was higher in the unprotected reefs compared with the protected areas. The unprotected and protected reefs had similar seawater chemistry. Lower vibrio CFU counts were observed in the fully protected area of California Reef. Metagenome analysis showed that the unprotected reefs had a higher abundance of archaeal and viral sequences and more bacterial pathogens, while the protected reefs had a higher abundance of genes related to photosynthesis. Similar to other reef systems in the world, there was evidence that reductions in the biomass of herbivorous fishes and the consequent increase in macroalgal cover in the Abrolhos Bank may be affecting microbial diversity and abundance. Through the integration of different types of ecological data, the present study showed that protection from fishing may lead to greater reef health. The data presented herein suggest that protected coral reefs have higher microbial diversity, with the most degraded reef (Sebastião Gomes showing a marked reduction in microbial species richness. It is concluded that ecological conditions in unprotected reefs may promote the growth and rapid evolution of opportunistic

  20. Alkaline peroxide pretreatment of corn stover: effects of biomass, peroxide, and enzyme loading and composition on yields of glucose and xylose

    Science.gov (United States)

    2011-01-01

    Background Pretreatment is a critical step in the conversion of lignocellulose to fermentable sugars. Although many pretreatment processes are currently under investigation, none of them are entirely satisfactory in regard to effectiveness, cost, or environmental impact. The use of hydrogen peroxide at pH 11.5 (alkaline hydrogen peroxide (AHP)) was shown by Gould and coworkers to be an effective pretreatment of grass stovers and other plant materials in the context of animal nutrition and ethanol production. Our earlier experiments indicated that AHP performed well when compared against two other alkaline pretreatments. Here, we explored several key parameters to test the potential of AHP for further improvement relevant to lignocellulosic ethanol production. Results The effects of biomass loading, hydrogen peroxide loading, residence time, and pH control were tested in combination with subsequent digestion with a commercial enzyme preparation, optimized mixtures of four commercial enzymes, or optimized synthetic mixtures of pure enzymes. AHP pretreatment was performed at room temperature (23°C) and atmospheric pressure, and after AHP pretreatment the biomass was neutralized with HCl but not washed before enzyme digestion. Standard enzyme digestion conditions were 0.2% glucan loading, 15 mg protein/g glucan, and 48 h digestion at 50°C. Higher pretreatment biomass loadings (10% to 20%) gave higher monomeric glucose (Glc) and xylose (Xyl) yields than the 2% loading used in earlier studies. An H2O2 loading of 0.25 g/g biomass was almost as effective as 0.5 g/g, but 0.125 g/g was significantly less effective. Optimized mixtures of four commercial enzymes substantially increased post-AHP-pretreatment enzymatic hydrolysis yields at all H2O2 concentrations compared to any single commercial enzyme. At a pretreatment biomass loading of 10% and an H2O2 loading of 0.5 g/g biomass, an optimized commercial mixture at total protein loadings of 8 or 15 mg/g glucan gave

  1. Photosynthetic microbial desalination cells (PMDCs) for clean energy, water and biomass production.

    Science.gov (United States)

    Kokabian, Bahareh; Gude, Veera Gnaneswar

    2013-12-01

    Current microbial desalination cell (MDC) performances are evaluated with chemical catalysts such as ferricyanide, platinum catalyzed air-cathodes or aerated cathodes. All of these methods improve power generation potential in MDCs, however, they are not preferable for large scale applications due to cost, energy and environmental toxicity issues. In this study, performance of microbial desalination cells with an air cathode and an algae biocathode (Photosynthetic MDC - PMDC) were evaluated, both under passive conditions (no mechanical aeration or mixing). The results indicate that passive algae biocathodes perform better than air cathodes and enhance COD removal and utilize treated wastewater as the growth medium to obtain valuable biomass for high value bioproducts. Maximum power densities of 84 mW m(-3) (anode volume) or 151 mW m(-3) (biocathode volume) and a desalination rate of 40% were measured with 0.9 : 1 : 0.5 volumetric ratios of anode, desalination and algae biocathode chambers respectively. This first proof-of-concept study proves that the passive mechanisms can be beneficial in enhancing the sustainability of microbial desalination cells.

  2. Microbial biomass and biological activity of soils and soil-like bodies in coastal oases of Antarctica

    Science.gov (United States)

    Nikitin, D. A.; Marfenina, O. E.; Kudinova, A. G.; Lysak, L. V.; Mergelov, N. S.; Dolgikh, A. V.; Lupachev, A. V.

    2017-09-01

    The method of luminescent microscopy has been applied to study the structure of the microbial biomass of soils and soil-like bodies in East (the Thala Hills and Larsemann Hills oases) and West (Cape Burks, Hobbs coast) Antarctica. According to Soil Taxonomy, the studied soils mainly belong to the subgroups of Aquic Haploturbels, Typic Haploturbels, Typic Haplorthels, and Lithic Haplorthels. The major contribution to their microbial biomass belongs to fungi. The highest fungal biomass (up to 790 μg C/g soil) has been found in the soils with surface organic horizons in the form of thin moss/lichen litters, in which the development of fungal mycelium is most active. A larger part of fungal biomass (70-98%) is represented by spores. For the soils without vegetation cover, the accumulation of bacterial and fungal biomass takes place in the horizons under surface desert pavements. In the upper parts of the soils without vegetation cover and in the organic soil horizons, the major part (>60%) of fungal mycelium contains protective melanin pigments. Among bacteria, the high portion (up to 50%) of small filtering forms is observed. A considerable increase (up to 290.2 ± 27 μg C/g soil) in the fungal biomass owing to the development of yeasts has been shown for gley soils (gleyzems) developing from sapropel sediments under subaquatic conditions and for the algal-bacterial mat on the bottom of the lake (920.7 ± 46 μg C/g soil). The production of carbon dioxide by the soils varies from 0.47 to 2.34 μg C-CO2/(g day). The intensity of nitrogen fixation in the studied samples is generally low: from 0.08 to 55.85 ng C2H4/(g day). The intensity of denitrification varies from 0.09 to 19.28 μg N-N2O/(g day).

  3. Mineralogical controls on microbial biomass accumulation on two tropical soils

    Science.gov (United States)

    Block, K. A.; Pena, S. A.; Katz, A.; Gottlieb, P.; Volta, A.

    2017-12-01

    The characteristics of soil organic matter (SOM) generated by microbes and associated with minerals are not well defined. This information is critical to reducing uncertainty in climate models related to C cycling and ecosystem feedbacks. The resistance to degradation of mineral-associated SOM is influenced by aggregate structure, mineral chemistry and microbial community. In this work we examine the influence of mineral composition, including amorphous coatings on the biomass yield and aggregate structure through thermogravimetric analysis, X-ray diffraction and electron microscopy. Two soil organisms, Pseudomonas phaseolicola, and Streptomyces griseosporus, were each incubated over a 72-hour period in minimal media with the cultured under the same conditions. In all samples, approximately half of the sample mass loss occurred between 175 ºC - 375 ºC, which we attribute to biomolecules accumulated on the mineral surfaces. We observed a slightly larger mass loss in the Inceptisol than in the Oxisol, most of which corresponded to compounds that underwent pyrolysis at 300 ºC. HRTEM micrographs and TEM-EDS image maps showing the spatial relationship of microbial necromass to soil minerals will be reported.

  4. Effects of lead and cadmium nitrate on biomass and substrate utilization pattern of soil microbial communities.

    Science.gov (United States)

    Muhammad, Akmal; Xu, Jianming; Li, Zhaojun; Wang, Haizhen; Yao, Huaiying

    2005-07-01

    A study was conducted to evaluate the effects of different concentrations of lead (Pb) and cadmium (Cd) applied as their nitrates on soil microbial biomass carbon (C(mic)) and nitrogen (N(mic)), and substrate utilization pattern of soil microbial communities. The C(mic) and N(mic) contents were determined at 0, 14, 28, 42 and 56 days after heavy metal application (DAA). The results showed a significant decline in the C(mic) for all Pb and Cd amended soils from the start to 28 DAA. From 28 to 56 DAA, C(mic) contents changed non-significantly for all other treatments except for 600 mgkg(-1) Pb and 100 mgkg(-1) Cd in which it declined significantly from 42 to 56 DAA. The N(mic) contents also decreased significantly from start to 28 DAA for all other Pb and Cd treatments except for 200 mgkg(-1) Pb which did not show significant difference from the control. Control and 200 mgkg(-1) Pb had significantly lower soil microbial biomass C:N ratio as compared with other Pb treatments from 14 to 42 DAA, however at 56 DAA, only 1000 mgkg(-1) Pb showed significantly higher C:N ratio compared with other treatments. No significant difference in C:N ratio for all Cd treated soils was seen from start to 28 DAA, however from 42 to 56 DAA, 100 mgkg(-1) Pb showed significantly higher C:N ratio compared with other treatments. On 56 DAA, substrate utilization pattern of soil microbial communities was determined by inoculating Biolog ECO plates. The results indicated that Pb and Cd addition inhibited the functional activity of soil microbial communities as indicated by the intensity of average well color development (AWCD) during 168 h of incubation. Multivariate analysis of sole carbon source utilization pattern demonstrated that higher levels of heavy metal application had significantly affected soil microbial community structure.

  5. Earthworms (Amynthas spp. increase common bean growth, microbial biomass, and soil respiration

    Directory of Open Access Journals (Sweden)

    Julierme Zimmer Barbosa

    2017-10-01

    Full Text Available Few studies have evaluated the effect of earthworms on plants and biological soil attributes, especially among legumes. The objective of this study was to evaluate the influence of earthworms (Amynthas spp. on growth in the common bean (Phaseolus vulgaris L. and on soil biological attributes. The experiment was conducted in a greenhouse using a completely randomized design with five treatments and eight repetitions. The treatments consisted of inoculation with five different quantities of earthworms of the genus Amynthas (0, 2, 4, 6, and 8 worms per pot. Each experimental unit consisted of a plastic pot containing 4 kg of soil and two common bean plants. The experiment was harvested 38 days after seedling emergence. Dry matter and plant height, soil respiration, microbial respiration, microbial biomass, and metabolic quotient were determined. Earthworm recovery in our study was high in number and mass, with all values above 91.6% and 89.1%, respectively. In addition, earthworm fresh biomass decreased only in the treatment that included eight earthworms per pot. The presence of earthworms increased the plant growth and improved soil biological properties, suggesting that agricultural practices that favor the presence of these organisms can be used to increase the production of common bean, and the increased soil CO2 emission caused by the earthworms can be partially offset by the addition of common bean crop residues to the soil.

  6. Metatranscriptomics Reveals the Functions and Enzyme Profiles of the Microbial Community in Chinese Nong-Flavor Liquor Starter

    Directory of Open Access Journals (Sweden)

    Yuhong Huang

    2017-09-01

    Full Text Available Chinese liquor is one of the world's best-known distilled spirits and is the largest spirit category by sales. The unique and traditional solid-state fermentation technology used to produce Chinese liquor has been in continuous use for several thousand years. The diverse and dynamic microbial community in a liquor starter is the main contributor to liquor brewing. However, little is known about the ecological distribution and functional importance of these community members. In this study, metatranscriptomics was used to comprehensively explore the active microbial community members and key transcripts with significant functions in the liquor starter production process. Fungi were found to be the most abundant and active community members. A total of 932 carbohydrate-active enzymes, including highly expressed auxiliary activity family 9 and 10 proteins, were identified at 62°C under aerobic conditions. Some potential thermostable enzymes were identified at 50, 62, and 25°C (mature stage. Increased content and overexpressed key enzymes involved in glycolysis and starch, pyruvate and ethanol metabolism were detected at 50 and 62°C. The key enzymes of the citrate cycle were up-regulated at 62°C, and their abundant derivatives are crucial for flavor generation. Here, the metabolism and functional enzymes of the active microbial communities in NF liquor starter were studied, which could pave the way to initiate improvements in liquor quality and to discover microbes that produce novel enzymes or high-value added products.

  7. Influence of soil management practices and substrate availability on microbial biomass and its activities in some haplic luvisols

    Energy Technology Data Exchange (ETDEWEB)

    Friedel, Jurgen K [University Hohenheeim, Stuttgart (Germany)

    1996-07-01

    Soil microbial biomass and activities are sensitive indicators of management effects. Higher contents of microbial biomass and higher activities, for example, are found with crop rotations in contrast to bare fallow and mono culture systems. The main reason for these differences is a higher input of crop and root residues in crop rotation systems, leading to more microbial available substrate. The objectives of this study were to describe indices for microbial available substrate in arable soils depending on management practices, and to relate them with soil microbial biomass and activities. At two locations (Muttergarten and hinger Hof near the University of Hohenheim, Stuttgart, SW-Germany), adenosine triphosphate (ATP) contents and microbial activities were measured in haplic Luviosls. As indices for microbial available substrate, water soluble organic carbon compounds in soils were determined and decomposable young soil organic matter was calculated from organic fertilizers and crop and root residues using empirical decomposition functions. Higher ATP contents and microbial activities were observed along with organic fertilization (liquid cattle manure) than with mineral fertilization. Shallow cultivation with a rotary cultivator led to higher values of microbial properties in the upper part of the Ap horizon than ploughing. Soil microbial parameters were higher in plots under a rape-cereals crop rotation, compared to a legumes-cereals crop rotation. Microbial biomass and its activities were related more closely to decomposable young soil organic matter than to soil humus content or to any other soil property. Water soluble organic carbon compounds did not prove as an indicator of microbial available substrate. [Spanish] La biomasa y la actividad microbianas son indicadores sensibles de los efectos del manejo del suelo. Por ejemplo, con la rotacion de cultivos se obtiene un contenido y una actividad mayores de la biomasa microbiana en contraste con el simple

  8. Microbial Production of Malic Acid from Biofuel-Related Coproducts and Biomass

    Directory of Open Access Journals (Sweden)

    Thomas P. West

    2017-04-01

    Full Text Available The dicarboxylic acid malic acid synthesized as part of the tricarboxylic acid cycle can be produced in excess by certain microorganisms. Although malic acid is produced industrially to a lesser extent than citric acid, malic acid has industrial applications in foods and pharmaceuticals as an acidulant among other uses. Only recently has the production of this organic acid from coproducts of industrial bioprocessing been investigated. It has been shown that malic acid can be synthesized by microbes from coproducts generated during biofuel production. More specifically, malic acid has been shown to be synthesized by species of the fungus Aspergillus on thin stillage, a coproduct from corn-based ethanol production, and on crude glycerol, a coproduct from biodiesel production. In addition, the fungus Ustilago trichophora has also been shown to produce malic acid from crude glycerol. With respect to bacteria, a strain of the thermophilic actinobacterium Thermobifida fusca has been shown to produce malic acid from cellulose and treated lignocellulosic biomass. An alternate method of producing malic acid is to use agricultural biomass converted to syngas or biooil as a substrate for fungal bioconversion. Production of poly(β-l-malic acid by strains of Aureobasidium pullulans from agricultural biomass has been reported where the polymalic acid is subsequently hydrolyzed to malic acid. This review examines applications of malic acid, metabolic pathways that synthesize malic acid and microbial malic acid production from biofuel-related coproducts, lignocellulosic biomass and poly(β-l-malic acid.

  9. Metabolic enzyme cost explains variable trade-offs between microbial growth rate and yield.

    Directory of Open Access Journals (Sweden)

    Meike T Wortel

    2018-02-01

    Full Text Available Microbes may maximize the number of daughter cells per time or per amount of nutrients consumed. These two strategies correspond, respectively, to the use of enzyme-efficient or substrate-efficient metabolic pathways. In reality, fast growth is often associated with wasteful, yield-inefficient metabolism, and a general thermodynamic trade-off between growth rate and biomass yield has been proposed to explain this. We studied growth rate/yield trade-offs by using a novel modeling framework, Enzyme-Flux Cost Minimization (EFCM and by assuming that the growth rate depends directly on the enzyme investment per rate of biomass production. In a comprehensive mathematical model of core metabolism in E. coli, we screened all elementary flux modes leading to cell synthesis, characterized them by the growth rates and yields they provide, and studied the shape of the resulting rate/yield Pareto front. By varying the model parameters, we found that the rate/yield trade-off is not universal, but depends on metabolic kinetics and environmental conditions. A prominent trade-off emerges under oxygen-limited growth, where yield-inefficient pathways support a 2-to-3 times higher growth rate than yield-efficient pathways. EFCM can be widely used to predict optimal metabolic states and growth rates under varying nutrient levels, perturbations of enzyme parameters, and single or multiple gene knockouts.

  10. Winter climate change affects growing-season soil microbial biomass and activity in northern hardwood forests

    Science.gov (United States)

    Jorge Durán; Jennifer L. Morse; Peter M. Groffman; John L. Campbell; Lynn M. Christenson; Charles T. Driscoll; Timothy J. Fahey; Melany C. Fisk; Myron J. Mitchell; Pamela H. Templer

    2014-01-01

    Understanding the responses of terrestrial ecosystems to global change remains a major challenge of ecological research. We exploited a natural elevation gradient in a northern hardwood forest to determine how reductions in snow accumulation, expected with climate change, directly affect dynamics of soil winter frost, and indirectly soil microbial biomass and activity...

  11. Microbial enzymes for the recycling of recalcitrant petroleum-based plastics: how far are we?

    Science.gov (United States)

    Wei, Ren; Zimmermann, Wolfgang

    2017-11-01

    Petroleum-based plastics have replaced many natural materials in their former applications. With their excellent properties, they have found widespread uses in almost every area of human life. However, the high recalcitrance of many synthetic plastics results in their long persistence in the environment, and the growing amount of plastic waste ending up in landfills and in the oceans has become a global concern. In recent years, a number of microbial enzymes capable of modifying or degrading recalcitrant synthetic polymers have been identified. They are emerging as candidates for the development of biocatalytic plastic recycling processes, by which valuable raw materials can be recovered in an environmentally sustainable way. This review is focused on microbial biocatalysts involved in the degradation of the synthetic plastics polyethylene, polystyrene, polyurethane and polyethylene terephthalate (PET). Recent progress in the application of polyester hydrolases for the recovery of PET building blocks and challenges for the application of these enzymes in alternative plastic waste recycling processes will be discussed. © 2017 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

  12. Sediment microbial activity and its relation to environmental variables along the eastern Gulf of Finland coastline

    Science.gov (United States)

    Polyak, Yulia; Shigaeva, Tatyana; Gubelit, Yulia; Bakina, Ludmila; Kudryavtseva, Valentina; Polyak, Mark

    2017-07-01

    Sediment microbial activity and its relationship with the main environmental factors and pollutants were examined in the coastal area of the eastern Gulf of Finland, Baltic Sea. The activity of two common oxidoreductase enzymes: dehydrogenase (DA) and catalase (CA) varied significantly between 13 study sites. In the Neva Bay the highest microbial activities (DA: 2.64 mg TFF (10 g- 1) day- 1, CA: 6.29 mg H2O2 g- 1) were recorded, while in the outer estuary the minimum values of dehydrogenase and catalase were measured. DA, CA, and abundances of culturable heterotrophic bacteria (CHB) were positively correlated with each other, while biomass of green opportunistic algae was independent of both microbial activities and CHB. Enzymatic activity was found to be strongly positively correlated with sediment particle size and organic matter content, but unrelated to the other studied environmental parameters (temperature, pH, and salinity). Principal components analysis (PCA), controlling for environmental variables, supported direct effects of metal and oil contamination on sediment microbial activity. Also it had shown the similar patterns for algal biomass and metals. Our results suggest that copper and hydrocarbons are the main anthropogenic variables influencing enzyme distribution along the eastern Gulf of Finland coastline.

  13. Metagenomic insights into the rumen microbial fibrolytic enzymes in Indian crossbred cattle fed finger millet straw.

    Science.gov (United States)

    Jose, V Lyju; Appoothy, Thulasi; More, Ravi P; Arun, A Sha

    2017-12-01

    The rumen is a unique natural habitat, exhibiting an unparalleled genetic resource of fibrolytic enzymes of microbial origin that degrade plant polysaccharides. The objectives of this study were to identify the principal plant cell wall-degrading enzymes and the taxonomic profile of rumen microbial communities that are associated with it. The cattle rumen microflora and the carbohydrate-active enzymes were functionally classified through a whole metagenomic sequencing approach. Analysis of the assembled sequences by the Carbohydrate-active enzyme analysis Toolkit identified the candidate genes encoding fibrolytic enzymes belonging to different classes of glycoside hydrolases(11,010 contigs), glycosyltransferases (6366 contigs), carbohydrate esterases (4945 contigs), carbohydrate-binding modules (1975 contigs), polysaccharide lyases (480 contigs), and auxiliary activities (115 contigs). Phylogenetic analysis of CAZyme encoding contigs revealed that a significant proportion of CAZymes were contributed by bacteria belonging to genera Prevotella, Bacteroides, Fibrobacter, Clostridium, and Ruminococcus. The results indicated that the cattle rumen microbiome and the CAZymes are highly complex, structurally similar but compositionally distinct from other ruminants. The unique characteristics of rumen microbiota and the enzymes produced by resident microbes provide opportunities to improve the feed conversion efficiency in ruminants and serve as a reservoir of industrially important enzymes for cellulosic biofuel production.

  14. Soil microbial activities and its relationship with soil chemical ...

    African Journals Online (AJOL)

    The fields assessed are organically managed Soils (OMS), Inorganically Managed Soils (IMS) and an Uncultivated Land having grass coverage (ULS). Soil Microbial Respiration (SMR), Microbial Biomass Carbon (MBC), Microbial Biomass Nitrogen (MBN) and Microbial Biomass Phosphorus (MBP) were analyzed.

  15. Alkaline peroxide pretreatment of corn stover: effects of biomass, peroxide, and enzyme loading and composition on yields of glucose and xylose

    Directory of Open Access Journals (Sweden)

    Hodge David B

    2011-06-01

    Full Text Available Abstract Background Pretreatment is a critical step in the conversion of lignocellulose to fermentable sugars. Although many pretreatment processes are currently under investigation, none of them are entirely satisfactory in regard to effectiveness, cost, or environmental impact. The use of hydrogen peroxide at pH 11.5 (alkaline hydrogen peroxide (AHP was shown by Gould and coworkers to be an effective pretreatment of grass stovers and other plant materials in the context of animal nutrition and ethanol production. Our earlier experiments indicated that AHP performed well when compared against two other alkaline pretreatments. Here, we explored several key parameters to test the potential of AHP for further improvement relevant to lignocellulosic ethanol production. Results The effects of biomass loading, hydrogen peroxide loading, residence time, and pH control were tested in combination with subsequent digestion with a commercial enzyme preparation, optimized mixtures of four commercial enzymes, or optimized synthetic mixtures of pure enzymes. AHP pretreatment was performed at room temperature (23°C and atmospheric pressure, and after AHP pretreatment the biomass was neutralized with HCl but not washed before enzyme digestion. Standard enzyme digestion conditions were 0.2% glucan loading, 15 mg protein/g glucan, and 48 h digestion at 50°C. Higher pretreatment biomass loadings (10% to 20% gave higher monomeric glucose (Glc and xylose (Xyl yields than the 2% loading used in earlier studies. An H2O2 loading of 0.25 g/g biomass was almost as effective as 0.5 g/g, but 0.125 g/g was significantly less effective. Optimized mixtures of four commercial enzymes substantially increased post-AHP-pretreatment enzymatic hydrolysis yields at all H2O2 concentrations compared to any single commercial enzyme. At a pretreatment biomass loading of 10% and an H2O2 loading of 0.5 g/g biomass, an optimized commercial mixture at total protein loadings of 8 or 15 mg

  16. Seasonal and diel variability in dissolved DNA and in microbial biomass and activity in a subtropical estuary

    International Nuclear Information System (INIS)

    Paul, J.H.; Deflaun, M.F.; Jeffrey, W.H.; David, A.W.

    1988-01-01

    Dissolved DNA and microbial biomass and activity parameters were measured over a 15-month period at three stations along a salinity gradient in Tampa Bay, Fla. Dissolved DNA showed seasonal variation, with minimal values in December and January and maximal values in summer months (July and August). This pattern of seasonal variation followed that of particulate DNA and water temperature and did not correlate with bacterioplankton (direct counts and [ 3 H] thymidine incorporation) or phytoplankton (chlorophyll α and 14 CO 2 fixation) biomass and activity. Microautotrophic populations showed maxima in the spring and fall, whereas microheterotrophic activity was greatest in late summer (September). Both autotrophic and heterotrophic microbial activity was greatest at the high estuarine (low salinity) station and lowest at the mouth of the bay (high salinity station), irrespective of season. Dissolved DNA carbon and phosphorus constituted 0.11 +/- 0.05% of the dissolved organic carbon and 6.6 +/- 6.5% of the dissolved organic phosphorus, respectively. Strong diel periodicity was noted in dissolved DNA and in microbial activity in Bayboro Harbor during the dry season. A noon maximum in primary productivity was followed by an 8 p.m. maximum in heterotrophic activity and a midnight maximum in dissolved DNA. This diel periodicity was less pronounced in the wet season, when microbial parameters were strongly influenced by episodic inputs of freshwater

  17. A stepwise-cluster microbial biomass inference model in food waste composting

    International Nuclear Information System (INIS)

    Sun Wei; Huang, Guo H.; Zeng Guangming; Qin Xiaosheng; Sun Xueling

    2009-01-01

    A stepwise-cluster microbial biomass inference (SMI) model was developed through introducing stepwise-cluster analysis (SCA) into composting process modeling to tackle the nonlinear relationships among state variables and microbial activities. The essence of SCA is to form a classification tree based on a series of cutting or mergence processes according to given statistical criteria. Eight runs of designed experiments in bench-scale reactors in a laboratory were constructed to demonstrate the feasibility of the proposed method. The results indicated that SMI could help establish a statistical relationship between state variables and composting microbial characteristics, where discrete and nonlinear complexities exist. Significance levels of cutting/merging were provided such that the accuracies of the developed forecasting trees were controllable. Through an attempted definition of input effects on the output in SMI, the effects of the state variables on thermophilic bacteria were ranged in a descending order as: Time (day) > moisture content (%) > ash content (%, dry) > Lower Temperature (deg. C) > pH > NH 4 + -N (mg/Kg, dry) > Total N (%, dry) > Total C (%, dry); the effects on mesophilic bacteria were ordered as: Time > Upper Temperature (deg. C) > Total N > moisture content > NH 4 + -N > Total C > pH. This study made the first attempt in applying SCA to mapping the nonlinear and discrete relationships in composting processes.

  18. Microbial communities in litter and soil - particles size fractionation, C- and N-pools and soil enzymes

    International Nuclear Information System (INIS)

    Stemmer, M.; Gerzabek, M.H.; Pichlmayer, F.; Kandeler, E.

    1995-08-01

    In this study we try to correlate C and N pool investigations to enzyme activities in particle size fractions of soils. Soil incubations in the lab (for one year) simulate two different conventional tillage treatments : (i) soil mixed with maize straw (GSF-mixed) and (ii) soil with maize straw lying on the top (home-mixed). The control soil is incubated without any amendment. The separation of the particle size fractions (2000 - 200 μm, 200 - 63 μm, 63 - 2 μm, 2 - 0.1 μm and 0.1 - 0 μm) is realized by a combination of wet-sieving and centrifugation. To disrupt aggregates we use a defined low-energy ultrasonication, which partly preserves microaggregates. The decomposition of organic C during the incubation can be observed clearly, the small amount of N in the added maize straw complicates the analysis. The isotopic measurements of δ13C and δ15N provide valuable additional informations in this context. Both enzymes, saccharase and xylanase, seem to react in a more sensitive way on the incorporation of the maize litter, than the chemical analysis of the pools. The saccharase activity, which seems to be a sensitive indicator for microbial biomass, shows different behaviour between the mix- and top-treatment. The xylanase activity is mainly located in the coarse sand fraction, this extracellular enzyme might be adsorbed by the particulate organic matter. The transfer of adhering coatings and small particles of the added maize to small sized particles during the fractionation procedure and the 'passive role' of the silt fraction, which could be due to the used method, are nonexpected results. (author)

  19. Optimizing Cofactor Specificity of Oxidoreductase Enzymes for the Generation of Microbial Production Strains—OptSwap

    DEFF Research Database (Denmark)

    King, Zachary A.; Feist, Adam

    2013-01-01

    Central oxidoreductase enzymes (eg, dehydrogenases, reductases) in microbial metabolism often have preferential binding specificity for one of the two major currency metabolites NAD(H) and NADP(H). These enzyme specificities result in a division of the metabolic functionality of the currency...... specificities of oxidoreductase enzyme and complementary reaction knockouts. Using the Escherichia coli genome-scale metabolic model iJO1366, OptSwap predicted eight growth-coupled production designs with significantly greater product yields or substrate-specific productivities than designs predicted with gene...

  20. Combined effects of reduced irrigation and water quality on the soil microbial community of a citrus orchard under semi-arid conditions

    Czech Academy of Sciences Publication Activity Database

    Bastida, F.; Torres, I.F.; Romero-Trigueros, C.; Baldrian, Petr; Větrovský, Tomáš; Bayona, J.M.; Alarcón, J.J.; Hernandéz, T.; García, C.; Nikolás, E.

    2017-01-01

    Roč. 104, JAN 2017 (2017), s. 226-237 ISSN 0038-0717 Institutional support: RVO:61388971 Keywords : Microbial biomass * Bacterial diversity * Enzyme activities Subject RIV: EE - Microbiology, Virology OBOR OECD: Microbiology Impact factor: 4.857, year: 2016

  1. Production of L-lactic Acid from Biomass Wastes Using Scallop Crude Enzymes and Novel Lactic Acid Bacterium

    Science.gov (United States)

    Yanagisawa, Mitsunori; Nakamura, Kanami; Nakasaki, Kiyohiko

    In the present study, biomass waste raw materials including paper mill sludge, bamboo, sea lettuce, and shochu residue (from a distiller) and crude enzymes derived from inedible and discarded scallop parts were used to produce L-lactic acid for the raw material of biodegradable plastic poly-lactic acid. The activities of cellulase and amylase in the crude enzymes were 22 and 170units/L, respectively, and L-lactic acid was produced from every of the above mentioned biomass wastes, by the method of liquid-state simultaneous saccharification and fermentation (SSF) . The L-lactic acid concentrations produced from sea lettuce and shochu residue, which contain high concentration of starch were 3.6 and 9.3g/L, respectively, and corresponded to greater than 25% of the conversion of glucans contained in these biomass wastes. Furthermore, using the solid state SSF method, concentrations as high as 13g/L of L-lactic acid were obtained from sea lettuce and 26g/L were obtained from shochu residue.

  2. Microbial decomposers not constrained by climate history along a Mediterranean climate gradient in southern California.

    Science.gov (United States)

    Baker, Nameer R; Khalili, Banafshe; Martiny, Jennifer B H; Allison, Steven D

    2018-06-01

    Microbial decomposers mediate the return of CO 2 to the atmosphere by producing extracellular enzymes to degrade complex plant polymers, making plant carbon available for metabolism. Determining if and how these decomposer communities are constrained in their ability to degrade plant litter is necessary for predicting how carbon cycling will be affected by future climate change. We analyzed mass loss, litter chemistry, microbial biomass, extracellular enzyme activities, and enzyme temperature sensitivities in grassland litter transplanted along a Mediterranean climate gradient in southern California. Microbial community composition was manipulated by caging litter within bags made of nylon membrane that prevent microbial immigration. To test whether grassland microbes were constrained by climate history, half of the bags were inoculated with local microbial communities native to each gradient site. We determined that temperature and precipitation likely interact to limit microbial decomposition in the extreme sites along our gradient. Despite their unique climate history, grassland microbial communities were not restricted in their ability to decompose litter under different climate conditions across the gradient, although microbial communities across our gradient may be restricted in their ability to degrade different types of litter. We did find some evidence that local microbial communities were optimized based on climate, but local microbial taxa that proliferated after inoculation into litterbags did not enhance litter decomposition. Our results suggest that microbial community composition does not constrain C-cycling rates under climate change in our system, but optimization to particular resource environments may act as more general constraints on microbial communities. © 2018 by the Ecological Society of America.

  3. Microbial degradation of coconut coir dust for biomass production

    Energy Technology Data Exchange (ETDEWEB)

    Uyenco, F.R.; Ochoa, J.A.K.

    Several species of white-rot fungi were studied for its ability to degrade the lignocellulose components of coir dust at optimum conditions. The most effective fungi was Phanerochaeta chrysosporium UPCC 4003. This organism degraded the lignocellulose complex of coir dust at a rate of about 25 percent in 4 weeks. The degradation process was carried on with minimal nitrogen concentration, coconut water supplementation and moisture levels between 85-90 percent. Shake flask cultures of the degraded coir dust using cellulolytic fungi were not effective. In fermentor cultures with Chaetomium cellulolyticum UPCC 3934, supplemented coir dust was converted into a microbial biomass product (MBP) with 15.58 percent lignin, 19.20 percent cellulose and 18.87 percent protein. More work is being done on the utilization of coir dust on a low technology.

  4. Structural stability, microbial biomass and community composition of sediments affected by the hydric dynamics of an urban stormwater infiltration basin. Dynamics of physical and microbial characteristics of stormwater sediment.

    Science.gov (United States)

    Badin, Anne Laure; Monier, Armelle; Volatier, Laurence; Geremia, Roberto A; Delolme, Cécile; Bedell, Jean-Philippe

    2011-05-01

    The sedimentary layer deposited at the surface of stormwater infiltration basins is highly organic and multicontaminated. It undergoes considerable moisture content fluctuations due to the drying and inundation cycles (called hydric dynamics) of these basins. Little is known about the microflora of the sediments and its dynamics; hence, the purpose of this study is to describe the physicochemical and biological characteristics of the sediments at different hydric statuses of the infiltration basin. Sediments were sampled at five time points following rain events and dry periods. They were characterized by physical (aggregation), chemical (nutrients and heavy metals), and biological (total, bacterial and fungal biomasses, and genotypic fingerprints of total bacterial and fungal communities) parameters. Data were processed using statistical analyses which indicated that heavy metal (1,841 μg/g dry weight (DW)) and organic matter (11%) remained stable through time. By contrast, aggregation, nutrient content (NH₄⁺, 53-717 μg/g DW), pH (6.9-7.4), and biological parameters were shown to vary with sediment water content and sediment biomass, and were higher consecutive to stormwater flows into the basin (up to 7 mg C/g DW) than during dry periods (0.6 mg C/g DW). Coinertia analysis revealed that the structure of the bacterial communities is driven by the hydric dynamics of the infiltration basin, although no such trend was found for fungal communities. Hydric dynamics more than rain events appear to be more relevant for explaining variations of aggregation, microbial biomass, and shift in the microbial community composition. We concluded that the hydric dynamics of stormwater infiltration basins greatly affects the structural stability of the sedimentary layer, the biomass of the microbial community living in it and its dynamics. The decrease in aggregation consecutive to rewetting probably enhances access to organic matter (OM), explaining the consecutive release

  5. Planktonic food web structure at a coastal time-series site: I. Partitioning of microbial abundances and carbon biomass

    Science.gov (United States)

    Caron, David A.; Connell, Paige E.; Schaffner, Rebecca A.; Schnetzer, Astrid; Fuhrman, Jed A.; Countway, Peter D.; Kim, Diane Y.

    2017-03-01

    Biogeochemistry in marine plankton communities is strongly influenced by the activities of microbial species. Understanding the composition and dynamics of these assemblages is essential for modeling emergent community-level processes, yet few studies have examined all of the biological assemblages present in the plankton, and benchmark data of this sort from time-series studies are rare. Abundance and biomass of the entire microbial assemblage and mesozooplankton (>200 μm) were determined vertically, monthly and seasonally over a 3-year period at a coastal time-series station in the San Pedro Basin off the southwestern coast of the USA. All compartments of the planktonic community were enumerated (viruses in the femtoplankton size range [0.02-0.2 μm], bacteria + archaea and cyanobacteria in the picoplankton size range [0.2-2.0 μm], phototrophic and heterotrophic protists in the nanoplanktonic [2-20 μm] and microplanktonic [20-200 μm] size ranges, and mesozooplankton [>200 μm]. Carbon biomass of each category was estimated using standard conversion factors. Plankton abundances varied over seven orders of magnitude across all categories, and total carbon biomass averaged approximately 60 μg C l-1 in surface waters of the 890 m water column over the study period. Bacteria + archaea comprised the single largest component of biomass (>1/3 of the total), with the sum of phototrophic protistan biomass making up a similar proportion. Temporal variability at this subtropical station was not dramatic. Monthly depth-specific and depth-integrated biomass varied 2-fold at the station, while seasonal variances were generally web structure and function at this coastal observatory.

  6. Soil-derived microbial consortia enriched with different plant biomass reveal distinct players acting in lignocellulose degradation

    NARCIS (Netherlands)

    de Lima Brossi, Maria Julia; Jiménez Avella, Diego; Cortes Tolalpa, Larisa; van Elsas, Jan

    Here, we investigated how different plant biomass, and-for one substrate-pH, drive the composition of degrader microbial consortia. We bred such consortia from forest soil, incubated along nine aerobic sequential - batch enrichments with wheat straw (WS1, pH 7.2; WS2, pH 9.0), switchgrass (SG, pH

  7. Soil microbial biomass, activity and community composition along altitudinal gradients in the High Arctic (Billefjorden, Svalbard)

    Czech Academy of Sciences Publication Activity Database

    Kotas, P.; Šantrůčková, H.; Elster, Josef; Kaštovská, E.

    2018-01-01

    Roč. 15, č. 6 (2018), s. 1879-1894 ISSN 1726-4170 R&D Projects: GA MŠk(CZ) LM2015075 Grant - others:GA MŠk LM2010009 Institutional support: RVO:67985939 Keywords : ecosystem * High Arctic * soil microbial biomass Subject RIV: EH - Ecology, Behaviour OBOR OECD: Ecology Impact factor: 3.851, year: 2016

  8. Microbial Community Structure of an Alluvial Aquifer Treated to Encourage Microbial Induced Calcite Precipitation

    Science.gov (United States)

    Ohan, J.; Saneiyan, S.; Lee, J.; Ntarlagiannis, D.; Burns, S.; Colwell, F. S.

    2017-12-01

    An oligotrophic aquifer in the Colorado River floodplain (Rifle, CO) was treated with molasses and urea to encourage microbial induced calcite precipitation (MICP). This would stabilize the soil mass by reducing porosity and strengthening the mineral fabric. Over the course of a 15-day treatment period, microbial biomass was collected from monitoring well groundwater for DNA extraction and sequencing. Bromide, a conservative tracer, was co-injected and subsequently detected in downgradient wells, confirming effective nutrient delivery. Conductivity increased during the injection regime and an overall decrease in pH was observed. Groundwater chemistry showed a marked increase in ammonia, suggesting urea hydrolysis - a process catalyzed by the enzyme urease - the primary enzyme implicated in MICP. Additionally, soluble iron was detected, suggesting a general increase in microbial activity; possibly as iron-reducing bacteria changed insoluble ferric oxide to soluble ferrous hydroxide in the anoxic aquifer. DNA sequencing of the 16S rRNA gene confirmed the presence of iron reducing bacteria, including Shewanella and Desulfuromonadales. Generally, a decrease in microbial community diversity was observed when pre-injection community taxa were compared with post-injection community taxa. Phyla indicative of anoxic aquifers were represented in accordance with previous literature at the Rifle site. Linear discriminant analysis showed significant differences in representative phyla over the course of the injection series. Geophysical monitoring of the site further suggested changes that could be due to MICP. Induced polarization increased the phase shift in the primary treated area, in agreement with laboratory experiments. Cross-hole seismic testing confirmed that the shear wave velocities increased in the treated soil mass, implying the soil matrix became more stable. Future investigations will help elucidate the viability and efficacy of MICP treatment in changing

  9. Microbial Energy Conversion

    Energy Technology Data Exchange (ETDEWEB)

    Buckley, Merry [American Society for Microbiology (ASM), Washington, DC (United States); Wall, Judy D. [Univ. of Missouri, Columbia, MO (United States)

    2006-10-01

    natural gas from the subsurface. The participants discussed--key microbial conversion paths; overarching research issues; current funding models and microbial energy research; education, training, interdisciplinary cooperation and communication. Their recommendations are--Cellulose and lignocellulose are the preferred substrates for producing liquid transportation fuels, of which ethanol is the most commonly considered example. Generating fuels from these materials is still difficult and costly. A number of challenges need to be met in order to make the conversion of cellulose and lignocellulose to transportation fuels more cost-competitive. The design of hydrogen-producing bioreactors must be improved in order to more effectively manage hydrogen removal, oxygen exclusion, and, in the case of photobioreactors, to capture light energy more efficiently. Methane production may be optimized by fine-tuning methanogenic microbial communities. The ability to transfer electrons to an anode in a microbial fuel cell is probably very broadly distributed in the bacterial world. The scientific community needs a larger inventory of cultivated microorganisms from which to draw for energy conversion development. New and unusual organisms for manufacturing fuels and for use in fuel cells can be discovered using bioprospecting techniques. Particular emphasis should be placed on finding microbes, microbial communities, and enzymes that can enhance the conversion of lignocellulosic biomass to usable sugars. Many of the microbial processes critical to energy conversion are carried out by complex communities of organisms, and there is a need to better understand the community interactions that make these transformations possible. Better understanding of microbial community structure, robustness, networks, homeostasis, and cell-to-cell signaling is also needed. A better understanding of the basic enzymology of microorganisms is needed in order to move forward more quickly with microbial energy

  10. Responses of absolute and specific enzyme activity to consecutive application of composted sewage sludge in a Fluventic Ustochrept.

    Directory of Open Access Journals (Sweden)

    Xiao Liu

    Full Text Available Composted sewage sludge (CS is considered a rich source of soil nutrients and significantly affects the physical, chemical, and biological characteristics of soil, but its effect on specific enzyme activity in soil is disregarded. The present experiment examined the absolute and specific enzyme activity of the enzymes involved in carbon, nitrogen, and phosphorus cycles, the diversity of soil microbial functions, and soil community composition in a Fluventic Ustochrept under a maize-wheat rotation system in North China during 2012-2015. Application of CS led to increase in MBC and in its ratio to both total organic carbon (TOC and microbial biomass nitrogen (MBN. Absolute enzyme activity, except that of phosphatase, increased in CS-treated soils, whereas specific activity of all the enzymes declined, especially at the highest dose of CS (45 t ha-1. The diversity of soil microbial community also increased in CS-treated soils, whereas its functional diversity declined at higher doses of CS owing to the lowered specific enzyme activity. These changes indicate that CS application induced the domination of microorganisms that are not metabolically active and those that use resources more efficiently, namely fungi. Redundancy analysis showed that fundamental alterations in soil enzyme activity depend on soil pH. Soil specific enzyme activity is affected more than absolute enzyme activity by changes in soil properties, especially soil microbial activity and composition of soil microflora (as judged by the following ratios: MBC/TOC, MBC/MBN, and TOC/LOC, that is labile organic carbon through the Pearson Correlation Coefficient. Specific enzyme activity is thus a more accurate parameter than absolute enzyme activity for monitoring the effect of adding CS on the activities and structure of soil microbial community.

  11. Responses of microbial biomass carbon and nitrogen to experimental warming: a meta-analysis

    Science.gov (United States)

    Xu, W.; Yuan, W.

    2017-12-01

    Soil microbes play important roles in regulating terrestrial carbon and nitrogen cycling and strongly influence feedbacks of ecosystem to global warming. However, the inconsistent responses of microbial biomass carbon (MBC) and nitrogen (MBN) to experimental warming have been observed, and the response on ratio between MBC and MBN (MBC:MBN) has not been identified. This meta-analysis synthesized the warming experiments at 58 sites globally to investigate the responses of MBC:MBN to climate warming. Our results showed that warming significantly increased MBC by 3.61 ± 0.80% and MBN by 5.85 ± 0.90% and thus decreased the MBC:MBN by 3.34 ± 0.66%. MBC showed positive responses to warming but MBN exhibited negative responses to warming at low warming magnitude (2°C) the results were inverted. The different effects of warming magnitude on microbial biomass resulted from the warming-induced decline in soil moisture and substrate supply. Moreover, MBC and MBN had strong positive responses to warming at the mid-term (3-4 years) or short-term (1-2 years) duration, but the responses tended to decrease at long-term (≥ 5 years) warming duration. This study fills the knowledge gap on the responses of MBC:MBN to warming and may benefit the development of coupled carbon and nitrogen models.

  12. Microbial activities in forest soils exposed to chronic depositions from a lignite power plant

    Energy Technology Data Exchange (ETDEWEB)

    Klose, S.; Wernecke, K.D.; Makeschin, F. [Technical University of Dresden, Tharandt (Germany)

    2004-12-01

    Atmospheric emissions of fly ash and SO{sub 2} from lignite-fired power plants strongly affect large forest areas in Germany. The impact of different deposition loads on the microbial biomass and enzyme activities was studied at three forest sites (Picea abies (L.) Karst.) along an emission gradient of 3, 6, and 15 km downwind of a coal-fired power plant, representing high, moderate and low emission rates. An additional site at a distance of 3 km from the power plant was chosen to study the influence of forest type on microbial parameters in coniferous forest soils under fly ash and SO{sub 2} emissions. Soil microbial biomass C and N, CO{sub 2} evolved and activities of L-asparaginase, L-glutaminase, beta-glucosidase, acid phosphatase and arylsulfatase (expressed on dry soil and organic C basis) were determined in the forest floor (L, Of and Oh horizon) and mineral top soil (0-10 cm). It is concluded that chronic fly ash depositions decrease litter decomposition by influencing specific microbial and enzymatic processes in forest soils.

  13. Microbial conversion of biomass into bio-based polymers.

    Science.gov (United States)

    Kawaguchi, Hideo; Ogino, Chiaki; Kondo, Akihiko

    2017-12-01

    The worldwide market for plastics is rapidly growing, and plastics polymers are typically produced from petroleum-based chemicals. The overdependence on petroleum-based chemicals for polymer production raises economic and environmental sustainability concerns. Recent progress in metabolic engineering has expanded fermentation products from existing aliphatic acids or alcohols to include aromatic compounds. This diversity provides an opportunity to expand the development and industrial uses of high-performance bio-based polymers. However, most of the biomonomers are produced from edible sugars or starches that compete directly with food and feed uses. The present review focuses on recent progress in the microbial conversion of biomass into bio-based polymers, in which fermentative products from renewable feedstocks serve as biomonomers for the synthesis of bio-based polymers. In particular, the production of biomonomers from inedible lignocellulosic feedstocks by metabolically engineered microorganisms and the synthesis of bio-based engineered plastics from the biological resources are discussed. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Effects of microbial enzymes on starch and hemicellulose degradation in total mixed ration silages

    Directory of Open Access Journals (Sweden)

    Tingting Ning

    2017-02-01

    Full Text Available Objective This study investigated the association of enzyme-producing microbes and their enzymes with starch and hemicellulose degradation during fermentation of total mixed ration (TMR silage. Methods The TMRs were prepared with soybean curd residue, alfalfa hay (ATMR or Leymus chinensis hay (LTMR, corn meal, soybean meal, vitamin-mineral supplements, and salt at a ratio of 25:40:30:4:0.5:0.5 on a dry matter basis. Laboratory-scale bag silos were randomly opened after 1, 3, 7, 14, 28, and 56 days of ensiling and subjected to analyses of fermentation quality, carbohydrates loss, microbial amylase and hemicellulase activities, succession of dominant amylolytic or hemicellulolytic microbes, and their microbial and enzymatic properties. Results Both ATMR and LTMR silages were well preserved, with low pH and high lactic acid concentrations. In addition to the substantial loss of water soluble carbohydrates, loss of starch and hemicellulose was also observed in both TMR silages with prolonged ensiling. The microbial amylase activity remained detectable throughout the ensiling in both TMR silages, whereas the microbial hemicellulase activity progressively decreased until it was inactive at day 14 post-ensiling in both TMR silages. During the early stage of fermentation, the main amylase-producing microbes were Bacillus amyloliquefaciens (B. amyloliquefaciens, B. cereus, B. licheniformis, and B. subtilis in ATMR silage and B. flexus, B. licheniformis, and Paenibacillus xylanexedens (P. xylanexedens in LTMR silage, whereas Enterococcus faecium was closely associated with starch hydrolysis at the later stage of fermentation in both TMR silages. B. amyloliquefaciens, B. licheniformis, and B. subtilis and B. licheniformis, B. pumilus, and P. xylanexedens were the main source of microbial hemicellulase during the early stage of fermentation in ATMR and LTMR silages, respectively. Conclusion The microbial amylase contributes to starch hydrolysis during the

  15. Community analysis of plant biomass-degrading microorganisms from Obsidian Pool, Yellowstone National Park.

    Science.gov (United States)

    Vishnivetskaya, Tatiana A; Hamilton-Brehm, Scott D; Podar, Mircea; Mosher, Jennifer J; Palumbo, Anthony V; Phelps, Tommy J; Keller, Martin; Elkins, James G

    2015-02-01

    The conversion of lignocellulosic biomass into biofuels can potentially be improved by employing robust microorganisms and enzymes that efficiently deconstruct plant polysaccharides at elevated temperatures. Many of the geothermal features of Yellowstone National Park (YNP) are surrounded by vegetation providing a source of allochthonic material to support heterotrophic microbial communities adapted to utilize plant biomass as a primary carbon and energy source. In this study, a well-known hot spring environment, Obsidian Pool (OBP), was examined for potential biomass-active microorganisms using cultivation-independent and enrichment techniques. Analysis of 33,684 archaeal and 43,784 bacterial quality-filtered 16S rRNA gene pyrosequences revealed that archaeal diversity in the main pool was higher than bacterial; however, in the vegetated area, overall bacterial diversity was significantly higher. Of notable interest was a flooded depression adjacent to OBP supporting a stand of Juncus tweedyi, a heat-tolerant rush commonly found growing near geothermal features in YNP. The microbial community from heated sediments surrounding the plants was enriched in members of the Firmicutes including potentially (hemi)cellulolytic bacteria from the genera Clostridium, Anaerobacter, Caloramator, Caldicellulosiruptor, and Thermoanaerobacter. Enrichment cultures containing model and real biomass substrates were established at a wide range of temperatures (55-85 °C). Microbial activity was observed up to 80 °C on all substrates including Avicel, xylan, switchgrass, and Populus sp. Independent of substrate, Caloramator was enriched at lower (65 °C) temperatures.

  16. Ionic liquid-tolerant microorganisms and microbial communities for lignocellulose conversion to bioproducts.

    Science.gov (United States)

    Yu, Chaowei; Simmons, Blake A; Singer, Steven W; Thelen, Michael P; VanderGheynst, Jean S

    2016-12-01

    Chemical and physical pretreatment of biomass is a critical step in the conversion of lignocellulose to biofuels and bioproducts. Ionic liquid (IL) pretreatment has attracted significant attention due to the unique ability of certain ILs to solubilize some or all components of the plant cell wall. However, these ILs inhibit not only the enzyme activities but also the growth and productivity of microorganisms used in downstream hydrolysis and fermentation processes. While pretreated biomass can be washed to remove residual IL and reduce inhibition, extensive washing is costly and not feasible in large-scale processes. IL-tolerant microorganisms and microbial communities have been discovered from environmental samples and studies begun to elucidate mechanisms of IL tolerance. The discovery of IL tolerance in environmental microbial communities and individual microbes has lead to the proposal of molecular mechanisms of resistance. In this article, we review recent progress on discovering IL-tolerant microorganisms, identifying metabolic pathways and mechanisms of tolerance, and engineering microorganisms for IL tolerance. Research in these areas will yield new approaches to overcome inhibition in lignocellulosic biomass bioconversion processes and increase opportunities for the use of ILs in biomass pretreatment.

  17. Dominant trees affect microbial community composition and activity in post-mining afforested soils

    Czech Academy of Sciences Publication Activity Database

    Šnajdr, Jaroslav; Dobiášová, Petra; Urbanová, Michaela; Petránková, Mirka; Cajthaml, Tomáš; Frouz, Jan; Baldrian, Petr

    2013-01-01

    Roč. 56, JAN 2013 (2013), s. 105-115 ISSN 0038-0717 R&D Projects: GA MŠk(CZ) OC10064; GA ČR GAP504/12/1288; GA MŠk 2B08023 Institutional support: RVO:61388971 ; RVO:60077344 Keywords : Enzyme activity * Litter * Microbial biomass Subject RIV: EE - Microbiology, Virology; EH - Ecology, Behaviour (BC-A) Impact factor: 4.410, year: 2013

  18. Characterization of three plant biomass-degrading microbial consortia by metagenomics- and metasecretomics-based approaches

    DEFF Research Database (Denmark)

    Jiménez, Diego Javier; Brossi, Maria Julia de Lima; Schückel, Julia

    2016-01-01

    ). The highest degradation rates of lignin (~59 %) were observed with SG-M, whereas CS-M showed a high consumption of cellulose and hemicellulose. Analyses of the carbohydrate-active enzymes in the three microbial consortia showed the dominance of glycosyl hydrolases (e.g. of families GH3, GH43, GH13, GH10, GH29......), switchgrass (SG-M) and corn stover (CS-M) under aerobic and mesophilic conditions. Molecular fingerprintings, bacterial 16S ribosomal RNA (rRNA) gene amplicon sequencing and metagenomic analyses showed that the three microbial consortia were taxonomically distinct. Based on the taxonomic affiliation...

  19. Optimisation of synergistic biomass-degrading enzyme systems for efficient rice straw hydrolysis using an experimental mixture design.

    Science.gov (United States)

    Suwannarangsee, Surisa; Bunterngsook, Benjarat; Arnthong, Jantima; Paemanee, Atchara; Thamchaipenet, Arinthip; Eurwilaichitr, Lily; Laosiripojana, Navadol; Champreda, Verawat

    2012-09-01

    Synergistic enzyme system for the hydrolysis of alkali-pretreated rice straw was optimised based on the synergy of crude fungal enzyme extracts with a commercial cellulase (Celluclast™). Among 13 enzyme extracts, the enzyme preparation from Aspergillus aculeatus BCC 199 exhibited the highest level of synergy with Celluclast™. This synergy was based on the complementary cellulolytic and hemicellulolytic activities of the BCC 199 enzyme extract. A mixture design was used to optimise the ternary enzyme complex based on the synergistic enzyme mixture with Bacillus subtilis expansin. Using the full cubic model, the optimal formulation of the enzyme mixture was predicted to the percentage of Celluclast™: BCC 199: expansin=41.4:37.0:21.6, which produced 769 mg reducing sugar/g biomass using 2.82 FPU/g enzymes. This work demonstrated the use of a systematic approach for the design and optimisation of a synergistic enzyme mixture of fungal enzymes and expansin for lignocellulosic degradation. Copyright © 2012 Elsevier Ltd. All rights reserved.

  20. Monitoring Soil Microbial Activities in Different Cropping Systems Using Combined Methods

    Institute of Scientific and Technical Information of China (English)

    YUAN Zhimin; LIU Haijun; HAN Jun; SUN Jingjing; WU Xiaoying; YAO Jun

    2017-01-01

    Cropping activities may affect soil microbial activities and biomass,which would affect C and N cycling in soil and thus the crop yields and quality.In the present study,a combination of microcalorimetric,enzyme activity (sucrase,urease,catalase,and fluorescein diacetate hydrolysis),and real-time polymerase chain reaction (RT-PCR) analyses was used to investigate microbial status of farmland soils,collected from 5 different sites in Huazhong Agriculture University,China.Our results showed that among the 5 sites,both positive and negative impacts of cropping activities on soil microbial activity were observed.Enzyme activity analysis showed that cropping activities reduced soil sucrase and urease activities,which would influence the C and N cycles in soil.Much more attentions should be given to microbial status affected by cropping activities in future.According to the correlation analysis,fluorescein diacetate hydrolysis showed a significantly (P < 0.05) negative correlation with the time to reach the maximum power output (R =--0.898),but a significantly (P < 0.05) positive correlation with bacterial gene copy number (R =0.817).Soil catalase activity also showed a significantly (P < 0.05) positive correlation with bacterial gene copy number (R =0.965).Using combined methods would provide virtual information of soil microbial status.

  1. Stoichiometric imbalances between terrestrial decomposer communities and their resources: mechanisms and implications of microbial adaptations to their resources

    Directory of Open Access Journals (Sweden)

    Maria eMooshammer

    2014-02-01

    Full Text Available Terrestrial microbial decomposer communities thrive on a wide range of organic matter types that rarely ever meet their elemental demands. In this review we synthesize the current state-of-the-art of microbial adaptations to resource stoichiometry, in order to gain a deeper understanding of the interactions between heterotrophic microbial communities and their chemical environment. The stoichiometric imbalance between microbial communities and their organic substrates generally decreases from wood to leaf litter and further to topsoil and subsoil organic matter. Microbial communities can respond to these imbalances in four ways: first, they adapt their biomass composition towards their resource in a non-homeostatic behaviour. Such changes are, however, only moderate, and occur mainly because of changes in microbial community structure and less so due to cellular storage of elements in excess. Second, microbial communities can mobilize resources that meet their elemental demand by producing specific extracellular enzymes, which, in turn, is restricted by the C and N requirement for enzyme production itself. Third, microbes can regulate their element use efficiencies (ratio of element invested in growth over total element uptake, such that they release elements in excess depending on their demand (e.g., respiration and N mineralization. Fourth, diazotrophic bacteria and saprotrophic fungi may trigger the input of external N and P to decomposer communities. Theoretical considerations show that adjustments in element use efficiencies may be the most important mechanism by which microbes regulate their biomass stoichiometry. This review summarizes different views on how microbes cope with imbalanced supply of C, N and P, thereby providing a framework for integrating and linking microbial adaptation to resource imbalances to ecosystem scale fluxes across scales and ecosystems.

  2. Microbial and biochemical studies on phytase enzyme in some microorganisms

    International Nuclear Information System (INIS)

    Abdelbary, N.A.

    1997-01-01

    Mixed calcium and magnesium salts of phytic acid myoinositol hexa phosphoric acid are widely distributed in food stuffs of plant origin, they may bind essential proteins, phospholipids and microelements to form indigestible compounds. In this concern, destruction of phytic acid and its salts by different methods is very important, one of them is by using microbial phytase. This study aims to produce phytase enzyme from microorganisms and study the best conditions of production and purification and also the properties of the partially purified phytase. 22 figs., 29 tabs., 61 refs

  3. Cellulolytic and xylanolytic potential of high β-glucosidase-producing Trichoderma from decaying biomass.

    Science.gov (United States)

    Okeke, Benedict C

    2014-10-01

    Availability, cost, and efficiency of microbial enzymes for lignocellulose bioconversion are central to sustainable biomass ethanol technology. Fungi enriched from decaying biomass and surface soil mixture displayed an array of strong cellulolytic and xylanolytic activities. Strains SG2 and SG4 produced a promising array of cellulolytic and xylanolytic enzymes including β-glucosidase, usually low in cultures of Trichoderma species. Nucleotide sequence analysis of internal transcribed spacer 2 (ITS2) region of rRNA gene revealed that strains SG2 and SG4 are closely related to Trichoderma inhamatum, Trichoderma piluliferum, and Trichoderma aureoviride. Trichoderma sp. SG2 crude culture supernatant correspondingly displayed as much as 9.84 ± 1.12, 48.02 ± 2.53, and 30.10 ± 1.11 units mL(-1) of cellulase, xylanase, and β-glucosidase in 30 min assay. Ten times dilution of culture supernatant of strain SG2 revealed that total activities were about 5.34, 8.45, and 2.05 orders of magnitude higher than observed in crude culture filtrate for cellulase, xylanase, and β-glucosidase, respectively, indicating that more enzymes are present to contact with substrates in biomass saccharification. In parallel experiments, Trichoderma species SG2 and SG4 produced more β-glucosidase than the industrial strain Trichoderma reesei RUT-C30. Results indicate that strains SG2 and SG4 have potential for low cost in-house production of primary lignocellulose-hydrolyzing enzymes for production of biomass saccharides and biofuel in the field.

  4. Microbial community structure and activity in trace element-contaminated soils phytomanaged by Gentle Remediation Options (GRO).

    Science.gov (United States)

    Touceda-González, M; Prieto-Fernández, Á; Renella, G; Giagnoni, L; Sessitsch, A; Brader, G; Kumpiene, J; Dimitriou, I; Eriksson, J; Friesl-Hanl, W; Galazka, R; Janssen, J; Mench, M; Müller, I; Neu, S; Puschenreiter, M; Siebielec, G; Vangronsveld, J; Kidd, P S

    2017-12-01

    Gentle remediation options (GRO) are based on the combined use of plants, associated microorganisms and soil amendments, which can potentially restore soil functions and quality. We studied the effects of three GRO (aided-phytostabilisation, in situ stabilisation and phytoexclusion, and aided-phytoextraction) on the soil microbial biomass and respiration, the activities of hydrolase enzymes involved in the biogeochemical cycles of C, N, P, and S, and bacterial community structure of trace element contaminated soils (TECS) from six field trials across Europe. Community structure was studied using denaturing gradient gel electrophoresis (DGGE) fingerprinting of Bacteria, α- and β-Proteobacteria, Actinobacteria and Streptomycetaceae, and sequencing of DGGE bands characteristic of specific treatments. The number of copies of genes involved in ammonia oxidation and denitrification were determined by qPCR. Phytomanagement increased soil microbial biomass at three sites and respiration at the Biogeco site (France). Enzyme activities were consistently higher in treated soils compared to untreated soils at the Biogeco site. At this site, microbial biomass increased from 696 to 2352 mg ATP kg -1 soil, respiration increased from 7.4 to 40.1 mg C-CO 2 kg -1 soil d -1 , and enzyme activities were 2-11-fold higher in treated soils compared to untreated soil. Phytomanagement induced shifts in the bacterial community structure at both, the total community and functional group levels, and generally increased the number of copies of genes involved in the N cycle (nirK, nirS, nosZ, and amoA). The influence of the main soil physico-chemical properties and trace element availability were assessed and eventual site-specific effects elucidated. Overall, our results demonstrate that phytomanagement of TECS influences soil biological activity in the long term. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Effects of fire on soil nitrogen dynamics and microbial biomass in savannas of Central Brazil

    Directory of Open Access Journals (Sweden)

    Nardoto Gabriela Bielefeld

    2003-01-01

    Full Text Available The objective of this work was to study the effects of fire on net N mineralization and soil microbial biomass in burned and unburned cerrado stricto sensu sites. The study was carried out from April 1998 to April 2000. The pH values were significantly higher in the burned site while soil moisture content was significantly higher in the unburned site (P<0.05. The soil C/N ratio was 22/1 and the available NO3-N ranged between 1.5 and 2.8 mg kg-¹ dry weight. However, the NH4-N concentration ranged between 3 and 34 mg kg-1 dry weight in the burned site and between 3 and 22 mg kg-1 dry weight in the unburned site. The NH4-N increased after fire, but no significant changes were observed for NO3-N (P<0.05. The NO3-N accumulation occurred in short periods during the rainy season. The rates of net N mineralization increased during the rainy season while reductions in soil microbial biomass were observed at both sites. This suggested that the peak in microbial activities occurred with the first rain events, with an initial net immobilization followed by net mineralization. Both sites presented the same pattern for mineralization/immobilization, however, the amount of inorganic-N cycled annually in unburned site was 14.7 kg ha-1 per year while the burned site presented only 3.8 kg ha-¹ of inorganic-N, one year after the burning.

  6. Long-term multifactorial climate change impacts on mesofaunal biomass and nitrogen content

    DEFF Research Database (Denmark)

    Madsen, Mette Vestergård; Dyrnum, Kristine; Michelsen, Anders

    2015-01-01

    increased at elevated CO2, or tended do so. In contrast, enchytraeid N content decreased at elevated CO2. Soil microbial biomass N pool and litter C:N ratio also increased with elevated CO2, which suggests that mite biomasses are more coupled to microbial biomass, whereas enchytraeid biomass to a larger...... extent is governed by litter nitrogen concentration, i.e. litter quality. Structural equation modelling confirmed the positive coupling between soil microbial N content and oribatid biomass and further between oribatid and mesostigmatic biomass. The SEM also revealed a negative relationship between...... microbial N content and enchytraeid biomass. The biomass of all mesofaunal groups was reduced by spring drought, especially when combined with warming. Enchytraeid and especially collembolan biomass suffered greater drought declines than mite biomasses. We conclude that under long-term elevated CO2 exposure...

  7. Live Cell Discovery of Microbial Vitamin Transport and Enzyme-Cofactor Interactions

    Energy Technology Data Exchange (ETDEWEB)

    Anderson, Lindsey N.; Koech, Phillip K.; Plymale, Andrew E.; Landorf, Elizabeth V.; Konopka, Allan; Collart, Frank; Lipton, Mary S.; Romine, Margaret F.; Wright, Aaron T.

    2016-02-02

    The rapid completion of microbial genomes is inducing a conundrum in functional gene discovery. Novel methods are critically needed to shorten the gap between characterizing a microbial genome and experimentally validating bioinformatically-predicted functions. Of particular importance are transport mechanisms, used to shuttle nutrients and metabolites across cell mem-branes, such as B vitamins, which are indispensable to metabolic reactions crucial to the survival of diverse microbes ranging from members of environmental microbial communities to human pathogens. Methods to accurately assign function and specificity for a wide range of experimentally unidentified and/or predicted membrane-embedded transport proteins, and characterization of intra-cellular enzyme-cofactor/nutrient associations are needed to enable a significantly improved understanding of microbial biochemis-try and physiology, how microbes associate with others, and how they sense and respond to environmental perturbations. Chemical probes derived from B vitamins B1, B2, and B7 have allowed us to experimentally address the aforementioned needs by identifying B vitamin transporters and intracellular protein-cofactor associations through live cell labeling of the filamentous anoxygenic pho-toheterotroph, Chloroflexus aurantiacus J-10-fl, known for both B vitamin biosynthesis and environmental salvage. Our probes provide a unique opportunity to directly link cellular activity and protein function back to ecosystem and/or host dynamics by iden-tifying B vitamin transport and disposition mechanisms required for survival.

  8. Reduced Nutrient Excretion and Environmental Microbial Load with the Addition of a Combination of Enzymes and Direct-Fed Microbials to the Diet of Broiler Chickens

    Directory of Open Access Journals (Sweden)

    MFFM Praes

    2016-03-01

    Full Text Available Abstract This study evaluated the effects of the dietary inclusion of an enzyme blend and a direct-fed microbials in broiler diets on litter production and quality. In total, 900 Cobb 500(r broiler chicks were distributed according to a completely randomized design into 4 treatments and 9 replicates of 25 birds each. Broilers were reared from 1 to 42 days of age. The treatments consisted of the following diets: NC: negative control; DFM: NC + 500 ppm of direct-fed microbials product (DFM, containing Bacillus subtilis and Bacillus licheniformis; ENZ: diet formulated with an enzyme blend (20 ppm phytase, 200 ppm protease and 200 ppm of xylanase; DFM+E: ENZ + DFM. Birds and litter were weighed at the start and end of the rearing period, for litter production and waste ratio (Rw determination. Litter samples were analyzed for dry matter (DM content, total and thermotolerant coliform counts, nutrient composition (nitrogen (N, phosphorous (P and potassium (K, and fiber fraction (neutral detergent fiber (NDF, acid detergent fiber (ADF and lignin. The dietary inclusion of the evaluated additivesdid not influence litter production or Rw; however, ADF (%, NDF (kg and kg/kg DM litter, and total and thermotolerant coliform counts were reduced, and N content increased in the litter. The diets containing enzymes (ENZ and DFM+E reduced litter P content. The addition of exogenous enzymes and their combination with a DFM based on Bacillus spp .Did not affect waste production, and reduced litter microbial load, and the contents of P and insoluble fiber in the litter.

  9. Thalassic biogas production from sea wrack biomass using different microbial seeds: cow manure, marine sediment and sea wrack-associated microflora.

    Science.gov (United States)

    Marquez, Gian Powell B; Reichardt, Wolfgang T; Azanza, Rhodora V; Klocke, Michael; Montaño, Marco Nemesio E

    2013-04-01

    Sea wrack (dislodged sea grasses and seaweeds) was used in biogas production. Fresh water scarcity in island communities where sea wrack could accumulate led to seawater utilization as liquid substrate. Three microbial seeds cow manure (CM), marine sediment (MS), and sea wrack-associated microflora (SWA) were explored for biogas production. The average biogas produced were 2172±156 mL (MS), 1223±308 mL (SWA) and 551±126 mL (CM). Though methane potential (396.9 mL(CH4) g(-1) volatile solid) computed from sea wrack proximate values was comparable to other feedstocks, highest methane yield was low (MS=94.33 mL(CH4) g(-1) VS). Among the microbial seeds, MS proved the best microbial source in utilizing sea wrack biomass and seawater. However, salinity (MS=42‰) observed exceeded average seawater salinity (34‰). Hence, methanogenic activity could have been inhibited. This is the first report on sea wrack biomass utilization for thalassic biogas production. Copyright © 2013 Elsevier Ltd. All rights reserved.

  10. Effects of phosphorus addition on soil microbial biomass and community composition in three forest types in tropical China

    DEFF Research Database (Denmark)

    Liu, Lei; Gundersen, Per; Zhang, Tao

    2012-01-01

    Elevated nitrogen (N) deposition in humid tropical regions may aggravate phosphorus (P) deficiency in forest on old weathered soil found in these regions. From January 2007 to August 2009, we studied the responses of soil microbial biomass and community composition to P addition (in two monthly...

  11. Fungal Biomass Protein Production from Trichoderma harzianum Using Rice Polishing.

    Science.gov (United States)

    Ahmed, Sibtain; Mustafa, Ghulam; Arshad, Muhammad; Rajoka, Muhammad Ibrahim

    2017-01-01

    Industrially important enzymes and microbial biomass proteins have been produced from fungi for more than 50 years. High levels of crude protein as much as 45% are present in fungal biomass with balanced essential amino acids. The aim of this study was to access the potential of Trichoderma harzianum to produce fungal biomass protein from rice polishings. Maximum biomass yield was obtained at 5% (w/v) rice polishings after 72 h of incubation at 28°C at pH 4. Carbon and nitrogen ratio of 20 : 1 gave significantly higher production of fungal biomass protein. The FBP in the 75 L fermenter contained 49.50% crude protein, 32.00% true protein, 19.45% crude fiber, 9.62% ash, 11.5% cellulose content, and 0.325% RNA content. The profile of amino acids of final FBP exhibited that all essential amino acids were present in great quantities. The FBP produced by this fungus has been shown to be of good nutritional value for supplementation to poultry. The results presented in this study have practical implications in that the fungus T. harzianum could be used successfully to produce fungal biomass protein using rice polishings.

  12. Effects of nitrogen and phosphorus additions on soil microbial biomass and community structure in two reforested tropical forests.

    Science.gov (United States)

    Liu, Lei; Gundersen, Per; Zhang, Wei; Zhang, Tao; Chen, Hao; Mo, Jiangming

    2015-09-23

    Elevated nitrogen (N) deposition may aggravate phosphorus (P) deficiency in forests in the warm humid regions of China. To our knowledge, the interactive effects of long-term N deposition and P availability on soil microorganisms in tropical replanted forests remain unclear. We conducted an N and P manipulation experiment with four treatments: control, N addition (15 g N m(-2)·yr(-1)), P addition (15 g P m(-2)·yr(-1)), and N and P addition (15 + 15 g N and P m(-2)·yr(-1), respectively) in disturbed (planted pine forest with recent harvests of understory vegetation and litter) and rehabilitated (planted with pine, but mixed with broadleaf returning by natural succession) forests in southern China. Nitrogen addition did not significantly affect soil microbial biomass, but significantly decreased the abundance of gram-negative bacteria PLFAs in both forest types. Microbial biomass increased significantly after P addition in the disturbed forest but not in the rehabilitated forest. No interactions between N and P additions on soil microorganisms were observed in either forest type. Our results suggest that microbial growth in replanted forests of southern China may be limited by P rather than by N, and this P limitation may be greater in disturbed forests.

  13. D:L-AMINO Acids and the Turnover of Microbial Biomass

    Science.gov (United States)

    Lomstein, B. A.; Braun, S.; Mhatre, S. S.; Jørgensen, B. B.

    2015-12-01

    Decades of ocean drilling have demonstrated wide spread microbial life in deep sub-seafloor sediment, and surprisingly high microbial cell numbers. Despite the ubiquity of life in the deep biosphere, the large community sizes and the low energy fluxes in the vast buried ecosystem are still poorly understood. It is not know whether organisms of the deep biosphere are specifically adapted to extremely low energy fluxes or whether most of the observed cells are in a maintenance state. Recently we developed and applied a new culture independent approach - the D:L-amino acid model - to quantify the turnover times of living microbial biomass, microbial necromass and mean metabolic rates. This approach is based on the built-in molecular clock in amino acids that very slowly undergo chemical racemization until they reach an even mixture of L- and D- forms, unless microorganisms spend energy to keep them in the L-form that dominates in living organisms. The approach combines sensitive analyses of amino acids, the unique bacterial endospore marker (dipicolinic acid) with racemization dynamics of stereo-isomeric amino acids. Based on a heating experiment, we recently reported kinetic parameters for racemization of aspartic acid, glutamic acid, serine and alanine in bulk sediment from Aarhus Bay, Denmark. The obtained racemization rate constants were faster than the racemization rate constants of free amino acids, which we have previously applied in Holocene sediment from Aarhus Bay and in up to 10 mio yr old sediment from ODP Leg 201. Another important input parameter for the D:L-amino acid model is the cellular carbon content. It has recently been suggested that the cellular carbon content most likely is lower than previously thought. In recognition of these new findings, previously published data based on the D:L-amino acid model were recalculated and will be presented together with new data from an Arctic Holocene setting with constant sub-zero temperatures.

  14. Impact of heavy metal on activity of some microbial enzymes in the riverbed sediments: Ecotoxicological implications in the Ganga River (India).

    Science.gov (United States)

    Jaiswal, Deepa; Pandey, Jitendra

    2018-04-15

    We studied the extracellular enzyme activity (EEA) in the riverbed sediment along a 518km gradient of the Ganga River receiving carbon and nutrient load from varied human sources. Also, we tested, together with substrate-driven stimulation, if the heavy metal accumulated in the sediment inhibits enzyme activities. Because pristine values are not available, we considered Dev Prayag, a least polluted site located 624km upstream to main study stretch, as a reference site. There were distinct increases in enzyme activities in the sediment along the study gradient from Dev Prayag, however, between-site differences were in concordance with sediment carbon(C), nitrogen (N) and phosphorus (P). Fluorescein diacetate hydrolysis (FDAase), β-glucosidase (Glu) and protease activities showed positive correlation with C, N and P while alkaline phosphatase was found negatively correlated with P. Enzyme activities were found negatively correlated with heavy metal, although ecological risk index (E R i ) varied with site and metal species. Dynamic fit curves showed significant positive correlation between heavy metal and microbial metabolic quotient (qCO 2 ) indicating a decrease in microbial activity in response to increasing heavy metal concentrations. This study forms the first report linking microbial enzyme activities to regional scale sediment heavy metal accumulation in the Ganga River, suggests that the microbial enzyme activities in the riverbed sediment were well associated with the proportion of C, N and P and appeared to be a sensitive indicator of C, N and P accumulation in the river. Heavy metal accumulated in the sediment inhibits enzyme activities, although C rich sediment showed relatively low toxicity due probably to reduced bioavailability of the metal. The study has relevance from ecotoxicological as well as from biomonitoring perspectives. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. Biosynthesis of the enzymes of the cellulase system by T. Reesei QM 9414 in the presence of sophorose

    Science.gov (United States)

    Gritzali, M.

    1982-12-01

    As conventional, nonrenewable energy sources are rapidly depleted and it was necessary to search for alternative sources of energy. It was increasingly apparent that biomass and waste are alternatives well worth exploring. The sources of biomass and wastes that considered for conversion to useful products are quite diverse, but the most abundant constituent of almost every type is cellulose. Cellulose is cleanly converted to soluble fermentable sugars enzymatically, and cellulose enzymes were isolated from a number of microbial sources. It is generally agreed that the most effective system of enzymes for the conversion of cellulose to glucose is produced by species of the imperfect fungus Trichoderma. The mutant organism Trichoderma reesei QM 9414 is among the best producers of high levels of enzymes; these are extracellular and have carbonhydrate covalently bound to the peptide. Trichoderma produces three types of enzymes which, in a sequential and cooperative manner, convert cellulose to soluble oligosaccharides and glucose.

  16. Biomass recalcitrance

    DEFF Research Database (Denmark)

    Felby, Claus

    2009-01-01

    Alternative and renewable fuels derived from lignocellulosic biomass offer a promising alternative to conventional energy sources, and provide energy security, economic growth, and environmental benefits. However, plant cell walls naturally resist decomposition from microbes and enzymes - this co......Alternative and renewable fuels derived from lignocellulosic biomass offer a promising alternative to conventional energy sources, and provide energy security, economic growth, and environmental benefits. However, plant cell walls naturally resist decomposition from microbes and enzymes...... - this collective resistance is known as "biomass recalcitrance." Breakthrough technologies are needed to overcome barriers to developing cost-effective processes for converting biomass to fuels and chemicals. This book examines the connection between biomass structure, ultrastructure, and composition......, to resistance to enzymatic deconstruction, with the aim of discovering new cost-effective technologies for biorefineries. It contains chapters on topics extending from the highest levels of biorefinery design and biomass life-cycle analysis, to detailed aspects of plant cell wall structure, chemical treatments...

  17. Influence of heavy metals and PCBs pollution on the enzyme activity and microbial community of paddy soils around an e-waste recycling workshop.

    Science.gov (United States)

    Tang, Xianjin; Hashmi, Muhammad Z; Long, Dongyan; Chen, Litao; Khan, Muhammad I; Shen, Chaofeng

    2014-03-14

    Due to the emerging environmental issues related to e-waste there is concern about the quality of paddy soils near e-waste workshops. The levels of heavy metals and PCBs and their influence on the enzyme activity and microbial community of paddy soils obtained from the immediate vicinity of an e-waste workshop were investigated in the present study. The results indicated that the heavy metal and PCB pollution did not differ significantly with an increase of the sampling point distances (5 to 30 m). The concentration of Cd (2.16 mg·kg-1) and Cu (69.2 mg·kg-1) were higher, and the PCB pollution was also serious, ranging from 4.9 to 21.6 μg·kg-1. The highest enzyme activity was found for urease compared to phosphatase and catalase, and a fluctuating trend in soil enzyme activity was observed in soils from different sampling sites. The microbial analysis revealed that there was no apparent correlation between the microbial community and the pollutants. However, a slight influence for soil microbial communities could be found based on DGGE, the Shannon index and PCA analysis. The present study suggests that the contamination stress of heavy metals and PCBs might have a slight influence on microbial activity in paddy soils. This study provides the baseline data for enzyme activities and microbial communities in paddy soil under the influence of mixed contamination.

  18. Influence of Heavy Metals and PCBs Pollution on the Enzyme Activity and Microbial Community of Paddy Soils around an E-Waste Recycling Workshop

    Directory of Open Access Journals (Sweden)

    Xianjin Tang

    2014-03-01

    Full Text Available Due to the emerging environmental issues related to e-waste there is concern about the quality of paddy soils near e-waste workshops. The levels of heavy metals and PCBs and their influence on the enzyme activity and microbial community of paddy soils obtained from the immediate vicinity of an e-waste workshop were investigated in the present study. The results indicated that the heavy metal and PCB pollution did not differ significantly with an increase of the sampling point distances (5 to 30 m. The concentration of Cd (2.16 mg·kg−1 and Cu (69.2 mg·kg−1 were higher, and the PCB pollution was also serious, ranging from 4.9 to 21.6 μg·kg−1. The highest enzyme activity was found for urease compared to phosphatase and catalase, and a fluctuating trend in soil enzyme activity was observed in soils from different sampling sites. The microbial analysis revealed that there was no apparent correlation between the microbial community and the pollutants. However, a slight influence for soil microbial communities could be found based on DGGE, the Shannon index and PCA analysis. The present study suggests that the contamination stress of heavy metals and PCBs might have a slight influence on microbial activity in paddy soils. This study provides the baseline data for enzyme activities and microbial communities in paddy soil under the influence of mixed contamination.

  19. Influence of Heavy Metals and PCBs Pollution on the Enzyme Activity and Microbial Community of Paddy Soils around an E-Waste Recycling Workshop

    Science.gov (United States)

    Tang, Xianjin; Hashmi, Muhammad Z.; Long, Dongyan; Chen, Litao; Khan, Muhammad I.; Shen, Chaofeng

    2014-01-01

    Due to the emerging environmental issues related to e-waste there is concern about the quality of paddy soils near e-waste workshops. The levels of heavy metals and PCBs and their influence on the enzyme activity and microbial community of paddy soils obtained from the immediate vicinity of an e-waste workshop were investigated in the present study. The results indicated that the heavy metal and PCB pollution did not differ significantly with an increase of the sampling point distances (5 to 30 m). The concentration of Cd (2.16 mg·kg−1) and Cu (69.2 mg·kg−1) were higher, and the PCB pollution was also serious, ranging from 4.9 to 21.6 μg·kg−1. The highest enzyme activity was found for urease compared to phosphatase and catalase, and a fluctuating trend in soil enzyme activity was observed in soils from different sampling sites. The microbial analysis revealed that there was no apparent correlation between the microbial community and the pollutants. However, a slight influence for soil microbial communities could be found based on DGGE, the Shannon index and PCA analysis. The present study suggests that the contamination stress of heavy metals and PCBs might have a slight influence on microbial activity in paddy soils. This study provides the baseline data for enzyme activities and microbial communities in paddy soil under the influence of mixed contamination. PMID:24637907

  20. Earthworms facilitate the stabilization of pelletized dewatered sludge through shaping microbial biomass and activity and community.

    Science.gov (United States)

    Fu, Xiaoyong; Cui, Guangyu; Huang, Kui; Chen, Xuemin; Li, Fusheng; Zhang, Xiaoyu; Li, Fei

    2016-03-01

    In this study, the effect of earthworms on microbial features during vermicomposting of pelletized dewatered sludge (PDS) was investigated through comparing two degradation systems with and without earthworm E isenia fetida involvement. After 60 days of experimentation, a relatively stable product with low organic matter and high nitrate and phosphorous was harvested when the earthworms were involved. During the process, earthworms could enhance microbial activity and biomass at the initial stage and thus accelerating the rapid decomposition of PDS. The end products of vermicomposting allowed the lower values of bacterial and eukaryotic densities comparison with those of no earthworm addition. In addition, the presence of earthworms modified the bacterial and fungal diversity, making the disappearances of some pathogens and specific decomposing bacteria of recalcitrant substrates in the vermicomposting process. This study evidences that earthworms can facilitate the stabilization of PDS through modifying microbial activity and number and community during vermicomposting.

  1. Construction of efficient xylose utilizing Pichia pastoris for industrial enzyme production.

    Science.gov (United States)

    Li, Pengfei; Sun, Hongbing; Chen, Zao; Li, Yin; Zhu, Taicheng

    2015-02-21

    Cellulosic biomass especially agricultural/wood residues can be utilized as feedstock to cost-effectively produce fuels, chemicals and bulk industrial enzymes, which demands xylose utilization from microbial cell factories. While previous works have made significant progress in improving microbial conversion of xylose into fuels and chemicals, no study has reported the engineering of efficient xylose utilizing protein expression systems for the purpose of producing industrial enzymes. In this work, using Pichia pastoris as an example, we demonstrated the successful engineering of xylose metabolizing ability into of protein expression systems. A heterologous XI (xylose isomerase) pathway was introduced into P. pastoris GS115 by overexpressing the Orpinomyces spp. XI or/and the endogenous XK (xylulokinase) gene, and evolutionary engineering strategies were also applied. Results showed that the XI pathway could be functionally expressed in P. pastoris. After 50 generation of sequential batch cultivation, a set of domesticated recombinant P. pastoris strains with different performance metrics on xylose were obtained. One evolved strain showed the highest xylose assimilation ability, whose cell yield on xylose can even be comparable to that on glucose or glycerol. This strain also showed significantly increased β-mannanase production when cultured on xylose medium. Furthermore, transcription analysis of xylose pathway genes suggested that overexpression of XI and XK might be the key factors affecting effective xylose assimilation. To our best knowledge, this study is the first work demonstrating the construction of efficient xylose utilizing P. pastoris strains, thus providing a basis for using cellulosic biomass for bulk industrial enzyme production.

  2. Effects of lead contamination on soil enzymatic activities, microbial biomass, and rice physiological indices in soil-lead-rice (Oryza sativa L.) system.

    Science.gov (United States)

    Zeng, Lu S; Liao, Min; Chen, Cheng L; Huang, Chang Y

    2007-05-01

    The effect of lead (Pb) treatment on the soil enzymatic activities, soil microbial biomass, rice physiological indices and rice biomass were studied in a greenhouse pot experiment. Six levels of Pb viz. 0(CK), 100, 300, 500, 700, 900 mg/kg soil were applied in two types of paddy soils. The results showed that Pb treatment had a stimulating effect on soil enzymatic activities and microbial biomass carbon (Cmic) at low concentration and an inhibitory influence at higher concentration. The degree of influence on enzymatic activities and Cmic by Pb was related to the clay and organic matter contents of the soils. When the Pb treatment was raised to the level of 500 mg/kg, ecological risk appeared both to soil microorganisms and plants. The results also revealed a consistent trend of increased chlorophyll contents and rice biomass initially, maximum at a certain Pb treatment, and then decreased gradually with the increase in Pb concentration. Pb was effective in inducing proline accumulation and its toxicity causes oxidative stress in rice plants. Therefore, it was concluded that soil enzymatic activities, Cmic and rice physiological indices, could be sensitive indicators to reflect environmental stress in soil-lead-rice system.

  3. The use of N 15 for studying the relation between the development of the microbial biomass and the remineralization of nitrogen fertilizer by rice

    Energy Technology Data Exchange (ETDEWEB)

    Al-Chater, M S [King Faisal University - Faculty of Agriculture and Food Science Department of Soil and Water Al-Hassa, (Saudi Arabia)

    1995-10-01

    The study aims at determining the mechanism of secondary mineralization of fertilizer N in the form of (N{sup 15} H{sub 14})2 SO{sub 4} which was transformed to the organic N using soil samples taken after a period of 16 weeks from the series of pot experiments. The development of the microbial biomass was studied using the method of Jenkinson and Powlson(1976) which was modified by Bottner et al (1984). Nitrogen mineralization was determined using the Stanford and Smith (1972) method. The study indicated that the values of mineralization constant varied from 0.0012 to 0.027 in different treatments. Accordingly, the half-life of microbial biomass ranged from 6 to 7 months, indicating fast transformation of microbial N as compared with total organic N in the soil which ranged between 7 to 12 years.1 fig., 3 tabs.

  4. Microbial respiration and extracellular enzyme activity in sediments from the Gulf of Mexico hypoxic zone

    Science.gov (United States)

    This study explores the relationship between sediment chemistry (TC, TN, TP) and microbial respiration (DHA) and extracellular enzyme activity (EEA) across the Gulf of Mexico (GOM) hypoxic zone. TC, TN, and TP were all positively correlated with each other (r=0.19-0.68). DHA was ...

  5. Optimization studies for the bioconversion of Jerusalem artichoke tubers to ethanol and microbial biomass

    Energy Technology Data Exchange (ETDEWEB)

    Margaritis, A.; Bajpai, P.; Cannell, E.

    1981-01-01

    A total of 8 yeast and other microbial cultures were grown in the extract derived from the tubers of Jerusalem artichoke (Helianthus tuberosus) and screened according to the following optimization criteria: rates and yields of ethanol production, rates and yields of biomass production, and percent of original sugars utilized during fermentation. Batch growth kinetic parameters were also determined for the cultures studied. Kluyveromyces marxianus UCD (FST) 55-82 had the highest specific growth rate, 0.41/h, with a high ethanol yield, 88% of theoretical.

  6. Influence of mechanical disintegration on the microbial growth of aerobic sludge biomass: A comparative study of ultrasonic and shear gap homogenizers by oxygen uptake measurements.

    Science.gov (United States)

    Divyalakshmi, P; Murugan, D; Sivarajan, M; Saravanan, P; Lajapathi Rai, C

    2015-11-01

    Wastewater treatment plant incorporates physical, chemical and biological processes to treat and remove the contaminants. The main drawback of conventional activated sludge process is the huge production of excess sludge, which is an unavoidable byproduct. The treatment and disposal of excess sludge costs about 60% of the total operating cost. The ideal way to reduce excess sludge production during wastewater treatment is by preventing biomass formation within the aerobic treatment train rather than post treatment of the generated sludge. In the present investigation two different mechanical devices namely, Ultrasonic and Shear Gap homogenizers have been employed to disintegrate the aerobic biomass. This study is intended to restrict the multiplication of microbial biomass and at the same time degrade the organics present in wastewater by increasing the oxidative capacity of microorganisms. The disintegrability on biomass was determined by biochemical methods. Degree of inactivation provides the information on inability of microorganisms to consume oxygen upon disruption. The soluble COD quantifies the extent of release of intra cellular compounds. The participation of disintegrated microorganism in wastewater treatment process was carried out in two identical respirometeric reactors. The results show that Ultrasonic homogenizer is very effective in the disruption of microorganisms leading to a maximum microbial growth reduction of 27%. On the other hand, Shear gap homogenizer does not favor the sludge growth reduction rather it facilitates the growth. This study also shows that for better microbial growth reduction, floc size reduction alone is not sufficient but also microbial disruption is essential. Copyright © 2015 Elsevier Inc. All rights reserved.

  7. Characterisation of microbial biocoenosis in vertical subsurface flow constructed wetlands

    International Nuclear Information System (INIS)

    Tietz, Alexandra; Kirschner, Alexander; Langergraber, Guenter; Sleytr, Kirsten; Haberl, Raimund

    2007-01-01

    In this study a quantitative description of the microbial biocoenosis in subsurface vertical flow constructed wetlands fed with municipal wastewater was carried out. Three different methods (substrate induced respiration, ATP measurement and fumigation-extraction) were applied to measure the microbial biomass at different depths of planted and unplanted systems. Additionally, bacterial biomass was determined by epifluorescence microscopy and productivity was measured via 14 C leucine incorporation into bacterial biomass. All methods showed that > 50% of microbial biomass and bacterial activity could be found in the first cm and about 95% in the first 10 cm of the filter layer. Bacterial biomass in the first 10 cm of the filter body accounted only for 16-19% of the total microbial biomass. Whether fungi or methodical uncertainties are mainly responsible for the difference between microbial and bacterial biomass remains to be examined. A comparison between the purification performance of planted and unplanted pilot-scale subsurface vertical flow constructed wetlands (PSCWs) showed no significant difference with the exception of the reduction of enterococci. The microbial biomass in all depths of the filter body was also not different in planted and unplanted systems. Compared with data from soils the microbial biomass in the PSCWs was high, although the specific surface area of the used sandy filter material available for biofilm growth was lower, especially in the beginning of the set-up of the PSCWs, due to missing clay and silt fraction

  8. Influence of altered precipitation pattern on greenhouse gas emissions and soil enzyme activities in Pannonian soils

    Science.gov (United States)

    Forstner, Stefan Johannes; Michel, Kerstin; Berthold, Helene; Baumgarten, Andreas; Wanek, Wolfgang; Zechmeister-Boltenstern, Sophie; Kitzler, Barbara

    2013-04-01

    Precipitation patterns are likely to be altered due to climate change. Recent models predict a reduction of mean precipitation during summer accompanied by a change in short-term precipitation variability for central Europe. Correspondingly, the risk for summer drought is likely to increase. This may especially be valid for regions which already have the potential for rare, but strong precipitation events like eastern Austria. Given that these projections hold true, soils in this area will receive water irregularly in few, heavy rainfall events and be subjected to long-lasting dry periods in between. This pattern of drying/rewetting can alter soil greenhouse gas fluxes, creating a potential feedback mechanism for climate change. Microorganisms are the key players in most soil carbon (C) and nitrogen (N) transformation processes including greenhouse gas exchange. A conceptual model proposed by Schimel and colleagues (2007) links microbial stress-response physiology to ecosystem-scale biogeochemical processes: In order to cope with decreasing soil water potential, microbes modify resource allocation patterns from growth to survival. However, it remains unclear how microbial resource acquisition via extracellular enzymes and microbial-controlled greenhouse gas fluxes respond to water stress induced by soil drying/rewetting. We designed a laboratory experiment to test for effects of multiple drying/rewetting cycles on soil greenhouse gas fluxes (CO2, CH4, N2O, NO), microbial biomass and extracellular enzyme activity. Three soils representing the main soil types of eastern Austria were collected in June 2012 at the Lysimeter Research Station of the Austrian Agency for Health and Food Safety (AGES) in Vienna. Soils were sieved to 2mm, filled in steel cylinders and equilibrated for one week at 50% water holding capacity (WHC) for each soil. Then soils were separated into two groups: One group received water several times per week (C=control), the other group received

  9. Decomposition of organic matter labelled with 14C, and microbial biomass formation in an acid soil from Piracicaba, Sao Paulo State, Brazil

    International Nuclear Information System (INIS)

    Cerri, C.C.; Volkoff, B.

    1982-01-01

    An experiment was carried out under laboratory conditions (25 0 C, 50% field capacity), using 14 C labelled sugar cane leaves, which were incorporated to a medium texture Dark-Red Latosol. The equivalent of 1 mg C/g was added to soil containing 14 mg native soil carbon. CO 2 release and microbial biomass were periodically determined. After 67 days incubation it was noted that release of native carbon was greater from soil with plant material than the control (724 μg C/g vs 424 μg/g) and that 25% of the incorporated carbon had been mineralized. Addition of plant material doubled the soil microbial biomass, which was formed due to plant material addition rather than to native soil carbon. (Author) [pt

  10. Relationships between waste physicochemical properties, microbial activity and vegetation at coal ash and sludge disposal sites.

    Science.gov (United States)

    Woch, Marcin W; Radwańska, Magdalena; Stanek, Małgorzata; Łopata, Barbara; Stefanowicz, Anna M

    2018-06-11

    The aim of the study was to assess the relationships between vegetation, physicochemical and microbial properties of substrate at coal ash and sludge disposal sites. The study was performed on 32 plots classified into 7 categories: dried ash sedimentation ponds, dominated by a grass Calamagrostis epigejos (AH-Ce), with the admixture of Pinus sylvestris (AH-CePs) or Robinia pseudoacacia (AH-CeRp), dry ash landfill dominated by Betula pendula and Pinus sylvestris (AD-BpPs) or Salix viminalis (AD-Sv) and coal sludge pond with drier parts dominated by Tussilago farfara (CS-Tf) and the wetter ones by Cyperus flavescens (CS-Cf). Ash sites were covered with soil layer imported as a part of technical reclamation. Ash had relatively high concentrations of some alkali and alkaline earth metals, Mn and pH, while coal sludge had high water and C, S, P and K contents. Concentrations of heavy metals were lower than allowable limits in all substrate types. Microbial biomass and, particularly, enzymatic activity in ash and sludge were generally low. The only exception were CS-Tf plots characterized by the highest microbial biomass, presumably due to large deposits of organic matter that became available for aerobic microbial biomass when water level fell. The properties of ash and sludge adversely affected microbial biomass and enzymatic activity as indicated by significant negative correlations between the content of alkali/alkaline earth metals, heavy metals, and macronutrients with enzymatic activity and/or microbial biomass, as well as positive correlations of these parameters with metabolic quotient (qCO 2 ). Plant species richness and cover were relatively high, which may be partly associated with alleviating influence of soil covering the ash. The effect of the admixture of R. pseudoacacia or P. sylvestris to stands dominated by C. epigejos was smaller than expected. The former species increased NNH 4 , NNO 3 and arylsulfatase activity, while the latter reduced activity of

  11. Does microbial biomass affect pelagic ecosystem efficiency? An experimental study.

    Science.gov (United States)

    Wehr, J D; Le, J; Campbell, L

    1994-01-01

    Bacteria and other microorganisms in the pelagic zone participate in the recycling of organic matter and nutrients within the water column. The microbial loop is thought to enhance ecosystem efficiency through rapid recycling and reduced sinking rates, thus reducing the loss of nutrients contained in organisms remaining within the photic zone. We conducted experiments with lake communities in 5400-liter mesocosms, and measured the flux of materials and nutrients out of the water column. A factorial design manipulated 8 nutrient treatments: 4 phosphorus levels × 2 nitrogen levels. Total sedimentation rates were greatest in high-N mesocosms; within N-surplus communities, [Symbol: see text]1 µM P resulted in 50% increase in total particulate losses. P additions without added N had small effects on nutrient losses from the photic zone; +2 µM P tanks received 334 mg P per tank, yet after 14 days lost only 69 mg more particulate-P than did control communities. Nutrient treatments resulted in marked differences in phytoplankton biomass (twofold N effect, fivefold P effect in +N mesocosms only), bacterioplankton densities (twofold N-effect, twofold P effects in -N and +N mesocosms), and the relative importance of autotrophic picoplankton (maximum in high NY mesocosms). Multiple regression analysis found that of 8 plankton and water chemistry variables, the ratio of autotrophic picoplankton to total phytoplankton (measured as chlorophyll α) explained the largest portion of the total variation in sedimentation loss rates (65% of P-flux, 57% of N-flux, 26% of total flux). In each case, systems with greater relative importance of autotrophic picoplankton had significantly reduced loss rates. In contrast, greater numbers of planktonic bacteria were associated with increased sedimentation rates and lower system efficiency. We suggest that different microbial components may have contrasting effects on the presumed enhanced efficiency provided by the microbial loop.

  12. An Integrated Metagenomics/Metaproteomics Investigation of the Microbial Communities and Enzymes in Solid-state Fermentation of Pu-erh tea

    Science.gov (United States)

    Zhao, Ming; Zhang, Dong-lian; Su, Xiao-qin; Duan, Shuang-mei; Wan, Jin-qiong; Yuan, Wen-xia; Liu, Ben-ying; Ma, Yan; Pan, Ying-hong

    2015-01-01

    Microbial enzymes during solid-state fermentation (SSF), which play important roles in the food, chemical, pharmaceutical and environmental fields, remain relatively unknown. In this work, the microbial communities and enzymes in SSF of Pu-erh tea, a well-known traditional Chinese tea, were investigated by integrated metagenomics/metaproteomics approach. The dominant bacteria and fungi were identified as Proteobacteria (48.42%) and Aspergillus (94.98%), through pyrosequencing-based analyses of the bacterial 16S and fungal 18S rRNA genes, respectively. In total, 335 proteins with at least two unique peptides were identified and classified into 28 Biological Processes and 35 Molecular Function categories using a metaproteomics analysis. The integration of metagenomics and metaproteomics data demonstrated that Aspergillus was dominant fungus and major host of identified proteins (50.45%). Enzymes involved in the degradation of the plant cell wall were identified and associated with the soft-rotting of tea leaves. Peroxiredoxins, catalase and peroxidases were associated with the oxidation of catechins. In conclusion, this work greatly advances our understanding of the SSF of Pu-erh tea and provides a powerful tool for studying SSF mechanisms, especially in relation to the microbial communities present. PMID:25974221

  13. The Role of Microbial Community Composition in Controlling Soil Respiration Responses to Temperature.

    Science.gov (United States)

    Auffret, Marc D; Karhu, Kristiina; Khachane, Amit; Dungait, Jennifer A J; Fraser, Fiona; Hopkins, David W; Wookey, Philip A; Singh, Brajesh K; Freitag, Thomas E; Hartley, Iain P; Prosser, James I

    2016-01-01

    Rising global temperatures may increase the rates of soil organic matter decomposition by heterotrophic microorganisms, potentially accelerating climate change further by releasing additional carbon dioxide (CO2) to the atmosphere. However, the possibility that microbial community responses to prolonged warming may modify the temperature sensitivity of soil respiration creates large uncertainty in the strength of this positive feedback. Both compensatory responses (decreasing temperature sensitivity of soil respiration in the long-term) and enhancing responses (increasing temperature sensitivity) have been reported, but the mechanisms underlying these responses are poorly understood. In this study, microbial biomass, community structure and the activities of dehydrogenase and β-glucosidase enzymes were determined for 18 soils that had previously demonstrated either no response or varying magnitude of enhancing or compensatory responses of temperature sensitivity of heterotrophic microbial respiration to prolonged cooling. The soil cooling approach, in contrast to warming experiments, discriminates between microbial community responses and the consequences of substrate depletion, by minimising changes in substrate availability. The initial microbial community composition, determined by molecular analysis of soils showing contrasting respiration responses to cooling, provided evidence that the magnitude of enhancing responses was partly related to microbial community composition. There was also evidence that higher relative abundance of saprophytic Basidiomycota may explain the compensatory response observed in one soil, but neither microbial biomass nor enzymatic capacity were significantly affected by cooling. Our findings emphasise the key importance of soil microbial community responses for feedbacks to global change, but also highlight important areas where our understanding remains limited.

  14. Cultivation of Podospora anserina on soybean hulls results in an efficient enzyme cocktail for plant biomass hydrolysis

    NARCIS (Netherlands)

    Mäkelä, Miia R; Bouzid, Ourdia; Ruiz-Robleto, J.; Post, Harm|info:eu-repo/dai/nl/341667374; Peng, Mao; Heck, Albert|info:eu-repo/dai/nl/105189332; Altelaar, Maarten|info:eu-repo/dai/nl/304833517; de Vries, Ronald P|info:eu-repo/dai/nl/186324960

    2017-01-01

    The coprophilic ascomycete fungus Podospora anserina was cultivated on three different plant biomasses, i.e. cotton seed hulls (CSH), soybean hulls (SBH) and acid-pretreated wheat straw (WS) for four days, and the potential of the produced enzyme mixtures was compared in the enzymatic

  15. Site- and horizon-specific patterns of microbial community structure and enzyme activities in permafrost-affected soils of Greenland

    Science.gov (United States)

    Gittel, Antje; Bárta, Jiří; Kohoutová, Iva; Schnecker, Jörg; Wild, Birgit; Čapek, Petr; Kaiser, Christina; Torsvik, Vigdis L.; Richter, Andreas; Schleper, Christa; Urich, Tim

    2014-01-01

    Permafrost-affected soils in the Northern latitudes store huge amounts of organic carbon (OC) that is prone to microbial degradation and subsequent release of greenhouse gasses to the atmosphere. In Greenland, the consequences of permafrost thaw have only recently been addressed, and predictions on its impact on the carbon budget are thus still highly uncertain. However, the fate of OC is not only determined by abiotic factors, but closely tied to microbial activity. We investigated eight soil profiles in northeast Greenland comprising two sites with typical tundra vegetation and one wet fen site. We assessed microbial community structure and diversity (SSU rRNA gene tag sequencing, quantification of bacteria, archaea and fungi), and measured hydrolytic and oxidative enzyme activities. Sampling site and thus abiotic factors had a significant impact on microbial community structure, diversity and activity, the wet fen site exhibiting higher potential enzyme activities and presumably being a hot spot for anaerobic degradation processes such as fermentation and methanogenesis. Lowest fungal to bacterial ratios were found in topsoils that had been relocated by cryoturbation (“buried topsoils”), resulting from a decrease in fungal abundance compared to recent (“unburied”) topsoils. Actinobacteria (in particular Intrasporangiaceae) accounted for a major fraction of the microbial community in buried topsoils, but were only of minor abundance in all other soil horizons. It was indicated that the distribution pattern of Actinobacteria and a variety of other bacterial classes was related to the activity of phenol oxidases and peroxidases supporting the hypothesis that bacteria might resume the role of fungi in oxidative enzyme production and degradation of phenolic and other complex substrates in these soils. Our study sheds light on the highly diverse, but poorly-studied communities in permafrost-affected soils in Greenland and their role in OC degradation. PMID

  16. Short-term incorporation of organic manures and biofertilizers influences biochemical and microbial characteristics of soils under an annual crop [Turmeric (Curcuma longa L.)].

    Science.gov (United States)

    Dinesh, R; Srinivasan, V; Hamza, S; Manjusha, A

    2010-06-01

    The study was conducted to determine whether short-term incorporation of organic manures and biofertilizers influence biochemical and microbial variables reflecting soil quality. For the study, soils were collected from a field experiment conducted on turmeric (Curcuma longa L.) involving organic nutrient management (ONM), chemical nutrient management (CNM) and integrated nutrient management (INM). The findings revealed that application of organic manures and biofertilizers (ONM and INM) positively influenced microbial biomass C, N mineralization, soil respiration and enzymes activities. Contrarily, greater metabolic quotient levels in CNM indicated a stressed soil microbial community. Principal component analysis indicated the strong relationship between microbial activity and the availability of labile and easily mineralizable organic matter. The findings imply that even short-term incorporation of organic manures and biofertilizers promoted soil microbial and enzyme activities and these parameters are sensitive enough to detect changes in soil quality due to short-term incorporation of biological fertilizers. (c) 2010 Elsevier Ltd. All rights reserved.

  17. Extreme CO2 disturbance and the resilience of soil microbial communities

    Science.gov (United States)

    McFarland, Jack W.; Waldrop, Mark P.; Haw, Monica

    2013-01-01

    Carbon capture and storage (CSS) technology has the potential to inadvertently release large quantities of CO2 through geologic substrates and into surrounding soils and ecosystems. Such a disturbance has the potential to not only alter the structure and function of plant and animal communities, but also soils, soil microbial communities, and the biogeochemical processes they mediate. At Mammoth Mountain, we assessed the soil microbial community response to CO2 disturbance (derived from volcanic ‘cold’ CO2) that resulted in localized tree kill; soil CO2 concentrations in our study area ranged from 0.6% to 60%. Our objectives were to examine how microbial communities and their activities are restructured by extreme CO2 disturbance, and assess the response of major microbial taxa to the reintroduction of limited plant communities following an extensive period (15–20 years) with no plants. We found that CO2-induced tree kill reduced soil carbon (C) availability along our sampling transect. In response, soil microbial biomass decreased by an order of magnitude from healthy forest to impacted areas. Soil microorganisms were most sensitive to changes in soil organic C, which explained almost 60% of the variation for microbial biomass C (MBC) along the CO2gradient. We employed phospholipid fatty acid analysis and quantitative PCR (qPCR) to determine compositional changes among microbial communities in affected areas and found substantial reductions in microbial biomass linked to the loss of soil fungi. In contrast, archaeal populations responded positively to the CO2 disturbance, presumably due to reduced competition of bacteria and fungi, and perhaps unique adaptations to energy stress. Enzyme activities important in the cycling of soil C, nitrogen (N), and phosphorus (P) declined with increasing CO2, though specific activities (per unit MBC) remained stable or increased suggesting functional redundancy among restructured communities. We conclude that both the

  18. Assessment of natural sepiolite on cadmium stabilization, microbial communities, and enzyme activities in acidic soil.

    Science.gov (United States)

    Sun, Yuebing; Sun, Guohong; Xu, Yingming; Wang, Lin; Liang, Xuefeng; Lin, Dasong; Hu, Fazhi

    2013-05-01

    A pot trial was conducted to assess the efficiency of sepiolite-induced cadmium (Cd) immobilization in ultisoils. Under Cd concentrations of 1.25, 2.5, and 5 mg kg(-1), the available Cd in the soil after the application of 1-10 % sepiolite decreased by a maximum of 44.4, 23.0, and 17.0 %, respectively, compared with no sepiolite treatments. The increase in the values of soil enzyme activities and microbial number proved that a certain metabolic recovery occurred after sepiolite treatment. The dry biomass of spinach (Spinacia oleracea) increased with increasing sepiolite concentration in the soil. However, the concentration (dry weight) of Cd in the spinach shoots decreased with the increase in sepiolite dose, with maximum reduction of 92.2, 90.0, and 84.9 %, respectively, compared with that of unamended soils. Under a Cd level of 1.25 mg kg(-1), the Cd concentration in the edible parts of spinach at 1 % sepiolite amendment was lower than 0.2 mg kg(-1) fresh weight, the maximum permissible concentration (MPC) of Cd in vegetable. Even at higher Cd concentrations (2.5 and 5 mg kg(-1)), safe spinach was produced when the sepiolite treatment was up to 5 %. The results showed that sepiolite-assisted remediation could potentially succeed on a field scale by decreasing Cd entry into the food chain.

  19. Long-term reactive nitrogen loading alters soil carbon and microbial community properties in a subalpine forest ecosystem

    Science.gov (United States)

    Boot, Claudia M.; Hall, Ed K.; Denef, Karolien; Baron, Jill S.

    2016-01-01

    Elevated nitrogen (N) deposition due to increased fossil fuel combustion and agricultural practices has altered global carbon (C) cycling. Additions of reactive N to N-limited environments are typically accompanied by increases in plant biomass. Soil C dynamics, however, have shown a range of different responses to the addition of reactive N that seem to be ecosystem dependent. We evaluated the effect of N amendments on biogeochemical characteristics and microbial responses of subalpine forest organic soils in order to develop a mechanistic understanding of how soils are affected by N amendments in subalpine ecosystems. We measured a suite of responses across three years (2011–2013) during two seasons (spring and fall). Following 17 years of N amendments, fertilized soils were more acidic (control mean 5.09, fertilized mean 4.68), and had lower %C (control mean 33.7% C, fertilized mean 29.8% C) and microbial biomass C by 22% relative to control plots. Shifts in biogeochemical properties in fertilized plots were associated with an altered microbial community driven by reduced arbuscular mycorrhizal (control mean 3.2 mol%, fertilized mean 2.5 mol%) and saprotrophic fungal groups (control mean 17.0 mol%, fertilized mean 15.2 mol%), as well as a decrease in N degrading microbial enzyme activity. Our results suggest that decreases in soil C in subalpine forests were in part driven by increased microbial degradation of soil organic matter and reduced inputs to soil organic matter in the form of microbial biomass.

  20. Microbial biomass and bacterial functional diversity in forest soils: effects of organic matter removal, compaction, and vegetation control

    Science.gov (United States)

    Qingchao Li; H. Lee Allen; Arthur G. Wollum

    2004-01-01

    The effects of organic matter removal, soil compaction, and vegetation control on soil microbial biomass carbon, nitrogen, C-to-N ratio, and functional diversity were examined in a 6-year loblolly pine plantation on a Coastal Plain site in eastern North Carolina, USA. This experimental plantation was established as part of the US Forest Service's Long Term Soil...

  1. Trace Metal Requirements for Microbial Enzymes Involved in the Production and Consumption of Methane and Nitrous Oxide

    Science.gov (United States)

    Glass, Jennifer B.; Orphan, Victoria J.

    2011-01-01

    Fluxes of greenhouse gases to the atmosphere are heavily influenced by microbiological activity. Microbial enzymes involved in the production and consumption of greenhouse gases often contain metal cofactors. While extensive research has examined the influence of Fe bioavailability on microbial CO2 cycling, fewer studies have explored metal requirements for microbial production and consumption of the second- and third-most abundant greenhouse gases, methane (CH4), and nitrous oxide (N2O). Here we review the current state of biochemical, physiological, and environmental research on transition metal requirements for microbial CH4 and N2O cycling. Methanogenic archaea require large amounts of Fe, Ni, and Co (and some Mo/W and Zn). Low bioavailability of Fe, Ni, and Co limits methanogenesis in pure and mixed cultures and environmental studies. Anaerobic methane oxidation by anaerobic methanotrophic archaea (ANME) likely occurs via reverse methanogenesis since ANME possess most of the enzymes in the methanogenic pathway. Aerobic CH4 oxidation uses Cu or Fe for the first step depending on Cu availability, and additional Fe, Cu, and Mo for later steps. N2O production via classical anaerobic denitrification is primarily Fe-based, whereas aerobic pathways (nitrifier denitrification and archaeal ammonia oxidation) require Cu in addition to, or possibly in place of, Fe. Genes encoding the Cu-containing N2O reductase, the only known enzyme capable of microbial N2O conversion to N2, have only been found in classical denitrifiers. Accumulation of N2O due to low Cu has been observed in pure cultures and a lake ecosystem, but not in marine systems. Future research is needed on metalloenzymes involved in the production of N2O by enrichment cultures of ammonia oxidizing archaea, biological mechanisms for scavenging scarce metals, and possible links between metal bioavailability and greenhouse gas fluxes in anaerobic environments where metals may be limiting due to sulfide

  2. DETERMINATION OF ENZYMES PRODUCED BY CERIPORIOPSIS SUBVERMISPORA DURING PRETREATMENT OF DIFFERENT BIOMASS SOURCES

    Directory of Open Access Journals (Sweden)

    Miroslav Ondrejovič

    2012-02-01

    Full Text Available The aim of this paper was to study of lignocellulolytic enzymes producing by Ceriporiopsis subvermispora during its cultivation on three types of plant biomass differentiated by chemical composition and physical properties (wheat straw, pine and poplar wood. The activity of lignocellulolytic enzymes in cultivation medium was determined by catalytic transformation of their natural substrates to products which were detected by photometric methods. Cellulase activities were very low while xylanases predominated. Wheat straw was best substrate for production of cellulases (4.38 U/mL and xylanases (23.34 U/mL. The maximum activity of cellulase and xylanase was reached at 8th and 3rd day, respectively. Laccase activity reached the maximum after 16 days and then gradually decreased. The best substrate for production of laccases was poplar wood (1.67 U/mL.

  3. Impact of phenazine-1-carboxylic acid upon iron speciation and microbial biomass in the rhizosphere of wheat

    Science.gov (United States)

    LeTourneau, M.; Marshall, M.; Grant, M.; Freeze, P.; Cliff, J. B.; Lai, B.; Strawn, D. G.; Thomashow, L. S.; Weller, D. M.; Harsh, J. B.

    2015-12-01

    Phenazine-1-carboxylic acid (PCA) is a redox-active antibiotic produced by diverse bacterial taxa, and has been shown to facilitate interactions between biofilms and iron (hydr)oxides in culture systems (Wang et al. 2011, J Bacteriol 192: 365). Because rhizobacterial biofilms are a major sink for plant-derived carbon and source for soil organic matter (SOM), and Fe (hydr)oxides have reactive surfaces that influence the stability of microbial biomass and SOM, PCA-producing rhizobacteria could influence soil carbon fluxes. Large populations of Pseudomonas fluorescens strains producing PCA in concentrations up to 1 μg/g root have been observed in the rhizosphere of non-irrigated wheat fields covering 1.56 million hectares of central Washington state. This is one of the highest concentrations ever reported for a natural antibiotic in a terrestrial ecosystem (Mavrodi et al. 2012, Appl Environ Microb 78: 804). Microscopic comparisons of PCA-producing (PCA+) and non-PCA-producing (PCA-) rhizobacterial colony morphologies, and comparisons of Fe extractions from rhizosphere soil inoculated with PCA+ and PCA- strains suggest that PCA promotes biofilm development as well as dramatic Fe transformations throughout the rhizosphere (unpublished data). In order to illustrate PCA-mediated interactions between biofilms and Fe (hydr)oxides in the rhizosphere, identify the specific Fe phases favored by PCA, and establish the ramifications for stability and distribution of microbial biomass and SOM, we have collected electron micrographs, X-ray fluorescence images, X-ray absorption near-edge spectra, and secondary-ion mass spectrometry images of wheat root sections inoculated with 15N-labelled PCA+ or PCA- rhizobacteria. These images and spectra allow us to assess the accumulation, turnover, and distribution of microbial biomass, the associations between Fe and other nutrients such as phosphorus, and the redox status and speciation of iron in the presence and absence of PCA. This

  4. Unravelling the Interactions between Hydrolytic and Oxidative Enzymes in Degradation of Lignocellulosic Biomass by Sporothrix carnis under Various Fermentation Conditions

    Directory of Open Access Journals (Sweden)

    Olusola A. Ogunyewo

    2016-01-01

    Full Text Available The mechanism underlying the action of lignocellulolytic enzymes in biodegradation of lignocellulosic biomass remains unclear; hence, it is crucial to investigate enzymatic interactions involved in the process. In this study, degradation of corn cob by Sporothrix carnis and involvement of lignocellulolytic enzymes in biodegradation were investigated over 240 h cultivation period. About 60% degradation of corn cob was achieved by S. carnis at the end of fermentation. The yields of hydrolytic enzymes, cellulase and xylanase, were higher than oxidative enzymes, laccase and peroxidase, over 144 h fermentation period. Maximum yields of cellulase (854.4 U/mg and xylanase (789.6 U/mg were at 96 and 144 h, respectively. Laccase and peroxidase were produced cooperatively with maximum yields of 489.06 U/mg and 585.39 U/mg at 144 h. Drastic decline in production of cellulase at 144 h (242.01 U/mg and xylanase at 192 h (192.2 U/mg indicates that they play initial roles in biodegradation of lignocellulosic biomass while laccase and peroxidase play later roles. Optimal degradation of corn cob (76.6% and production of hydrolytic and oxidative enzymes were achieved with 2.5% inoculum at pH 6.0. Results suggest synergy in interactions between the hydrolytic and oxidative enzymes which can be optimized for improved biodegradation.

  5. Two Strategies for Microbial Production of an Industrial Enzyme-Alpha-Amylase

    Science.gov (United States)

    Bernhardsdotter, Eva C. M. J.; Garriott, Owen; Pusey, Marc L.; Ng, Joseph D.

    2003-01-01

    Extremophiles are microorganisms that thrive in, from an anthropocentric view, extreme environments including hot springs, soda lakes and arctic water. This ability of survival at extreme conditions has rendered extremophiles to be of interest in astrobiology, evolutionary biology as well as in industrial applications. Of particular interest to the biotechnology industry are the biological catalysts of the extremophiles, the extremozymes, whose unique stabilities at extreme conditions make them potential sources of novel enzymes in industrial applications. There are two major approaches to microbial enzyme production. This entails enzyme isolation directly from the natural host or creating a recombinant expression system whereby the targeted enzyme can be overexpressed in a mesophilic host. We are employing both methods in the effort to produce alpha-amylases from a hyperthermophilic archaeon (Thermococcus) isolated from a hydrothermal vent in the Atlantic Ocean, as well as from alkaliphilic bacteria (Bacillus) isolated from a soda lake in Tanzania. Alpha-amylases catalyze the hydrolysis of internal alpha-1,4-glycosidic linkages in starch to produce smaller sugars. Thermostable alpha-amylases are used in the liquefaction of starch for production of fructose and glucose syrups, whereas alpha-amylases stable at high pH have potential as detergent additives. The alpha-amylase encoding gene from Thermococcus was PCR amplified using carefully designed primers and analyzed using bioinformatics tools such as BLAST and Multiple Sequence Alignment for cloning and expression in E.coli. Four strains of Bacillus were grown in alkaline starch-enriched medium of which the culture supernatant was used as enzyme source. Amylolytic activity was detected using the starch-iodine method.

  6. NRSA enzyme decomposition model data

    Data.gov (United States)

    U.S. Environmental Protection Agency — Microbial enzyme activities measured at more than 2000 US streams and rivers. These enzyme data were then used to predict organic matter decomposition and microbial...

  7. Anaerobic 4-hydroxyproline utilization: Discovery of a new glycyl radical enzyme in the human gut microbiome uncovers a widespread microbial metabolic activity.

    Science.gov (United States)

    Huang, Yolanda Y; Martínez-Del Campo, Ana; Balskus, Emily P

    2018-02-06

    The discovery of enzymes responsible for previously unappreciated microbial metabolic pathways furthers our understanding of host-microbe and microbe-microbe interactions. We recently identified and characterized a new gut microbial glycyl radical enzyme (GRE) responsible for anaerobic metabolism of trans-4-hydroxy-l-proline (Hyp). Hyp dehydratase (HypD) catalyzes the removal of water from Hyp to generate Δ 1 -pyrroline-5-carboxylate (P5C). This enzyme is encoded in the genomes of a diverse set of gut anaerobes and is prevalent and abundant in healthy human stool metagenomes. Here, we discuss the roles HypD may play in different microbial metabolic pathways as well as the potential implications of this activity for colonization resistance and pathogenesis within the human gut. Finally, we present evidence of anaerobic Hyp metabolism in sediments through enrichment culturing of Hyp-degrading bacteria, highlighting the wide distribution of this pathway in anoxic environments beyond the human gut.

  8. Cellulase enzyme and biomass utilization

    African Journals Online (AJOL)

    STORAGESEVER

    2009-06-03

    Jun 3, 2009 ... human population grows and economic development. However, the current .... conditions and the production cost of the related enzyme system. Therefore ... Given the importance of this enzyme to these so many industries,.

  9. Cultivation of high-biomass crops on coal mine spoil banks: Can microbial inoculation compensate for high doses of organic matter?

    Energy Technology Data Exchange (ETDEWEB)

    Gryndler, M.; Sudova, R.; Puschel, D.; Rydlova, J.; Janouskova, M.; Vosatka, M. [Academy of Science Czech Republic, Pruhonice (Czech Republic)

    2008-09-15

    Two greenhouse experiments were focused on the application of arbuscular mycorrhizal fungi (AMF) and plant growth promoting rhizobacteria (PGPR) in planting of high-biomass crops on reclaimed spoil banks. In the first experiment, we tested the effects of different organic amendments on growth of alfalfa and on the introduced microorganisms. While growth of plants was supported in substrate with compost amendment, mycorrhizal colonization was suppressed. Lignocellulose papermill waste had no negative effects on AMF, but did not positively affect growth of plants. The mixture of these two amendments was found to be optimal in both respects, plant growth and mycorrhizal development. Decreasing doses of this mixture amendment were used in the second experiment, where the effects of microbial inoculation (assumed to compensate for reduced doses of organic matter) on growth of two high-biomass crops, hemp and reed canarygrass, were studied. Plant growth response to microbial inoculation was either positive or negative, depending on the dose of the applied amendment and plant species.

  10. Seasonal and interannual dynamics of soil microbial biomass and available nitrogen in an alpine meadow in the eastern part of Qinghai-Tibet Plateau, China

    Science.gov (United States)

    Xu, Bo; Wang, Jinniu; Wu, Ning; Wu, Yan; Shi, Fusun

    2018-01-01

    Soil microbial activity varies seasonally in frozen alpine soils during cold seasons and plays a crucial role in available N pool accumulation in soil. The intra- and interannual patterns of microbial and nutrient dynamics reflect the influences of changing weather factors, and thus provide important insights into the biogeochemical cycles and ecological functions of ecosystems. We documented the seasonal and interannual dynamics of soil microbial and available N in an alpine meadow in the eastern part of Qinghai-Tibet Plateau, China, between April 2011 and October 2013. Soil was collected in the middle of each month and analyzed for water content, microbial biomass C (MBC) and N (MBN), dissolved organic C and N, and inorganic N. Soil microbial community composition was measured by the dilution-plate method. Fungi and actinomycetes dominated the microbial community during the nongrowing seasons, and the proportion of bacteria increased considerably during the early growing seasons. Trends of consistently increasing MBC and available N pools were observed during the nongrowing seasons. MBC sharply declined during soil thaw and was accompanied by a peak in available N pool. Induced by changes in soil temperatures, significant shifts in the structures and functions of microbial communities were observed during the winter-spring transition and largely contributed to microbial reduction. The divergent seasonal dynamics of different N forms showed a complementary nutrient supply pattern during the growing season. Similarities between the interannual dynamics of microbial biomass and available N pools were observed, and soil temperature and water conditions were the primary environmental factors driving interannual fluctuations. Owing to the changes in climate, seasonal soil microbial activities and nutrient supply patterns are expected to change further, and these changes may have crucial implications for the productivity and biodiversity of alpine ecosystems.

  11. Lignin from hydrothermally pretreated grass biomass retards enzymatic cellulose degradation by acting as a physical barrier rather than by inducing nonproductive adsorption of enzymes.

    Science.gov (United States)

    Djajadi, Demi T; Jensen, Mads M; Oliveira, Marlene; Jensen, Anders; Thygesen, Lisbeth G; Pinelo, Manuel; Glasius, Marianne; Jørgensen, Henning; Meyer, Anne S

    2018-01-01

    Lignin is known to hinder efficient enzymatic conversion of lignocellulose in biorefining processes. In particular, nonproductive adsorption of cellulases onto lignin is considered a key mechanism to explain how lignin retards enzymatic cellulose conversion in extended reactions. Lignin-rich residues (LRRs) were prepared via extensive enzymatic cellulose degradation of corn stover ( Zea mays subsp. mays L.), Miscanthus  ×  giganteus stalks (MS) and wheat straw ( Triticum aestivum L.) (WS) samples that each had been hydrothermally pretreated at three severity factors (log R 0 ) of 3.65, 3.83 and 3.97. The LRRs had different residual carbohydrate levels-the highest in MS; the lowest in WS. The residual carbohydrate was not traceable at the surface of the LRRs particles by ATR-FTIR analysis. The chemical properties of the lignin in the LRRs varied across the three types of biomass, but monolignols composition was not affected by the severity factor. When pure cellulose was added to a mixture of LRRs and a commercial cellulolytic enzyme preparation, the rate and extent of glucose release were unaffected by the presence of LRRs regardless of biomass type and severity factor, despite adsorption of the enzymes to the LRRs. Since the surface of the LRRs particles were covered by lignin, the data suggest that the retardation of enzymatic cellulose degradation during extended reaction on lignocellulosic substrates is due to physical blockage of the access of enzymes to the cellulose caused by the gradual accumulation of lignin at the surface of the biomass particles rather than by nonproductive enzyme adsorption. The study suggests that lignin from hydrothermally pretreated grass biomass retards enzymatic cellulose degradation by acting as a physical barrier blocking the access of enzymes to cellulose rather than by inducing retardation through nonproductive adsorption of enzymes.

  12. Effect of dry mycelium of Penicillium chrysogenum fertilizer on soil microbial community composition, enzyme activities and snap bean growth.

    Science.gov (United States)

    Wang, Bing; Liu, Huiling; Cai, Chen; Thabit, Mohamed; Wang, Pu; Li, Guomin; Duan, Ziheng

    2016-10-01

    The dry mycelium fertilizer (DMF) was produced from penicillin fermentation fungi mycelium (PFFM) following an acid-heating pretreatment to degrade the residual penicillin. In this study, it was applied into soil as fertilizer to investigate its effects on soil properties, phytotoxicity, microbial community composition, enzyme activities, and growth of snap bean in greenhouse. As the results show, pH, total nitrogen, total phosphorus, total potassium, and organic matter of soil with DMF treatments were generally higher than CON treatment. In addition, the applied DMF did not cause heavy metal and residual drug pollution of the modified soil. The lowest GI values (<0.3) were recorded at DMF8 (36 kg DMF/plat) on the first days after applying the fertilizer, indicating that severe phytotoxicity appeared in the DMF8-modified soil. Results of microbial population and enzyme activities illustrated that DMF was rapidly decomposed and the decomposition process significantly affected microbial growth and enzyme activities. The DMF-modified soil phytotoxicity decreased at the late fertilization time. DMF1 was considered as the optimum amount of DMF dose based on principal component analysis scores. Plant height and plant yield of snap bean were remarkably enhanced with the optimum DMF dose.

  13. Effect of 40 and 80 Years of Conifer Regrowth on Soil Microbial Activities and Community Structure in Subtropical Low Mountain Forests

    Directory of Open Access Journals (Sweden)

    Ed-Haun Chang

    2016-10-01

    Full Text Available The effects of long-term reforestation on soil microbial communities and biomass are poorly understood. This study was conducted on two coniferous plantations: Cunninghamia konishii Hayata, planted 40 years ago (CONIF-40, and Calocedrus formosana (Florin Florin, planted 80 years ago (CONIF-80. An adjacent natural broadleaf forest (BROAD-Nat was used as a control. We determined microbial biomass C and N contents, enzyme activities, and community composition (via phospholipid fatty acid [PLFA] assessment. Both microbial biomass and PLFA content were higher in the summer than in the winter and differed among the forests in summer only. Total PLFA, total bacterial, gram-positive bacterial, gram-negative bacterial, and vesicular arbuscular mycorrhizal fungal contents followed the same pattern. Total fungal content and the ratios of fungi to bacteria and of gram-positive to gram-negative bacteria were highest in CONIF-40, with no difference between the other forests. Principal component analysis of PLFA contents revealed that CONIF-40 communities were distinct from those of CONIF-80 and BROAD-Nat. Our results suggest that vegetation replacement during reforestation exerts a prolonged impact on the soil microbial community. The understory broadleaf shrubs and trees established after coniferous plantation reforestation may balance out the effects of coniferous litter, contributing to bacterial recovery.

  14. The combined effect of decabromodiphenyl ether (BDE-209) and copper (Cu) on soil enzyme activities and microbial community structure.

    Science.gov (United States)

    Zhang, Wei; Zhang, Meng; An, Shuai; Lin, Kuangfei; Li, Hui; Cui, Changzheng; Fu, Rongbing; Zhu, Jiang

    2012-09-01

    Waste electrical and electronic equipment (e-waste) is now the fastest growing waste stream in the world. It is reported that polybrominated diphenyl ethers (PBDEs) and heavy metals were main contaminants in e-waste recycling site. Among these contaminants BDE-209 and Cu were widespread, yet their combined effect on soil enzyme activities and microbial community structure are not well understood. In this study, the ecotoxicological effects of both combined and single pollution of BDE-209 and Cu at different concentration levels were studied under laboratory conditions. The activities of soil catalase, urease and saccharase were sensitive to BDE-209 and Cu pollution. Although the enzyme activities varied over time, the concentration effects were obvious. Statistical analyses revealed that, at the same incubation time, as the concentration of BDE-209 or Cu increased, the enzyme activities were decreased. Combined effects of both BDE-209 and Cu were different from that of BDE-209 or Cu alone. Enzyme activities data were essentially based on the multiple regression technique. The results showed that the action and interaction between BDE-209 and Cu were strongly dependent on the exposure time, as the combined effects of BDE-209 and Cu were either synergistic or antagonistic at different incubation times. Soil catalase and saccharase were more comfortable used as indicators of BDE-209 and Cu combined pollution, as the variation trends were similar to the single contaminant treatments, and the responses were quick and significant. Denaturing Gradient Gel Electrophoresis (DGGE) analysis of bacterial 16S rDNA gene showed that BDE-209 and Cu pollution altered the bacterial community structure by promoting changes in species composition and species richness. The existence of BDE-209 and Cu in soils reduced the microbial diversity, and the concentration effects were obvious. Overall, microbial diversity in the combined treatments were lower than the single ones, and when the

  15. Characterization of Cellulase Enzyme Inhibitors Formed During the Chemical Pretreatments of Rice Straw

    Science.gov (United States)

    Rajan, Kalavathy

    Production of fuels and chemicals from a renewable and inexpensive resource such as lignocellulosic biomass is a lucrative and sustainable option for the advanced biofuel and bio-based chemical platform. Agricultural residues constitute the bulk of potential feedstock available for cellulosic fuel production. On a global scale, rice straw is the largest source of agricultural residues and is therefore an ideal crop model for biomass deconstruction studies. Lignocellulosic biofuel production involves the processes of biomass conditioning, enzymatic saccharification, microbial fermentation and ethanol distillation, and one of the major factors affecting its techno-economic feasibility is the biomass recalcitrance to enzymatic saccharification. Preconditioning of lignocellulosic biomass, using chemical, physico-chemical, mechanical and biological pretreatments, is often practiced such that biomass becomes available to downstream processing. Pretreatments, such as dilute acid and hot water, are effective means of biomass conversion. However, despite their processing importance, preconditioning biomass also results in the production of carbohydrate and lignin degradation products that are inhibitory to downstream saccharification enzymes. The saccharification enzyme cocktail is made up of endo-cellulase, exo-cellulase and beta-glucosidase enzymes, whose role is to cleave cellulose polymers into glucose monomers. Specifically, endo-cellulase and exo-cellulase enzymes cleave cellulose chains in the middle and at the end, resulting in cellobiose molecules, which are hydrolyzed into glucose by beta-glucosidase. Unfortunately, degradation compounds generated during pretreatment inhibit the saccharification enzyme cocktail. Various research groups have identified specific classes of inhibitors formed during biomass pretreatment and have studied their inhibitory effect on the saccharification cocktail. These various research groups prepared surrogate solutions in an attempt to

  16. Microbial Indicators of Soil Quality under Different Land Use Systems in Subtropical Soils

    Science.gov (United States)

    Maharjan, M.

    2016-12-01

    Land-use change from native forest to intensive agricultural systems can negatively impact numerous soil parameters. Understanding the effects of forest ecosystem transformations on markers of long-term soil health is particularly important in rapidly developing regions such as Nepal, where unprecedented levels of agriculturally-driven deforestation have occurred in recent decades. However, the effects of widespread land use changes on soil quality in this region have yet to be properly characterized. Microbial indicators (soil microbial biomass, metabolic quotient and enzymes activities) are particularly suited to assessing the consequences of such ecosystem disturbances, as microbial communities are especially sensitive to environmental change. Thus, the aim of this study was to assess the effect of land use system; i.e. forest, organic and conventional farming, on soil quality in Chitwan, Nepal using markers of microbial community size and activity. Total organic C and N contents were higher in organic farming compared with conventional farming and forest, suggesting higher nutrient retention and soil preservation with organic farming practices compared to conventional. These differences in soil composition were reflected in the health of the soil microbial communities: Organic farm soil exhibited higher microbial biomass C, elevated β-glucosidase and chitinase activities, and a lower metabolic quotient relative to other soils, indicating a larger, more active, and less stressed microbial community, respectively. These results collectively demonstrate that application of organic fertilizers and organic residues positively influence nutrient availability, with subsequent improvements in soil quality and productivity. Furthermore, the sensitivity of microbial indicators to different management practices demonstrated in this study supports their use as effective markers of ecosystem disturbance in subtropical soils.

  17. Spatial distribution of microbial biomass, activity, community structure, and the biodegradation of linear alkylbenzene sulfonate (LAS) and linear alcohol ethoxylate (LAE) in the subsurface.

    Science.gov (United States)

    Federle, T W; Ventullo, R M; White, D C

    1990-12-01

    The vertical distribution of microbial biomass, activity, community structure and the mineralization of xenobiotic chemicals was examined in two soil profiles in northern Wisconsin. One profile was impacted by infiltrating wastewater from a laundromat, while the other served as a control. An unconfined aquifer was present 14 meters below the surface at both sites. Biomass and community structure were determined by acridine orange direct counts and measuring concentrations of phospholipid-derived fatty acids (PLFA). Microbial activity was estimated by measuring fluorescein diacetate (FDA) hydrolysis, thymidine incorporation into DNA, and mixed amino acid (MAA) mineralization. Mineralization kinetics of linear alkylbenzene sulfonate (LAS) and linear alcohol ethoxylate (LAE) were determined at each depth. Except for MAA mineralization rates, measures of microbial biomass and activity exhibited similar patterns with depth. PLFA concentration and rates of FDA hydrolysis and thymidine incorporation decreased 10-100 fold below 3 m and then exhibited little variation with depth. Fungal fatty acid markers were found at all depths and represented from 1 to 15% of the total PLFAs. The relative proportion of tuberculostearic acid (TBS), an actinomycete marker, declined with depth and was not detected in the saturated zone. The profile impacted by wastewater exhibited higher levels of PLFA but a lower proportion of TBS than the control profile. This profile also exhibited faster rates of FDA hydrolysis and amino acid mineralization at most depths. LAS was mineralized in the upper 2 m of the vadose zone and in the saturated zone of both profiles. Little or no LAS biodegradation occurred at depths between 2 and 14 m. LAE was mineralized at all depths in both profiles, and the mineralization rate exhibited a similar pattern with depth as biomass and activity measurements. In general, biomass and biodegradative activities were much lower in groundwater than in soil samples obtained

  18. Microbial Tyrosinases: Promising Enzymes for Pharmaceutical, Food Bioprocessing, and Environmental Industry

    Directory of Open Access Journals (Sweden)

    Kamal Uddin Zaidi

    2014-01-01

    Full Text Available Tyrosinase is a natural enzyme and is often purified to only a low degree and it is involved in a variety of functions which mainly catalyse the o-hydroxylation of monophenols into their corresponding o-diphenols and the oxidation of o-diphenols to o-quinones using molecular oxygen, which then polymerizes to form brown or black pigments. The synthesis of o-diphenols is a potentially valuable catalytic ability and thus tyrosinase has attracted a lot of attention with respect to industrial applications. In environmental technology it is used for the detoxification of phenol-containing wastewaters and contaminated soils, as biosensors for phenol monitoring, and for the production of L-DOPA in pharmaceutical industries, and is also used in cosmetic and food industries as important catalytic enzyme. Melanin pigment synthesized by tyrosinase has found applications for protection against radiation cation exchangers, drug carriers, antioxidants, antiviral agents, or immunogen. The recombinant V. spinosum tryosinase protein can be used to produce tailor-made melanin and other polyphenolic materials using various phenols and catechols as starting materials. This review compiles the recent data on biochemical and molecular properties of microbial tyrosinases, underlining their importance in the industrial use of these enzymes. After that, their most promising applications in pharmaceutical, food processing, and environmental fields are presented.

  19. Heterologous expression of xylanase enzymes in lipogenic yeast Yarrowia lipolytica.

    Directory of Open Access Journals (Sweden)

    Wei Wang

    Full Text Available To develop a direct microbial sugar conversion platform for the production of lipids, drop-in fuels and chemicals from cellulosic biomass substrate, we chose Yarrowia lipolytica as a viable demonstration strain. Y. lipolytica is known to accumulate lipids intracellularly and is capable of metabolizing sugars to produce lipids; however, it lacks the lignocellulose-degrading enzymes needed to break down biomass directly. While research is continuing on the development of a Y. lipolytica strain able to degrade cellulose, in this study, we present successful expression of several xylanases in Y. lipolytica. The XynII and XlnD expressing Yarrowia strains exhibited an ability to grow on xylan mineral plates. This was shown by Congo Red staining of halo zones on xylan mineral plates. Enzymatic activity tests further demonstrated active expression of XynII and XlnD in Y. lipolytica. Furthermore, synergistic action in converting xylan to xylose was observed when XlnD acted in concert with XynII. The successful expression of these xylanases in Yarrowia further advances us toward our goal to develop a direct microbial conversion process using this organism.

  20. Integrated cellulosic enzymes hydrolysis and fermentative advanced yeast bioconversion solution ready for biomass biorefineries

    Energy Technology Data Exchange (ETDEWEB)

    Kumar, Manoj [DSM Innovation, Inc., San Francisco, CA (United States)

    2011-05-04

    These are slides from this conference. Lignocellulosic biomass is the most abundant, least expensive renewable natural biological resource for the production of biobased products and bioenergy is important for the sustainable development of human civilization in 21st century. For making the fermentable sugars from lignocellulosic biomass, a reduction in cellulase production cost, an improvement in cellulase performance, and an increase in sugar yields are all vital to reduce the processing costs of biorefineries. Improvements in specific cellulase activities for non-complexed cellulase mixtures can be implemented through cellulase engineering based on rational design or directed evolution for each cellulase component enzyme, as well as on the reconstitution of cellulase components. In this paper, we will provide DSM's efforts in cellulase research and developments and focus on limitations. Cellulase improvement strategies based on directed evolution using screening on relevant substrates, screening for higher thermal tolerance based on activity screening approaches such as continuous culture using insoluble cellulosic substrates as a powerful selection tool for enriching beneficial cellulase mutants from the large library. We will illustrate why and how thermostable cellulases are vital for economic delivery of bioproducts from cellulosic biomass using biochemical conversion approach.

  1. Metagenomic Analysis of the Gut Microbiome of the Common Black Slug Arion ater in Search of Novel Lignocellulose Degrading Enzymes

    Directory of Open Access Journals (Sweden)

    Ryan Joynson

    2017-11-01

    Full Text Available Some eukaryotes are able to gain access to well-protected carbon sources in plant biomass by exploiting microorganisms in the environment or harbored in their digestive system. One is the land pulmonate Arion ater, which takes advantage of a gut microbial consortium that can break down the widely available, but difficult to digest, carbohydrate polymers in lignocellulose, enabling them to digest a broad range of fresh and partially degraded plant material efficiently. This ability is considered one of the major factors that have enabled A. ater to become one of the most widespread plant pest species in Western Europe and North America. Using metagenomic techniques we have characterized the bacterial diversity and functional capability of the gut microbiome of this notorious agricultural pest. Analysis of gut metagenomic community sequences identified abundant populations of known lignocellulose-degrading bacteria, along with well-characterized bacterial plant pathogens. This also revealed a repertoire of more than 3,383 carbohydrate active enzymes (CAZymes including multiple enzymes associated with lignin degradation, demonstrating a microbial consortium capable of degradation of all components of lignocellulose. This would allow A. ater to make extensive use of plant biomass as a source of nutrients through exploitation of the enzymatic capabilities of the gut microbial consortia. From this metagenome assembly we also demonstrate the successful amplification of multiple predicted gene sequences from metagenomic DNA subjected to whole genome amplification and expression of functional proteins, facilitating the low cost acquisition and biochemical testing of the many thousands of novel genes identified in metagenomics studies. These findings demonstrate the importance of studying Gastropod microbial communities. Firstly, with respect to understanding links between feeding and evolutionary success and, secondly, as sources of novel enzymes with

  2. Subtropical urban turfs: Carbon and nitrogen pools and the role of enzyme activity.

    Science.gov (United States)

    Kong, Ling; Chu, L M

    2018-03-01

    Urban grasslands not only provide a recreational venue for urban residents, but also sequester organic carbon in vegetation and soils through photosynthesis, and release carbon dioxide through respiration, which largely contribute to carbon storage and fluxes at regional and global scales. We investigated organic carbon and nitrogen pools in subtropical turfs and found that dissolved organic carbon (DOC) and dissolved organic nitrogen (DON) were regulated by several factors including microbial activity which is indicated by soil enzymatic activity. We observed a vertical variation and different temporal patterns in both soil DOC, DON and enzyme activities, which decreased significantly with increasing soil depths. We further found that concentration of soil DON was linked with turf age. There were correlations between grass biomass and soil properties, and soil enzyme activities. In particular, soil bulk density was significantly correlated with soil moisture and soil organic carbon (SOC). In addition, DOC correlated significantly with DON. Significant negative correlations were also observed between soil total dissolved nitrogen (TDN) and grass biomass of Axonopus compressus and Zoysia matrella. Specifically, grass biomass was significantly correlated with the soil activity of urease and β-glucosidase. Soil NO 3 -N concentration also showed negative correlations with the activity of both β-glucosidase and protease but there were no significant correlations between cellulase and soil properties or grass biomass. Our study demonstrated a relationship between soil C and N dynamics and soil enzymes that could be modulated to enhance SOC pools through management and maintenance practices. Copyright © 2017. Published by Elsevier B.V.

  3. The putative endoglucanase PcGH61D from Phanerochaete chrysosporium is a metal-dependent oxidative enzyme that cleaves cellulose.

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    Bjørge Westereng

    Full Text Available Many fungi growing on plant biomass produce proteins currently classified as glycoside hydrolase family 61 (GH61, some of which are known to act synergistically with cellulases. In this study we show that PcGH61D, the gene product of an open reading frame in the genome of Phanerochaete chrysosporium, is an enzyme that cleaves cellulose using a metal-dependent oxidative mechanism that leads to generation of aldonic acids. The activity of this enzyme and its beneficial effect on the efficiency of classical cellulases are stimulated by the presence of electron donors. Experiments with reduced cellulose confirmed the oxidative nature of the reaction catalyzed by PcGH61D and indicated that the enzyme may be capable of penetrating into the substrate. Considering the abundance of GH61-encoding genes in fungi and genes encoding their functional bacterial homologues currently classified as carbohydrate binding modules family 33 (CBM33, this enzyme activity is likely to turn out as a major determinant of microbial biomass-degrading efficiency.

  4. Biomassa e atividade microbianas do solo sob influência de chumbo e da rizosfera da soja micorrizada Soil microbial biomass and activity under the influence of lead addition and mycorrhizal soybean rhizosphere

    Directory of Open Access Journals (Sweden)

    Sara Adrián López de Andrade

    2004-12-01

    Full Text Available O objetivo deste trabalho foi avaliar o efeito da adição de chumbo (Pb ao solo na biomassa e atividade microbianas do solo sob influência da rizosfera de soja micorrizada. O trabalho foi realizado em casa de vegetação, com delineamento inteiramente casualizado num esquema fatorial 4x2x2 utilizando-se 0, 150, 300 e 600 mg dm-3 de Pb, inoculação ou não do fungo micorrízico arbuscular (FMA, Glomus macrocarpum, e duas épocas de amostragem - florescimento e maturação da soja. Avaliaram-se o C da biomassa microbiana, a liberação de CO2 do solo e a atividade de três enzimas, desidrogenase, fosfatase alcalina e arilssulfatase. O Pb afetou negativamente o C da biomassa e a atividade da microbiota rizosférica, ocorrendo interação entre a presença de propágulos de FMA e o estádio de desenvolvimento da planta. A atividade da fosfatase alcalina foi a mais afetada pelas altas concentrações de Pb adicionadas ao solo, com redução de 60% na sua atividade, mostrando-se um indicador sensível do estresse metabólico da comunidade microbiana do solo causado pelo excesso de chumbo. A micorrização da soja influenciou de forma direta a microbiota rizosférica, resultando em maior atividade e biomassa, principalmente no estádio de maturação da soja. A microbiota do solo apresentou sintomas de estresse decorrentes da adição de chumbo.The objective of this work was to evaluate the effects of lead addition on soil microbial biomass and activity under the influence of the rhizosphere of mycorrhizal soybean. The experimental design was completely randomized and arranged in a 4x2x2 factorial scheme, using 0, 150, 300 and 600 mg dm-3, inoculation or not of the arbuscular mycorrhizal fungus (AMF Glomus macrocarpum and two sampling periods: soybean flowering and maturity. Microbial biomass C, soil respiration and the activity of three soil enzymes (deshydrogenase, alkaline phosphatase and arilsulphatase were determined. The most affected enzyme

  5. Functional analysis of the Glucan Degradation Locus (GDL) in Caldicellulosiruptor bescii reveals essential roles of component glycoside hydrolases in plant biomass deconstruction.

    Science.gov (United States)

    Conway, Jonathan M; McKinley, Bennett S; Seals, Nathaniel L; Hernandez, Diana; Khatibi, Piyum A; Poudel, Suresh; Giannone, Richard J; Hettich, Robert L; Williams-Rhaesa, Amanda M; Lipscomb, Gina L; Adams, Michael W W; Kelly, Robert M

    2017-10-06

    The ability to hydrolyze microcrystalline cellulose is an uncommon feature in the microbial world, but one that can be exploited for conversion of lignocellulosic feedstocks into bio-based fuels and chemicals. Understanding the physiological and biochemical mechanisms by which microorganisms deconstruct cellulosic material is key to achieving this objective. The Glucan Degradation Locus (GDL) in the genomes of extremely thermophilic Caldicellulosiruptor species encodes polysaccharide lyases (PLs), unique cellulose binding proteins (tāpirins), and putative post-translational modifying enzymes, in addition to multi-domain, multi-functional glycoside hydrolases (GHs), thereby representing an alternative paradigm for plant biomass degradation, as compared to fungal or cellulosomal systems. To examine the individual and collective in vivo roles of the glycolytic enzymes, the six GHs in the GDL of Caldicellulosiruptor bescii were systematically deleted, and the extent to which the resulting mutant strains could solubilize microcrystalline cellulose (Avicel) and plant biomasses (switchgrass or poplar) was examined. Three of the GDL enzymes, Athe_1867 (CelA) (GH9-CBM3-CBM3-CBM3-GH48), Athe_1859 (GH5-CBM3-CBM3-GH44), and Athe_1857 (GH10-CBM3-CBM3-GH48), acted synergistically in vivo and accounted for 92% of naked microcellulose (Avicel) degradation. However, the relative importance of the GDL GHs varied for the plant biomass substrates tested. Furthermore, mixed cultures of mutant strains showed switchgrass solubilization depended on the secretome-bound enzymes collectively produced by the culture and not on the specific strain from which they came. These results demonstrate that certain GDL GHs are primarily responsible for the degradation of microcrystalline-containing substrates by C. bescii and provide new insights into the workings of a novel microbial mechanism for lignocellulose utilization. Importance The efficient and extensive degradation of complex

  6. Soil carbon fractions and enzyme activities under different vegetation types on the Loess Plateau of China

    OpenAIRE

    Zhang, Haixin; Zeng, Quanchao; An, Shaoshan; Dong, Yanghong; Darboux, Frédéric

    2016-01-01

    Vegetation restoration was effective way of protecting soil erosion and water conservation on the Loess Plateau. Carbon fractions and enzyme activities were sensitive parameters for assessment of soil remediation through revegetation. Forest, forest steppe and grassland soils were collected at 0–5 cm and 5–20 cm soil layers in Yanhe watershed, Shaanxi Province. Urease, sucrase, alkaline phosphatase, soil organic carbon (SOC), microbial biomass carbon (MBC), easily ox...

  7. Biomass production and secretion of hydrolytic enzymes are influenced by the structural complexity of the nitrogen source in Fusarium oxysporum and Aspergillus nidulans.

    Science.gov (United States)

    da Silva, M C; Bertolini, M C; Ernandes, J R

    2001-01-01

    The structural complexity of the nitrogen sources strongly affects biomass production and secretion of hydrolytic enzymes in filamentous fungi. Fusarium oxysporum and Aspergillus nidulans were grown in media containing glucose or starch, and supplemented with a nitrogen source varying from a single ammonium salt (ammonium sulfate) to free amino acids (casamino acids), peptides (peptone) and protein (gelatin). In glucose, when the initial pH was adjusted to 5.0, for both microorganisms, higher biomass production occurred upon supplementation with a nitrogen source in the peptide form (peptone and gelatin). With a close to neutrality pH, biomass accumulation was lower only in the presence of the ammonium salt. When grown in starch, biomass accumulation and secretion of hydrolytic enzymes (amylolytic and proteolytic) by Fusarium also depended on the nature of the nitrogen supplement and the pH. When the initial pH was adjusted to 5.0, higher growth and higher amylolytic activities were detected in the media supplemented with peptone, gelatin and casamino acids. However, at pH 7.0, higher biomass accumulation and higher amylolytic activities were observed upon supplementation with peptone or gelatin. Ammonium sulfate and casamino acids induced a lower production of biomass, and a different level of amylolytic enzyme secretion: high in ammonium sulfate and low in casamino acids. Secretion of proteolytic activity was always higher in the media supplemented with peptone and gelatin. Aspergillus, when grown in starch, was not as dependent as Fusarium on the nature of nitrogen source or the pH. The results described in this work indicate that the metabolism of fungi is regulated not only by pH, but also by the level of structural complexity of the nitrogen source in correlation to the carbon source.

  8. In situ phytoremediation of PAH-contaminated soil by intercropping alfalfa (Medicago sativa L.) with tall fescue (Festuca arundinacea Schreb.) and associated soil microbial activity

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Mingming; Fu, Dengqiang; Teng, Ying; Shen, Yuanyuan; Luo, Yongming; Li, Zhengao [Chinese Academy of Sciences, Nanjing (China). Key Laboratory of Soil Environment and Pollution Remediation; Christie, Peter [Agri-Food and Biosciences Institute, Belfast (United Kingdom). Agri-Environment Branch

    2011-09-15

    Purpose: A 7-month field experiment was conducted to investigate the polycyclic aromatic hydrocarbon (PAH) remediation potential of two plant species and changes in counts of soil PAH-degrading bacteria and microbial activity. Materials and methods: Alfalfa and tall fescue were grown in monoculture and intercropped for 7 months in contaminated field soil. Soil and plant samples were analyzed for PAHs. Plant biomass, densities of PAH-degradation soil bacteria, soil microbial biomass C and N, enzyme activities, and the physiological profile of the soil microbial community were determined. Results and discussion: Average removal percentage of total PAHs in intercropping (30.5%) was significantly higher than in monoculture (19.9%) or unplanted soil (-0.6%). About 7.5% of 3-ring, 12.3% of 4-ring, and 17.2% of 5(+6)-ring PAHs were removed from the soil by alfalfa, with corresponding values of 25.1%, 10.4%, and 30.1% for tall fescue. Intercropping significantly enhanced the remediation efficiency. About 18.9% of 3-ring, 30.9% of 4-ring, and 33.4% of 5(+6)-ring PAHs were removed by the intercropping system. Higher counts of soil culturable PAH-degrading bacteria and elevated microbial biomass and enzyme activities were found after intercropping. Soil from intercropping showed significantly higher (p < 0.05) average well-color development obtained by the BIOLOG Ecoplate assay and Shannon-Weaver index compared with monoculture. Conclusions: Cropping promoted the dissipation of soil PAHs. Tall fescue gave greater removal of soil PAHs than alfalfa, and intercropping was more effective than monoculture. Intercropping of alfalfa and tall fescue may be a promising in situ bioremediation strategy for PAH-contaminated soils. (orig.)

  9. Soil microbial properties after long-term swine slurry application to conventional and no-tillage systems in Brazil.

    Science.gov (United States)

    Balota, Elcio L; Machineski, Oswaldo; Hamid, Karima I A; Yada, Ines F U; Barbosa, Graziela M C; Nakatani, Andre S; Coyne, Mark S

    2014-08-15

    Swine waste can be used as an agricultural fertilizer, but large amounts may accumulate excess nutrients in soil or contaminate the surrounding environment. This study evaluated long-term soil amendment (15 years) with different levels of swine slurry to conventional (plow) tillage (CT) and no tillage (NT) soils. Long-term swine slurry application did not affect soil organic carbon. Some chemical properties, such as calcium, base saturation, and aluminum saturation were significantly different within and between tillages for various application rates. Available P and microbial parameters were significantly affected by slurry addition. Depending on tillage, soil microbial biomass and enzyme activity increased up to 120 m(3) ha(-1) year(-1) in all application rates. The NT system had higher microbial biomass and activity than CT at all application levels. There was an inverse relationship between the metabolic quotient (qCO2) and MBC, and the qCO2 was 53% lower in NT than CT. Swine slurry increased overall acid phosphatase activity, but the phosphatase produced per unit of microbial biomass decreased. A comparison of data obtained in the 3rd and 15th years of swine slurry application indicated that despite slurry application the CT system degraded with time while the NT system had improved values of soil quality indicators. For these Brazilian oxisols, swine slurry amendment was insufficient to maintain soil quality parameters in annual crop production without additional changes in tillage management. Copyright © 2014 Elsevier B.V. All rights reserved.

  10. The effects of environmental physical factors on the microbial communities and the distribution of different CO2 fixation pathways in a limestone landscape

    Science.gov (United States)

    Wun, S. R.; Huang, T. Y.; Hsu, B. M.; Fan, C. W.

    2017-12-01

    We aimed to study the effects of physical factors on the relative abundance of bacteria and their preferential admissions of autotrophic CO2 fixation pathways after subjected to environmental long-term influence. The Narrow-Sky located in upper part of Takangshan is a small gulch of Pleistocene coralline limestone formation in southern Taiwan. The physical parameters such as illumination, humidity, and temperature were varied largely in habitats around the gulch, namely on the limestone wall at the opening of gulch, on the coordinate ground soil, on the wall inside the gulch, and the water drip from limestone wall. The total organic carbon was measured in solid samples to evaluate the biomass of the habitats. A metagenomic approach was carried out to reveal their microbial community structure. After the metagenomic library of operational taxonomic units (OTUs) was constructed, a BLAST search by "nomenclature of bacteria" instead of sequences between the OTU libraries and KEGG database was carried out to generate libraries of "model microbial communities", which the complete genomes of the entire bacterial populations were available. Our results showed the biomass of habitats in the opening of gulch was twice higher than the inside, suggesting the illumination played an important role in biosynthesis. In quantitative comparison in key enzymes of CO2 fixation pathways by model communities, 70% to 90% of bacteria possessed key enzymes of Fuchs-Holo cycle, while only 5% to 20% of bacteria contained key enzymes of Calvin-Benson cycle. The key enzymes for hydroxypropionate/ hydroxybutyrate and dicarboxylate/ 4-hydroxybutyrate cycles were not found in this study. In the water sample, approximate 10% of bacteria consisted of the key enzyme for Arnon-Buchanan cycle. Less than 2% of bacteria in all habitats take the reductive acetyl-CoA cycle for CO2 fixation. This study provides a novel method to study biosynthetic process of microbial communities in natural habitats.

  11. The Influence of Tallow on Rumen Metabolism, Microbial Biomass Synthesis and Fatty Acid Composition of Bacteria and Protozoa

    DEFF Research Database (Denmark)

    Weisbjerg, Martin Riis; Børsting, Christian Friis; Hvelplund, Torben

    1992-01-01

    Rumen metabolism, microbial biomass synthesis and microbial long chain fatty acid composition were studied in lactating cows fed at two levels of dry matter intake (L, 8.6 kg DM and H, 12.6 kg DM) with 0, 4 and 6% added tallow at the low feed level (L0, L4 and L6) and 0, 2, 4 and 6% at the high...... feed level (H0, H2, H4 and H6). Fibre digestibility was not significantly affected by tallow addition. Increasing tallow level in the diet decreased the total VFA concentration, the ratio of acetic acid to propionic acid and the ammonia concentration in the rumen. Crude fat and fatty acid content...... in bacterial and protozoal dry matter increased with increased tallow level, especially due to an increase in fatty acids originating from the feeds. Microbial synthesis in the rumen and flow of amino acids to the duodenum was highest for medium fat intake at the high feed level....

  12. Effect of elevated CO2 on chlorpyriphos degradation and soil microbial activities in tropical rice soil.

    Science.gov (United States)

    Adak, Totan; Munda, Sushmita; Kumar, Upendra; Berliner, J; Pokhare, Somnath S; Jambhulkar, N N; Jena, M

    2016-02-01

    Impact of elevated CO2 on chlorpyriphos degradation, microbial biomass carbon, and enzymatic activities in rice soil was investigated. Rice (variety Naveen, Indica type) was grown under four conditions, namely, chambered control, elevated CO2 (550 ppm), elevated CO2 (700 ppm) in open-top chambers and open field. Chlorpyriphos was sprayed at 500 g a.i. ha(-1) at maximum tillering stage. Chlorpyriphos degraded rapidly from rice soils, and 88.4% of initially applied chlorpyriphos was lost from the rice soil maintained under elevated CO2 (700 ppm) by day 5 of spray, whereas the loss was 80.7% from open field rice soil. Half-life values of chlorpyriphos under different conditions ranged from 2.4 to 1.7 days with minimum half-life recorded with two elevated CO2 treatments. Increased CO2 concentration led to increase in temperature (1.2 to 1.8 °C) that played a critical role in chlorpyriphos persistence. Microbial biomass carbon and soil enzymatic activities specifically, dehydrogenase, fluorescien diacetate hydrolase, urease, acid phosphatase, and alkaline phosphatase responded positively to elevated CO2 concentrations. Generally, the enzyme activities were highly correlated with each other. Irrespective of the level of CO2, short-term negative influence of chlorpyriphos was observed on soil enzymes till day 7 of spray. Knowledge obtained from this study highlights that the elevated CO2 may negatively influence persistence of pesticide but will have positive effects on soil enzyme activities.

  13. Impact of land-use and long-term (>150 years) charcoal accumulation on microbial activity, biomass and community structure in temperate soils (Belgium).

    Science.gov (United States)

    Hardy, Brieuc; Cornelis, Jean-Thomas; Dufey, Joseph E.

    2015-04-01

    In the last decade, biochar has been increasingly investigated as a soil amendment for long-term soil carbon sequestration while improving soil fertility. On the short term, biochar application to soil generally increases soil respiration as well as microbial biomass and activity and affects significantly the microbial community structure. However, such effects are relatively short-term and tend to vanish over time. In our study, we investigated the long-term impact of charcoal accumulation and land-use on soil biota in temperate haplic Luvisols developed in the loess belt of Wallonia (Belgium). Charcoal-enriched soils were collected in the topsoil of pre-industrial (>150 years old) charcoal kilns in forest (4 sites) and cropland (5 sites). The topsoil of the adjacent charcoal-unaffected soils was sampled in a comparable way. Soils were characterized (pH, total, organic and inorganic C, total N, exchangeable Ca, Mg, K, Na, cation exchange capacity and available P) and natural soil organic matter (SOM) and black carbon (BC) contents were determined by differential scanning calorimetry. After rewetting at pF 2.5, soils were incubated during 140 days at 20 °C. At 70 days of incubation, 10 g of each soil were freeze dried in order to measure total microbial biomass and community structure by PLFA analysis. The PLFA dataset was analyzed by principal component analysis (PCA) while soil parameters were used as supplementary variables. For both agricultural and forest soils, the respiration rate is highly related to the total microbial biomass (R²=0.90). Both soil respiration and microbial biomass greatly depend on the SOM content, which indicates that the BC pool is relatively inert microbiologically. Land-use explains most of the variance in the PLFA dataset, largely governing the first principal component of the ACP. In forest soils, we observe a larger proportion of gram + bacteria, actinomycetes and an increased bacteria:fungi ratio compared to cropland, where gram

  14. Qualitative Analysis of Microbial Dynamics during Anaerobic Digestion of Microalgal Biomass in a UASB Reactor

    Directory of Open Access Journals (Sweden)

    Anna Doloman

    2017-01-01

    Full Text Available Anaerobic digestion (AD is a microbiologically coordinated process with dynamic relationships between bacterial players. Current understanding of dynamic changes in the bacterial composition during the AD process is incomplete. The objective of this research was to assess changes in bacterial community composition that coordinates with anaerobic codigestion of microalgal biomass cultivated on municipal wastewater. An upflow anaerobic sludge blanket reactor was used to achieve high rates of microalgae decomposition and biogas production. Samples of the sludge were collected throughout AD and extracted DNA was subjected to next-generation sequencing using methanogen mcrA gene specific and universal bacterial primers. Analysis of the data revealed that samples taken at different stages of AD had varying bacterial composition. A group consisting of Bacteroidales, Pseudomonadales, and Enterobacteriales was identified to be putatively responsible for the hydrolysis of microalgal biomass. The methanogenesis phase was dominated by Methanosarcina mazei. Results of observed changes in the composition of microbial communities during AD can be used as a road map to stimulate key bacterial species identified at each phase of AD to increase yield of biogas and rate of substrate decomposition. This research demonstrates a successful exploitation of methane production from microalgae without any biomass pretreatment.

  15. Qualitative Analysis of Microbial Dynamics during Anaerobic Digestion of Microalgal Biomass in a UASB Reactor

    Science.gov (United States)

    Doloman, Anna; Soboh, Yousef; Walters, Andrew J.; Sims, Ronald C.

    2017-01-01

    Anaerobic digestion (AD) is a microbiologically coordinated process with dynamic relationships between bacterial players. Current understanding of dynamic changes in the bacterial composition during the AD process is incomplete. The objective of this research was to assess changes in bacterial community composition that coordinates with anaerobic codigestion of microalgal biomass cultivated on municipal wastewater. An upflow anaerobic sludge blanket reactor was used to achieve high rates of microalgae decomposition and biogas production. Samples of the sludge were collected throughout AD and extracted DNA was subjected to next-generation sequencing using methanogen mcrA gene specific and universal bacterial primers. Analysis of the data revealed that samples taken at different stages of AD had varying bacterial composition. A group consisting of Bacteroidales, Pseudomonadales, and Enterobacteriales was identified to be putatively responsible for the hydrolysis of microalgal biomass. The methanogenesis phase was dominated by Methanosarcina mazei. Results of observed changes in the composition of microbial communities during AD can be used as a road map to stimulate key bacterial species identified at each phase of AD to increase yield of biogas and rate of substrate decomposition. This research demonstrates a successful exploitation of methane production from microalgae without any biomass pretreatment. PMID:29259629

  16. Rhizosphere soil microbial index of tree species in a coal mining ecosystem

    Energy Technology Data Exchange (ETDEWEB)

    Sinha, S.; Masto, R.E.; Ram, L.C.; Selvi, V.A.; Srivastava, N.K.; Tripathi, R.C.; George, J. [Central Institute of Mining & Fuel Research, Dhanbad (India)

    2009-09-15

    Microbial characterization of the tree rhizosphere provides important information relating to the screening of tree species for re-vegetation of degraded land. Rhizosphere soil samples collected from a few predominant tree species growing in the coal mining ecosystem of Dhanbad, India, were analyzed for soil organic carbon (SOC), mineralizable N, microbial biomass carbon (MBC), active microbial biomass carbon (AMBC), basal soil respiration (BSR), and soil enzyme activities (dehydrogenase, urease, catalase, phenol oxidase, and peroxidase). Principal component analysis was employed to derive a rhizosphere soil microbial index (RSMI) and accordingly, dehydrogenase, BSR/MBC, MBC/SOC, EC, phenol oxidase and AMBC were found to be the most critical properties. The observed values for the above properties were converted into a unitless score (0-1.00) and the scores were integrated into RSMI. The tree species could be arranged in decreasing order of the RSMI as: A. marmelos (0.718), A. indica (0.715), Bauhinia bauhinia (0.693), B. monosperma (0.611), E. jambolana (0.601), Moringa oleifera (0.565), Dalbergia sissoo (0.498), T indica (0.488), Morus alba (0.415), F religiosa (0.291), Eucalyptus sp. (0.232) and T grandis (0.181). It was concluded that tree species in coal mining areas had diverse effects on their respective rhizosphere microbial processes, which could directly or indirectly determine the survival and performance of the planted tree species in degraded coal mining areas. Tree species with higher RSMI values could be recommended for re-vegetation of degraded coal mining area.

  17. Biomass production from electricity using ammonia as an electron carrier in a reverse microbial fuel cell.

    Directory of Open Access Journals (Sweden)

    Wendell O Khunjar

    Full Text Available The storage of renewable electrical energy within chemical bonds of biofuels and other chemicals is a route to decreasing petroleum usage. A critical challenge is the efficient transfer of electrons into a biological host that can covert this energy into high energy organic compounds. In this paper, we describe an approach whereby biomass is grown using energy obtained from a soluble mediator that is regenerated electrochemically. The net result is a separate-stage reverse microbial fuel cell (rMFC that fixes CO₂ into biomass using electrical energy. We selected ammonia as a low cost, abundant, safe, and soluble redox mediator that facilitated energy transfer to biomass. Nitrosomonas europaea, a chemolithoautotroph, was used as the biocatalyst due to its inherent capability to utilize ammonia as its sole energy source for growth. An electrochemical reactor was designed for the regeneration of ammonia from nitrite, and current efficiencies of 100% were achieved. Calculations indicated that overall bioproduction efficiency could approach 2.7±0.2% under optimal electrolysis conditions. The application of chemolithoautotrophy for industrial bioproduction has been largely unexplored, and results suggest that this and related rMFC platforms may enable biofuel and related biochemical production.

  18. Microbial biomass and soil fauna during the decomposition of cover crops in no-tillage system

    Directory of Open Access Journals (Sweden)

    Luciano Colpo Gatiboni

    2011-08-01

    Full Text Available The decomposition of plant residues is a biological process mediated by soil fauna, but few studies have been done evaluating its dynamics in time during the process of disappearance of straw. This study was carried out in Chapecó, in southern Brazil, with the objective of monitoring modifications in soil fauna populations and the C content in the soil microbial biomass (C SMB during the decomposition of winter cover crop residues in a no-till system. The following treatments were tested: 1 Black oat straw (Avena strigosa Schreb.; 2 Rye straw (Secale cereale L.; 3 Common vetch straw (Vicia sativa L.. The cover crops were grown until full flowering and then cut mechanically with a rolling stalk chopper. The soil fauna and C content in soil microbial biomass (C SMB were assessed during the period of straw decomposition, from October 2006 to February 2007. To evaluate C SMB by the irradiation-extraction method, soil samples from the 0-10 cm layer were used, collected on eight dates, from before until 100 days after residue chopping. The soil fauna was collected with pitfall traps on seven dates up to 85 days after residue chopping. The phytomass decomposition of common vetch was faster than of black oat and rye residues. The C SMB decreased during the process of straw decomposition, fastest in the treatment with common vetch. In the common vetch treatment, the diversity of the soil fauna was reduced at the end of the decomposition process.

  19. Structural Studies of Biomass Degrading Enzyme Systems

    Energy Technology Data Exchange (ETDEWEB)

    Lunin, Vladimir V.; Alahuhta, Markus; Brunecky, Roman; Donohoe, Bryon; Xu, Qi; Bomble, Yannick J.; Himmel, Michael E.

    2014-08-05

    Renewable energy today comprises wind, photovoltaics, geothermal, and biofuels. Biomass is the leading source of renewable, sustainable energy used for the production of liquid transportation fuels. While the focus is shifting today from the ethanol towards next generation or advanced biofuels the real challenge however remains the same: reducing the recalcitrance of biomass to deconstruction, which yields the sugars needed for further processing.

  20. Nitrogen deposition and management practices increase soil microbial biomass carbon but decrease diversity in Moso bamboo plantations

    Science.gov (United States)

    Li, Quan; Song, Xinzhang; Gu, Honghao; Gao, Fei

    2016-06-01

    Because microbial communities play a key role in carbon (C) and nitrogen (N) cycling, changes in the soil microbial community may directly affect ecosystem functioning. However, the effects of N deposition and management practices on soil microbes are still poorly understood. We studied the effects of these two factors on soil microbial biomass carbon (MBC) and community composition in Moso bamboo plantations using high-throughput sequencing of the 16S rRNA gene. Plantations under conventional (CM) or intensive management (IM) were subjected to one of four N treatments for 30 months. IM and N addition, both separately and in combination, significantly increased soil MBC while decreasing bacterial diversity. However, increases in soil MBC were inhibited when N addition exceeded 60 kg N•ha-1•yr-1. IM increased the relative abundances of Actinobacteria and Crenarchaeota but decreased that of Acidobacteria. N addition increased the relative abundances of Acidobacteria, Crenarchaeota, and Actinobacteria but decreased that of Proteobacteria. Soil bacterial diversity was significantly related to soil pH, C/N ratio, and nitrogen and available phosphorus content. Management practices exerted a greater influence over regulation of the soil MBC and microbial diversity compared to that of N deposition in Moso bamboo plantations.

  1. Laccases as a Potential Tool for the Efficient Conversion of Lignocellulosic Biomass: A Review

    Directory of Open Access Journals (Sweden)

    Úrsula Fillat

    2017-05-01

    Full Text Available The continuous increase in the world energy and chemicals demand requires the development of sustainable alternatives to non-renewable sources of energy. Biomass facilities and biorefineries represent interesting options to gradually replace the present industry based on fossil fuels. Lignocellulose is the most promising feedstock to be used in biorefineries. From a sugar platform perspective, a wide range of fuels and chemicals can be obtained via microbial fermentation processes, being ethanol the most significant lignocellulose-derived fuel. Before fermentation, lignocellulose must be pretreated to overcome its inherent recalcitrant structure and obtain the fermentable sugars. Usually, harsh conditions are required for pretreatment of lignocellulose, producing biomass degradation and releasing different compounds that are inhibitors of the hydrolytic enzymes and fermenting microorganisms. Moreover, the lignin polymer that remains in pretreated materials also affects biomass conversion by limiting the enzymatic hydrolysis. The use of laccases has been considered as a very powerful tool for delignification and detoxification of pretreated lignocellulosic materials, boosting subsequent saccharification and fermentation processes. This review compiles the latest studies about the application of laccases as useful and environmentally friendly delignification and detoxification technology, highlighting the main challenges and possible ways to make possible the integration of these enzymes in future lignocellulose-based industries.

  2. Earthworm effects without earthworms: inoculation of raw organic matter with worm-worked substrates alters microbial community functioning.

    Science.gov (United States)

    Aira, Manuel; Domínguez, Jorge

    2011-01-27

    Earthworms are key organisms in organic matter decomposition because of the interactions they establish with soil microorganisms. They enhance decomposition rates through the joint action of direct effects (i.e. effects due to direct earthworm activity such as digestion, burrowing, etc) and indirect effects (i.e. effects derived from earthworm activities such as cast ageing). Here we test whether indirect earthworm effects affect microbial community functioning in the substrate, as when earthworms are present (i. e., direct effects). To address these questions we inoculated fresh organic matter (pig manure) with worm-worked substrates (vermicompost) produced by three different earthworm species. Two doses of each vermicompost were used (2.5 and 10%). We hypothesized that the presence of worm-worked material in the fresh organic matter will result in an inoculum of different microorganisms and nutrients. This inoculum should interact with microbial communities in fresh organic matter, thus promoting modifications similar to those found when earthworms are present. Inoculation of worm-worked substrates provoked significant increases in microbial biomass and enzyme activities (β-glucosidase, cellulase, phosphatase and protease). These indirect effects were similar to, although lower than, those obtained in pig manure with earthworms (direct and indirect earthworm effects). In general, the effects were not dose-dependent, suggesting the existence of a threshold at which they were triggered. Our data reveal that the relationships between earthworms and microorganisms are far from being understood, and suggest the existence of several positive feedbacks during earthworm activity as a result of the interactions between direct and indirect effects, since their combination produces stronger modifications to microbial biomass and enzyme activity.

  3. Earthworm effects without earthworms: inoculation of raw organic matter with worm-worked substrates alters microbial community functioning.

    Directory of Open Access Journals (Sweden)

    Manuel Aira

    Full Text Available BACKGROUND: Earthworms are key organisms in organic matter decomposition because of the interactions they establish with soil microorganisms. They enhance decomposition rates through the joint action of direct effects (i.e. effects due to direct earthworm activity such as digestion, burrowing, etc and indirect effects (i.e. effects derived from earthworm activities such as cast ageing. Here we test whether indirect earthworm effects affect microbial community functioning in the substrate, as when earthworms are present (i. e., direct effects. METHODOLOGY/PRINCIPAL FINDINGS: To address these questions we inoculated fresh organic matter (pig manure with worm-worked substrates (vermicompost produced by three different earthworm species. Two doses of each vermicompost were used (2.5 and 10%. We hypothesized that the presence of worm-worked material in the fresh organic matter will result in an inoculum of different microorganisms and nutrients. This inoculum should interact with microbial communities in fresh organic matter, thus promoting modifications similar to those found when earthworms are present. Inoculation of worm-worked substrates provoked significant increases in microbial biomass and enzyme activities (β-glucosidase, cellulase, phosphatase and protease. These indirect effects were similar to, although lower than, those obtained in pig manure with earthworms (direct and indirect earthworm effects. In general, the effects were not dose-dependent, suggesting the existence of a threshold at which they were triggered. CONCLUSION/SIGNIFICANCE: Our data reveal that the relationships between earthworms and microorganisms are far from being understood, and suggest the existence of several positive feedbacks during earthworm activity as a result of the interactions between direct and indirect effects, since their combination produces stronger modifications to microbial biomass and enzyme activity.

  4. Kinetic modeling of microbially-driven redox chemistry of radionuclides in subsurface environments: Coupling transport, microbial metabolism and geochemistry

    International Nuclear Information System (INIS)

    Wang, Yifeng; Papenguth, Hans W.

    2000-01-01

    Microbial degradation of organic matter is a driving force in many subsurface geochemical systems, and therefore may have significant impacts on the fate of radionuclides released into subsurface environments. In this paper, the authors present a general reaction-transport model for microbial metabolism, redox chemistry, and radionuclide migration in subsurface systems. The model explicitly accounts for biomass accumulation and the coupling of radionuclide redox reactions with major biogeochemical processes. Based on the consideration that the biomass accumulation in subsurface environments is likely to achieve a quasi-steady state, they have accordingly modified the traditional microbial growth kinetic equation. They justified the use of the biogeochemical models without the explicit representation of biomass accumulation, if the interest of modeling is in the net impact of microbial reactions on geochemical processes. They then applied their model to a scenario in which an oxic water flow containing both uranium and completing organic ligands is recharged into an oxic aquifer in a carbonate formation. The model simulation shows that uranium can be reduced and therefore immobilized in the anoxic zone created by microbial degradation

  5. Kinetic modeling of microbially-driven redox chemistry of radionuclides in subsurface environments: Coupling transport, microbial metabolism and geochemistry

    Energy Technology Data Exchange (ETDEWEB)

    WANG,YIFENG; PAPENGUTH,HANS W.

    2000-05-04

    Microbial degradation of organic matter is a driving force in many subsurface geochemical systems, and therefore may have significant impacts on the fate of radionuclides released into subsurface environments. In this paper, the authors present a general reaction-transport model for microbial metabolism, redox chemistry, and radionuclide migration in subsurface systems. The model explicitly accounts for biomass accumulation and the coupling of radionuclide redox reactions with major biogeochemical processes. Based on the consideration that the biomass accumulation in subsurface environments is likely to achieve a quasi-steady state, they have accordingly modified the traditional microbial growth kinetic equation. They justified the use of the biogeochemical models without the explicit representation of biomass accumulation, if the interest of modeling is in the net impact of microbial reactions on geochemical processes. They then applied their model to a scenario in which an oxic water flow containing both uranium and completing organic ligands is recharged into an oxic aquifer in a carbonate formation. The model simulation shows that uranium can be reduced and therefore immobilized in the anoxic zone created by microbial degradation.

  6. Vertical Integration of Biomass Saccharification of Enzymes for Sustainable Cellulosic Biofuel Production in a Biorefinery

    Energy Technology Data Exchange (ETDEWEB)

    Kumar, Manoj [DSM Innovation, Inc., San Francisco, CA (United States)

    2011-05-09

    These are a set of slides from this conference. Lignocellulosic biomass is the most abundant, least expensive renewable natural biological resource for the production of biobased products and bioenergy is important for the sustainable development of human civilization in 21st century. For making the fermentable sugars from lignocellulosic biomass, a reduction in cellulase production cost, an improvement in cellulase performance, and an increase in sugar yields are all vital to reduce the processing costs of biorefineries. Improvements in specific cellulase activities for non-complexed cellulase mixtures can be implemented through cellulase engineering based on rational design or directed evolution for each cellulase component enzyme, as well as on the reconstitution of cellulase components. In this paper, we will provide DSM's efforts in cellulase research and developments and focus on limitations. Cellulase improvement strategies based on directed evolution using screening on relevant substrates, screening for higher thermal tolerance based on activity screening approaches such as continuous culture using insoluble cellulosic substrates as a powerful selection tool for enriching beneficial cellulase mutants from the large library. We will illustrate why and how thermostable cellulases are vital for economic delivery of bioproducts from cellulosic biomass using biochemical conversion approach.

  7. Thermotolerant and mesophylic fungi from sugarcane bagasse and their prospection for biomass-degrading enzyme production

    Directory of Open Access Journals (Sweden)

    Bruna Silveira Lamanes dos Santos

    2015-09-01

    Full Text Available Nineteen fungi and seven yeast strains were isolated from sugarcane bagasse piles from an alcohol plant located at Brazilian Cerrado and identified up to species level on the basis of the gene sequencing of 5.8S-ITS and 26S ribosomal DNA regions. Four species were identified: Kluyveromyces marxianus, Aspergillus niger, Aspergillus sydowii and Aspergillus fumigatus, and the isolates were screened for the production of key enzymes in the saccharification of lignocellulosic material. Among them, three strains were selected as good producers of hemicellulolitic enzymes: A. niger (SBCM3, A. sydowii (SBCM7 and A. fumigatus (SBC4. The best β-xylosidase producer was A. niger SBCM3 strain. This crude enzyme presented optimal activity at pH 3.5 and 55 °C (141 U/g. For β-glucosidase and xylanase the best producer was A. fumigatus SBC4 strain, whose enzymes presented maximum activity at 60 °C and pH 3.5 (54 U/g and 4.0 (573 U/g, respectively. All these crude enzymes presented stability around pH 3.0–8.0 and up to 60 °C, which can be very useful in industrial processes that work at high temperatures and low pHs. These enzymes also exhibited moderate tolerance to ethanol and the sugars glucose and xylose. These similar characteristics among these fungal crude enzymes suggest that they can be used synergistically in cocktails in future studies of biomass conversion with potential application in several biotechnological sectors.

  8. Input related microbial carbon dynamic of soil organic matter in particle size fractions

    Science.gov (United States)

    Gude, A.; Kandeler, E.; Gleixner, G.

    2012-04-01

    This paper investigated the flow of carbon into different groups of soil microorganisms isolated from different particle size fractions. Two agricultural sites of contrasting organic matter input were compared. Both soils had been submitted to vegetation change from C3 (Rye/Wheat) to C4 (Maize) plants, 25 and 45 years ago. Soil carbon was separated into one fast-degrading particulate organic matter fraction (POM) and one slow-degrading organo-mineral fraction (OMF). The structure of the soil microbial community were investigated using phospholipid fatty acids (PLFA), and turnover of single PLFAs was calculated from the changes in their 13C content. Soil enzyme activities involved in the degradation of carbohydrates was determined using fluorogenic MUF (methyl-umbelliferryl phosphate) substrates. We found that fresh organic matter input drives soil organic matter dynamic. Higher annual input of fresh organic matter resulted in a higher amount of fungal biomass in the POM-fraction and shorter mean residence times. Fungal activity therefore seems essential for the decomposition and incorporation of organic matter input into the soil. As a consequence, limited litter input changed especially the fungal community favouring arbuscular mycorrhizal fungi. Altogether, supply and availability of fresh plant carbon changed the distribution of microbial biomass, the microbial community structure and enzyme activities and resulted in different priming of soil organic matter. Most interestingly we found that only at low input the OMF fraction had significantly higher calculated MRT for Gram-positive and Gram-negative bacteria suggesting high recycling of soil carbon or the use of other carbon sources. But on average all microbial groups had nearly similar carbon uptake rates in all fractions and both soils, which contrasted the turnover times of bulk carbon. Hereby the microbial carbon turnover was always faster than the soil organic carbon turnover and higher carbon input

  9. Restoration of areas degraded by alluvial sand mining: use of soil microbiological activity and plant biomass growth to assess evolution of restored riparian vegetation.

    Science.gov (United States)

    Venson, Graziela R; Marenzi, Rosemeri C; Almeida, Tito César M; Deschamps-Schmidt, Alexandre; Testolin, Renan C; Rörig, Leonardo R; Radetski, Claudemir M

    2017-03-01

    River or alluvial sand mining is causing a variety of environmental problems in the Itajaí-açú river basin in Santa Catarina State (south of Brazil). When this type of commercial activity degrades areas around rivers, environmental restoration programs need to be executed. In this context, the aim of this study was to assess the evolution of a restored riparian forest based on data on the soil microbial activity and plant biomass growth. A reference site and three sites with soil degradation were studied over a 3-year period. Five campaigns were performed to determine the hydrolysis of the soil enzyme fluorescein diacetate (FDA), and the biomass productivity was determined at the end of the studied period. The variation in the enzyme activity for the different campaigns at each site was low, but this parameter did differ significantly according to the site. Well-managed sites showed the highest biomass productivity, and this, in turn, showed a strong positive correlation with soil enzyme activity. In conclusion, soil enzyme activity could form the basis for monitoring and the early prediction of the success of vegetal restoration programs, since responses at the higher level of biological organization take longer, inhibiting the assessment of the project within an acceptable time frame.

  10. Microbial diversity and digestive enzyme activities in the gut of earthworms found in sawmill industries in Abeokuta, Nigeria

    Directory of Open Access Journals (Sweden)

    Bamidele Julius A.

    2014-09-01

    Full Text Available The growing demand for wood has resulted in large volumes of wood wastes that are daily released to the soil from the activities of sawmills in South-Western Nigeria. In an attempt to setup a bioremediation model for sawdust, this study therefore aimed at evaluating microbial diversity, and the level of digestive enzymes in the gut of earthworms (Eudrilus eugeniae, Libyodrilus violaceous and Hyperiodrilus africanus of sawmill origin. Four major sawmills located in Abeokuta (7o9’12” N - 3o19’35” E, namely Lafenwa, Sapon, Isale-Ake and Kotopo sawmills were used for this study. The arboretum of the Federal University of Agriculture, Abeokuta was used as control. Gut microbial analysis was carried out using the pour-plate method while digestive enzyme activities in the earthworm guts were done by the spectrophotometric method. Higher microbial counts (28.5±0.1x10³-97.0±0.1x10³cfu for bacteria and 7.0±0.1x10³-96.0±0.1x10³cfu for fungi and microbial diversity were recorded in the gut of earthworms of the sawmill locations than those of the control site (17.5±0.1x10³cfu for bacteria and 4.5±0.1x10³cfu for fungi. Streptococcus mutans and Proteus spp.were common in the gut of E. eugeniae, and L. violaceous from the study sawmills, while Streptococcus mutans were also identified in H. africanus, but absent in the gut of E. eugeniae from the control site. Cellulase (48.67±0.02mg/g and lipase (1.81±0.01mg/g activities were significantly higher (p<0.05 in the gut of earthworms from the control site than those of the study sawmills. Furthermore, amylase (α and β activity was highest in the gut of earthworms from the sawmills. Variations observed in the gut microbial and digestive enzyme activities of earthworms from the study sawmills as compared to the control site suggests that earthworms, especially E. eugeniae, could be a better organism for use as bioremediator of wood wastes. Rev. Biol. Trop. 62 (3: 1241-1249. Epub 2014 September

  11. Microbial diversity and digestive enzyme activities in the gut of earthworms found in sawmill industries in Abeokuta, Nigeria.

    Science.gov (United States)

    Bamidele, Julius A; Idowu, Adewunmi B; Ademolu, Kehinde O; Atayese, Adijat O

    2014-09-01

    The growing demand for wood has resulted in large volumes of wood wastes that are daily released to the soil from the activities of sawmills in South-Western Nigeria. In an attempt to setup a bioremediation model for sawdust, this study therefore aimed at evaluating microbial diversity, and the level of digestive enzymes in the gut of earthworms (Eudrilus eugeniae, Libyodrilus violaceous and Hyperiodrilus africanus) of sawmill origin. Four major sawmills located in Abeokuta (7°9'12" N- 3°19'35" E), namely Lafenwa, Sapon, Isale-Ake and Kotopo sawmills were used for this study. The arboretum of the Federal University of Agriculture, Abeokuta was used as control. Gut microbial analysis was carried out using the pour-plate method while digestive enzyme activities in the earthworm guts were done by the spectrophotometric method. Higher microbial counts (28.5 ± 0.1 x 10(3)-97.0 ± 0.1 x 10(3) cfu for bacteria and 7.0 ± 0.1x 10(3)-96.0 ± 0.1 x 10(3) cfu for fungi) and microbial diversity were recorded in the gut of earthworms of the sawmill locations than those of the control site (17.5 ± 0.1 x10(3) cfu for bacteria and 4.5 ± 0.1 x 10(3) cfu for fungi). Streptococcus mutans and Proteus spp. were common in the gut of E. eugeniae, and L. violaceous from the study sawmills, while Streptococcus mutans were also identified in H. africanus, but absent in the gut of E. eugeniae from the control site. Cellulase (48.67 ± 0.02 mg/g) and lipase (1.81 ± 0.01 mg/g) activities were significantly higher (p earthworms from the control site than those of the study sawmills. Furthermore, amylase (α and β) activity was highest in the gut of earthworms from the sawmills. Variations observed in the gut microbial and digestive enzyme activities of earthworms from the study sawmills as compared to the control site suggests that earthworms, especially E. eugeniae, could be a better organism for use as bioremediator of wood wastes.

  12. Effects of microcystins contamination on soil enzyme activities and microbial community in two typical lakeside soils.

    Science.gov (United States)

    Cao, Qing; Steinman, Alan D; Su, Xiaomei; Xie, Liqiang

    2017-12-01

    A 30-day indoor incubation experiment was conducted to investigate the effects of different concentrations of microcystin (1, 10, 100 and 1000 μg eq. MC-LR L -1 ) on soil enzyme activity, soil respiration, physiological profiles, potential nitrification, and microbial abundance (total bacteria, total fungi, ammonia-oxidizing bacteria and archaea) in two lakeside soils in China (Soil A from the lakeside of Lake Poyanghu at Jiujiang; Soil B from the lakeside of Lake Taihu at Suzhou). Of the enzymes tested, only phenol oxidase activity was negatively affected by microcystin application. In contrast, dehydrogenase activity was stimulated in the 1000 μg treatment, and a stimulatory effect also occurred with soil respiration in contaminated soil. The metabolic profiles of the microbial communities indicated that overall carbon metabolic activity in the soils treated with high microcystin concentrations was inhibited, and high concentrations of microcystin also led to different patterns of potential carbon utilization. High microcystin concentrations (100, 1000 μg eq. MC-LR L -1 in Soil A; 10, 100 1000 μg eq. MC-LR L -1 in Soil B) significantly decreased soil potential nitrification rate. Furthermore, the decrease in soil potential nitrification rate was positively correlated with the decrease of the amoA gene abundance, which corresponds to the ammonia-oxidizing bacterial community. We conclude that application of microcystin-enriched irrigation water can significantly impact soil microbial community structure and function. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Differential sensitivity of total and active soil microbial communities to drought and forest management.

    Science.gov (United States)

    Bastida, Felipe; Torres, Irene F; Andrés-Abellán, Manuela; Baldrian, Petr; López-Mondéjar, Rubén; Větrovský, Tomáš; Richnow, Hans H; Starke, Robert; Ondoño, Sara; García, Carlos; López-Serrano, Francisco R; Jehmlich, Nico

    2017-10-01

    Climate change will affect semiarid ecosystems through severe droughts that increase the competition for resources in plant and microbial communities. In these habitats, adaptations to climate change may consist of thinning-that reduces competition for resources through a decrease in tree density and the promotion of plant survival. We deciphered the functional and phylogenetic responses of the microbial community to 6 years of drought induced by rainfall exclusion and how forest management affects its resistance to drought, in a semiarid forest ecosystem dominated by Pinus halepensis Mill. A multiOMIC approach was applied to reveal novel, community-based strategies in the face of climate change. The diversity and the composition of the total and active soil microbiome were evaluated by 16S rRNA gene (bacteria) and ITS (fungal) sequencing, and by metaproteomics. The microbial biomass was analyzed by phospholipid fatty acids (PLFAs), and the microbially mediated ecosystem multifunctionality was studied by the integration of soil enzyme activities related to the cycles of C, N, and P. The microbial biomass and ecosystem multifunctionality decreased in drought-plots, as a consequence of the lower soil moisture and poorer plant development, but this decrease was more notable in unthinned plots. The structure and diversity of the total bacterial community was unaffected by drought at phylum and order level, but did so at genus level, and was influenced by seasonality. However, the total fungal community and the active microbial community were more sensitive to drought and were related to ecosystem multifunctionality. Thinning in plots without drought increased the active diversity while the total diversity was not affected. Thinning promoted the resistance of ecosystem multifunctionality to drought through changes in the active microbial community. The integration of total and active microbiome analyses avoids misinterpretations of the links between the soil microbial

  14. Leaf-Cutter Ant Fungus Gardens Are Biphasic Mixed Microbial Bioreactors That Convert Plant Biomass to Polyols with Biotechnological Applications

    Science.gov (United States)

    Somera, Alexandre F.; Lima, Adriel M.; dos Santos-Neto, Álvaro J.; Lanças, Fernando M.

    2015-01-01

    Leaf-cutter ants use plant matter to culture the obligate mutualistic basidiomycete Leucoagaricus gongylophorus. This fungus mediates ant nutrition on plant resources. Furthermore, other microbes living in the fungus garden might also contribute to plant digestion. The fungus garden comprises a young sector with recently incorporated leaf fragments and an old sector with partially digested plant matter. Here, we show that the young and old sectors of the grass-cutter Atta bisphaerica fungus garden operate as a biphasic solid-state mixed fermenting system. An initial plant digestion phase occurred in the young sector in the fungus garden periphery, with prevailing hemicellulose and starch degradation into arabinose, mannose, xylose, and glucose. These products support fast microbial growth but were mostly converted into four polyols. Three polyols, mannitol, arabitol, and inositol, were secreted by L. gongylophorus, and a fourth polyol, sorbitol, was likely secreted by another, unidentified, microbe. A second plant digestion phase occurred in the old sector, located in the fungus garden core, comprising stocks of microbial biomass growing slowly on monosaccharides and polyols. This biphasic operation was efficient in mediating symbiotic nutrition on plant matter: the microbes, accounting for 4% of the fungus garden biomass, converted plant matter biomass into monosaccharides and polyols, which were completely consumed by the resident ants and microbes. However, when consumption was inhibited through laboratory manipulation, most of the plant polysaccharides were degraded, products rapidly accumulated, and yields could be preferentially switched between polyols and monosaccharides. This feature might be useful in biotechnology. PMID:25911490

  15. Feedbacks Between Soil Structure and Microbial Activities in Soil

    Science.gov (United States)

    Bailey, V. L.; Smith, A. P.; Fansler, S.; Varga, T.; Kemner, K. M.; McCue, L. A.

    2017-12-01

    Soil structure provides the physical framework for soil microbial habitats. The connectivity and size distribution of soil pores controls the microbial access to nutrient resources for growth and metabolism. Thus, a crucial component of soil research is how a soil's three-dimensional structure and organization influences its biological potential on a multitude of spatial and temporal scales. In an effort to understand microbial processes at scale more consistent with a microbial community, we have used soil aggregates as discrete units of soil microbial habitats. Our research has shown that mean pore diameter (x-ray computed tomography) of soil aggregates varies with the aggregate diameter itself. Analyzing both the bacterial composition (16S) and enzyme activities of individual aggregates showed significant differences in the relative abundances of key members the microbial communities associated with high enzyme activities compared to those with low activities, even though we observed no differences in the size of the biomass, nor in the overall richness or diversity of these communities. We hypothesize that resources and substrates have stimulated key populations in the aggregates identified as highly active, and as such, we conducted further research that explored how such key populations (i.e. fungal or bacterial dominated populations) alter pathways of C accumulation in aggregate size domains and microbial C utilization. Fungi support and stabilize soil structure through both physical and chemical effects of their hyphal networks. In contrast, bacterial-dominated communities are purported to facilitate micro- and fine aggregate stabilization. Here we quantify the direct effects fungal versus bacterial dominated communities on aggregate formation (both the rate of aggregation and the quality, quantity and distribution of SOC contained within aggregates). A quantitative understanding of the different mechanisms through which fungi or bacteria shape aggregate

  16. Seasonal variations and effects of nutrient applications on N and P and microbial biomass under two temperate heathland plants

    DEFF Research Database (Denmark)

    Nielsen, Pia Lund; Andresen, Louise Christoffersen; Michelsen, Anders

    2009-01-01

    . The microbial biomass on the other hand was positively related to soil water content in fertilized plots indicating that this was due to an indirect effect of enhanced nutrient availability. Microbial N and P pools were respectively 1000 and 100 times higher than the pool of inorganic N and P, and microbes...... this process. In this study the soil properties under two dominant heathland plants, the dwarf shrub Calluna vulgaris and the grass Deschampsia flexuosa, were investigated, with focus on nutrient content in the organic top soil and soil microbes during the main growing season and effects of nutrient amendments...... therefore may play an important role in regulating plant nutrient supply. Judged from responses of inorganic and microbial N and P concentrations to added N and P, N seemed to limit C. vulgaris and soil microbes below while P seemed to limit D. flexuosa and soil microbes below this species. There were lower...

  17. Changes in microbial populations and enzyme activities during the bioremediation of oil-contaminated soil.

    Science.gov (United States)

    Lin, Xin; Li, Xiaojun; Sun, Tieheng; Li, Peijun; Zhou, Qixing; Sun, Lina; Hu, Xiaojun

    2009-10-01

    In the process of bioremediation in the soil contaminated by different oil concentrations, the changes in the microbial numbers (bacteria and fungi) and the enzyme (catalase (CAT), polyphenol oxidase (PPO) and lipase) activities were evaluated over a 2-year period. The results showed that the microbial numbers after 2-year bioremediation were one to ten times higher than those in the initial. The changes in the bacterial and the fungal populations were different during the bioremediation, and the highest microbial numbers for bacteria and fungi were 5.51 x 10(9) CFU g(-1) dry soil in treatment 3 (10,000 mg kg(-1)) in the initial and 5.54 x 10(5) CFU g(-1) dry soil in treatment 5 (50,000 mg kg(-1)) after the 2-year bioremediation period, respectively. The CAT and PPO activities in the contaminated soil decreased with increasing oil concentration, while the lipase activity increased. The activities of CAT and PPO improved after the bioremediation, but lipase activity was on the contrary. The CAT activity was more sensible to the oil than others, and could be alternative to monitor the bioremediation process.

  18. Enhancing Cellulase Commercial Performance for the Lignocellulosic Biomass Industry

    Energy Technology Data Exchange (ETDEWEB)

    Jarnigan, Alisha [Danisco, US Inc., Copenhagen (Denmark)

    2016-06-07

    Cellulase enzyme loading (Bt-G) for the economic conversion of lignocellulosic biomass to ethanol is on of the key challenges identified in the Biomass Program of DOE/EERE. The goal of Danisco’s project which ran from 2008 to 2012, was to address the technical challenge by creating more efficient enzyme that could be used at lower doses, thus reducing the enzymes’ cost contribution to the conversio process. We took the approach of protein engineering of cellulase enzymes to overcome the enzymati limitations in the system of cellulosic-hydrolyzing enzymes to improve performance in conversion o biomass, thereby creating a more effective enzyme mix.

  19. A Model of the Effect of the Microbial Biomass on the Isotherm of the Fermenting Solids in Solid-State Fermentation

    Directory of Open Access Journals (Sweden)

    Barbara Celuppi Marques

    2006-01-01

    Full Text Available We compare isotherms for soybeans and soybeans fermented with Rhizopus oryzae, showing that in solid-state fermentation the biomass affects the isotherm of the fermenting solids. Equations are developed to calculate, for a given overall water content of the fermenting solids, the water contents of the biomass and residual substrate, as well as the water activity. A case study, undertaken using a mathematical model of a well-mixed bioreactor, shows that if water additions are made on the basis of the assumption that fermenting solids have the same isotherm as the substrate itself, poor growth can result since the added water does not maintain the water activity at levels favorable for growth. We conclude that the effect of the microbial biomass on the isotherm of the fermenting solids must be taken into account in mathematical models of solid-state fermentation bioreactors.

  20. Microbial electrolysis cells as innovative technology for hydrogen production

    International Nuclear Information System (INIS)

    Chorbadzhiyska, Elitsa; Hristov, Georgi; Mitov, Mario; Hubenova, Yolina

    2011-01-01

    Hydrogen production is becoming increasingly important in view of using hydrogen in fuel cells. However, most of the production of hydrogen so far comes from the combustion of fossil fuels and water electrolysis. Microbial Electrolysis Cell (MEC), also known as Bioelectrochemically Assisted Microbial Reactor, is an ecologically clean, renewable and innovative technology for hydrogen production. Microbial electrolysis cells produce hydrogen mainly from waste biomass assisted by various bacteria strains. The principle of MECs and their constructional elements are reviewed and discussed. Keywords: microbial Electrolysis Cells, hydrogen production, waste biomass purification

  1. Novel enzymes for the degradation of cellulose

    Directory of Open Access Journals (Sweden)

    Horn Svein

    2012-07-01

    Full Text Available Abstract The bulk terrestrial biomass resource in a future bio-economy will be lignocellulosic biomass, which is recalcitrant and challenging to process. Enzymatic conversion of polysaccharides in the lignocellulosic biomass will be a key technology in future biorefineries and this technology is currently the subject of intensive research. We describe recent developments in enzyme technology for conversion of cellulose, the most abundant, homogeneous and recalcitrant polysaccharide in lignocellulosic biomass. In particular, we focus on a recently discovered new type of enzymes currently classified as CBM33 and GH61 that catalyze oxidative cleavage of polysaccharides. These enzymes promote the efficiency of classical hydrolytic enzymes (cellulases by acting on the surfaces of the insoluble substrate, where they introduce chain breaks in the polysaccharide chains, without the need of first “extracting” these chains from their crystalline matrix.

  2. [Effects of litter and root exclusion on soil microbial community composition and function of four plantations in subtropical sandy coastal plain area, China].

    Science.gov (United States)

    Sang, Chang Peng; Wan, Xiao Hua; Yu, Zai Peng; Wang, Min Huang; Lin, Yu; Huang, Zhi Qun

    2017-04-18

    We conducted detritus input and removal treatment (DIRT) to examine the effects of shifting above- and belowground carbon (C) inputs on soil microbial biomass, community composition and function in subtropical Pinus elliottii, Eucalyptus urophylla × Eucalyptus grandis, Acacia aulacocarpa and Casuarina equisetifolia coastal sandy plain forests, and the treatments included: root trenching, litter removal and control. Up to September 2015, one year after the experiment began, we collected the 0-10 cm soil samples from each plot. Phospholipid fatty acid (PLFA) analysis was used to characterize the microbial community composition, and micro-hole enzymatic detection technology was utilized to determine the activity of six kinds of soil enzymes. Results showed that changes in microbial biomass induced by the C input manipulations differed among tree species, and mainly affected by litter and root qualily. In E. urophylla × E. grandis stands, root trenching significantly decreased the contents of total PLFAs, Gram-positive bacteria, Gram-negative bacteria, fungi and actinomycetes by 31%, 30%, 32%, 36% and 26%, respectively. Litter removal reduced the contents of Gram-positive bacteria, fungi and actinomycetes by 24%, 27% and 24%, respectively. However, C input manipulations had no significant effect on soil microbial biomassunder other three plantations. According to the effect of C input manipulations on soil microbial community structure, litter and root exclusion decreased fungi abundance and increased actinomycetes abundance. Different treatments under different plantations resulted in various soil enzyme activities. Litter removal significantly decreased the activities of cellobiohydrolase, β-glucosidase, acid phosphatase and N-acetyl-β-d-glucosaminidase of P. elliottii, A. aulacocarpa and C. equisetifolia, root exclusion only decreased and increased the activities of β-glucosidase in P. elliottii and A. aulacocarpa forest soils, respectively. Litter removal also

  3. Root carbon decomposition and microbial biomass response at different soil depths

    Science.gov (United States)

    Rumpel, C.

    2012-12-01

    The relationship between root litter addition and soil organic matter (SOM) formation in top- versus subsoils is unknown. The aim of this study was to investigate root litter decomposition and stabilisation in relation to microbial parameters in different soil depths. Our conceptual approach included incubation of 13C-labelled wheat roots at 30, 60 and 90 cm soil depth for 36 months under field conditions. Quantitative root carbon contribution to SOM was assessed, changes of bulk root chemistry studied by solid-state 13C NMR spectroscopy and lignin content and composition was assessed after CuO oxidation. Compound-specific isotope analysis allowed to assess the role of root lignin for soil C storage in the different soil depths. Microbial biomass and community structure was determined after DNA extraction. After three years of incubation, O-alkyl C most likely assigned to polysaccharides decreased in all soil depth compared to the initial root material. The degree of root litter decomposition assessed by the alkyl/O-alkyl ratio decreased with increasing soil depth, while aryl/O-alkyl ratio was highest at 60 cm depth. Root-derived lignin showed depth specific concentrations (30 fungi contribution increased after root litter addition. Their community structure changed after root litter addition and showed horizon specific dynamics. Our study shows that root litter addition can contribute to C storage in subsoils but did not influence C storage in topsoil. We conclude that specific conditions of single soil horizons have to be taken into account if root C dynamics are to be fully understood.

  4. Combinational effects of sulfomethoxazole and copper on soil microbial community and function.

    Science.gov (United States)

    Liu, Aiju; Cao, Huansheng; Yang, Yan; Ma, Xiaoxuan; Liu, Xiao

    2016-03-01

    Sulfonamides and Cu are largely used feed additives in poultry farm. Subsequently, they are spread onto agricultural soils together with contaminated manure used as fertilizer. Both sulfonamides and Cu affect the soil microbial community. However, an interactive effect of sulfonamides and Cu on soil microorganisms is not well understood. Therefore, a short-time microcosm experiment was conducted to investigate the interaction of veterinary antibiotic sulfomethoxazole (SMX) and Cu on soil microbial structure composition and functions. To this end, selected concentrations of SMX (0, 5, and 50 mg kg(-1)) and Cu (0, 300, and 500 mg kg(-1)) were combined, respectively. Clear dose-dependent effects of SMX on microbial biomass and basal respiration were determined, and these effects were amplified in the presence of additional Cu. For activities of soil enzymes including β-glucosidase, urease, and protease, clear reducing effects were determined in soil samples containing 5 or 50 mg kg(-1) of SMX, and the interaction of SMX and Cu was significant, particularly in soil samples containing 50 mg kg(-1) SMX or 500 mg kg(-1) Cu. SMX amendments, particularly in combination with Cu, significantly reduced amounts of the total, bacterial, and fungal phospholipid fatty acids (PLFAs) in soil. Moreover, the derived ratio of bacteria to fungi decreased significantly with incremental SMX and Cu, and principal component analysis of the PLFAs showed that soil microbial composition was significantly affected by SMX interacted with Cu at 500 mg kg(-1). All of these results indicated that the interaction of SMX and Cu was synergistic to amplify the negative effect of SMX on soil microbial biomass, structural composition, and even the enzymatic function.

  5. Manipulating soil microbial communities in extensive green roof substrates.

    Science.gov (United States)

    Molineux, Chloe J; Connop, Stuart P; Gange, Alan C

    2014-09-15

    There has been very little investigation into the soil microbial community on green roofs, yet this below ground habitat is vital for ecosystem functioning. Green roofs are often harsh environments that would greatly benefit from having a healthy microbial system, allowing efficient nutrient cycling and a degree of drought tolerance in dry summer months. To test if green roof microbial communities could be manipulated, we added mycorrhizal fungi and a microbial mixture ('compost tea') to green roof rootzones, composed mainly of crushed brick or crushed concrete. The study revealed that growing media type and depth play a vital role in the microbial ecology of green roofs. There are complex relationships between depth and type of substrate and the biomass of different microbial groups, with no clear pattern being observed. Following the addition of inoculants, bacterial groups tended to increase in biomass in shallower substrates, whereas fungal biomass change was dependent on depth and type of substrate. Increased fungal biomass was found in shallow plots containing more crushed concrete and deeper plots containing more crushed brick where compost tea (a live mixture of beneficial bacteria) was added, perhaps due to the presence of helper bacteria for arbuscular mycorrhizal fungi (AMF). Often there was not an additive affect of the microbial inoculations but instead an antagonistic interaction between the added AM fungi and the compost tea. This suggests that some species of microbes may not be compatible with others, as competition for limited resources occurs within the various substrates. The overall results suggest that microbial inoculations of green roof habitats are sustainable. They need only be done once for increased biomass to be found in subsequent years, indicating that this is a novel and viable method of enhancing roof community composition. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Screening and Molecular Identification of New Microbial Strains for Production of Enzymes of Biotechnological Interest

    Directory of Open Access Journals (Sweden)

    Imen Ghazala

    Full Text Available ABSTRACT: This research focused on isolation, identification and characterization of new strains of fungi and bacteria, which were able to produce extracellular xylanase, mannanase, pectinase and α-amylase. Fungi isolates were identified on the basis of analyses of 18S gene sequencing and internal transcribed spacer region. The closest phylogenetic neighbors according to 18S gene sequence and ITS region data for the two isolates M1 and SE were Aspergillus fumigatus and Aspergillus sydowii, respectively. I4 was identified as Bacillus mojavensis on the basis of the 16S rRNA gene sequencing and biochemical properties. The enzyme production was evaluated by cultivating the isolated microorganisms in liquid-state bioprocess using wheat bran as carbon source. Two fungi (M1, and SE and one bacterium (I4 strains were found to be xylanase producer, and several were proven to be outstanding producers of microbial xylanase. The strains producing xylanase secreted variable amounts of starch-debranching enzymes and produced low level β-mannan-degrading enzyme systems. The bacterium strain was found to be capable of producing pectinolytic enzymes on wheat bran at high level. Some of the strains have good potential for use as sources of important industrial enzymes.

  7. Understanding and regulation of microbial lignolysis for renewable platform chemicals

    Energy Technology Data Exchange (ETDEWEB)

    Singh, Seema [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Hudson, Corey [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Turner, Kevin. [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Tran-Gyamfi, Mary Bao [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Williams, Kelly Porter [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Powell, Amy Jo [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Alam, Todd Michael [Sandia National Lab. (SNL-CA), Livermore, CA (United States)

    2014-01-01

    Lignin is often overlooked in the valorization of lignocellulosic biomass, but lignin-based materials and chemicals represent potential value-added products for biorefineries that could significantly improve the economics of a biorefinery. Fluctuating crude oil prices and changing fuel specifications are some of the driving factors to develop new technologies that could be used to convert polymeric lignin into low molecular weight lignin and or monomeric aromatic feedstocks to assist in the displacement of the current products associated with the conversion of a whole barrel of oil. Our project of understanding microbial lignolysis for renewable platform chemicals aimed to understand microbial and enzymatic lignolysis processes to break down lignin for conversion into commercially viable drop-in fuels. We developed novel lignin analytics to interrogate enzymatic and microbial lignolysis of native polymeric lignin and established a detailed understanding of lignolysis as a function of fungal enzyme, microbes and endophytes. Bioinformatics pipeline was developed for metatranscryptomic analysis of aridland ecosystem for investigating the potential discovery of new lignolysis gene and gene products.

  8. MICROBIAL CHARACTERISTICS OF SOILS UNDER AN INTEGRATED CROP-LIVESTOCK SYSTEM

    Directory of Open Access Journals (Sweden)

    Andréa Scaramal da Silva

    2015-02-01

    Full Text Available Integrated crop-livestock systems (ICLs are a viable strategy for the recovery and maintenance of soil characteristics. In the present study, an ICL experiment was conducted by the Instituto Agronômico do Paraná in the municipality of Xambre, Parana (PR, Brazil, to evaluate the effects of various grazing intensities. The objective of the present study was to quantify the levels of microbial biomass carbon (MBC and soil enzymatic activity in an ICL of soybean (summer and Brachiaria ruziziensis (winter, with B. ruziziensis subjected to various grazing intensities. Treatments consisted of varying pasture heights and grazing intensities (GI: 10, 20, 30, and 40 cm (GI-10, GI-20, GI-30, and GI-40, respectively and a no grazing (NG control. The microbial characteristics analysed were MBC, microbial respiration (MR, metabolic quotient (qCO2, the activities of acid phosphatase, β-glucosidase, arylsuphatase, and cellulase, and fluorescein diacetate (FDA hydrolysis. Following the second grazing cycle, the GI-20 treatment (20-cm - moderate grazing intensity contained the highest MBC concentrations and lowest qCO2 concentrations. Following the second soybean cycle, the treatment with the highest grazing intensity (GI-10 contained the lowest MBC concentration. Soil MBC concentrations in the pasture were favoured by the introduction of animals to the system. High grazing intensity (10-cm pasture height during the pasture cycle may cause a decrease in soil MBC and have a negative effect on the microbial biomass during the succeeding crop. Of all the enzymes analyzed, only arylsuphatase and cellulase activities were altered by ICL management, with differences between the moderate grazing intensity (GI-20 and no grazing (NG treatments.

  9. Controlled expression of pectic enzymes in Arabidopsis thaliana enhances biomass conversion without adverse effects on growth.

    Science.gov (United States)

    Tomassetti, Susanna; Pontiggia, Daniela; Verrascina, Ilaria; Reca, Ida Barbara; Francocci, Fedra; Salvi, Gianni; Cervone, Felice; Ferrari, Simone

    2015-04-01

    Lignocellulosic biomass from agriculture wastes is a potential source of biofuel, but its use is currently limited by the recalcitrance of the plant cell wall to enzymatic digestion. Modification of the wall structural components can be a viable strategy to overcome this bottleneck. We have previously shown that the expression of a fungal polygalacturonase (pga2 from Aspergillus niger) in Arabidopsis and tobacco plants reduces the levels of de-esterified homogalacturonan in the cell wall and significantly increases saccharification efficiency. However, plants expressing pga2 show stunted growth and reduced biomass production, likely as a consequence of an extensive loss of pectin integrity during the whole plant life cycle. We report here that the expression in Arabidopsis of another pectic enzyme, the pectate lyase 1 (PL1) of Pectobacterium carotovorum, under the control of a chemically inducible promoter, results, after induction of the transgene, in a saccharification efficiency similar to that of plants expressing pga2. However, lines with high levels of transgene induction show reduced growth even in the absence of the inducer. To overcome the problem of plant fitness, we have generated Arabidopsis plants that express pga2 under the control of the promoter of SAG12, a gene expressed only during senescence. These plants expressed pga2 only at late stages of development, and their growth was comparable to that of WT plants. Notably, leaves and stems of transgenic plants were more easily digested by cellulase, compared to WT plants, only during senescence. Expression of cell wall-degrading enzymes at the end of the plant life cycle may be therefore a useful strategy to engineer crops unimpaired in biomass yield but improved for bioconversion. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. Advances in High Throughput Screening of Biomass Recalcitrance (Poster)

    Energy Technology Data Exchange (ETDEWEB)

    Turner, G. B.; Decker, S. R.; Tucker, M. P.; Law, C.; Doeppke, C.; Sykes, R. W.; Davis, M. F.; Ziebell, A.

    2012-06-01

    This was a poster displayed at the Symposium. Advances on previous high throughput screening of biomass recalcitrance methods have resulted in improved conversion and replicate precision. Changes in plate reactor metallurgy, improved preparation of control biomass, species-specific pretreatment conditions, and enzymatic hydrolysis parameters have reduced overall coefficients of variation to an average of 6% for sample replicates. These method changes have improved plate-to-plate variation of control biomass recalcitrance and improved confidence in sugar release differences between samples. With smaller errors plant researchers can have a higher degree of assurance more low recalcitrance candidates can be identified. Significant changes in plate reactor, control biomass preparation, pretreatment conditions and enzyme have significantly reduced sample and control replicate variability. Reactor plate metallurgy significantly impacts sugar release aluminum leaching into reaction during pretreatment degrades sugars and inhibits enzyme activity. Removal of starch and extractives significantly decreases control biomass variability. New enzyme formulations give more consistent and higher conversion levels, however required re-optimization for switchgrass. Pretreatment time and temperature (severity) should be adjusted to specific biomass types i.e. woody vs. herbaceous. Desalting of enzyme preps to remove low molecular weight stabilizers and improved conversion levels likely due to water activity impacts on enzyme structure and substrate interactions not attempted here due to need to continually desalt and validate precise enzyme concentration and activity.

  11. Direct conversion of plant biomass to ethanol by engineered Caldicellulosiruptor bescii.

    Science.gov (United States)

    Chung, Daehwan; Cha, Minseok; Guss, Adam M; Westpheling, Janet

    2014-06-17

    Ethanol is the most widely used renewable transportation biofuel in the United States, with the production of 13.3 billion gallons in 2012 [John UM (2013) Contribution of the Ethanol Industry to the Economy of the United States]. Despite considerable effort to produce fuels from lignocellulosic biomass, chemical pretreatment and the addition of saccharolytic enzymes before microbial bioconversion remain economic barriers to industrial deployment [Lynd LR, et al. (2008) Nat Biotechnol 26(2):169-172]. We began with the thermophilic, anaerobic, cellulolytic bacterium Caldicellulosiruptor bescii, which efficiently uses unpretreated biomass, and engineered it to produce ethanol. Here we report the direct conversion of switchgrass, a nonfood, renewable feedstock, to ethanol without conventional pretreatment of the biomass. This process was accomplished by deletion of lactate dehydrogenase and heterologous expression of a Clostridium thermocellum bifunctional acetaldehyde/alcohol dehydrogenase. Whereas wild-type C. bescii lacks the ability to make ethanol, 70% of the fermentation products in the engineered strain were ethanol [12.8 mM ethanol directly from 2% (wt/vol) switchgrass, a real-world substrate] with decreased production of acetate by 38% compared with wild-type. Direct conversion of biomass to ethanol represents a new paradigm for consolidated bioprocessing, offering the potential for carbon neutral, cost-effective, sustainable fuel production.

  12. The desorption of Phosphorous (32 P) fixed on iron and aluminum oxy-hydroxide surfaces by the soil microbial biomass

    International Nuclear Information System (INIS)

    Araujo, Lilian Maria Cesar de.

    1995-02-01

    This work determines whether the soil microbial biomass, with an ample supply of available C, can utilize P adsorber in the surfaces of oxy-hydroxides of Fe or Al of soil-P deficient soils. To simulate the surfaces of the natural Fe and Al compounds, synthetic oxy-hydroxides of Fe and Al, impregnated in strips of filter paper, and containing P tagged with 32 P, were used. (author). 60 refs., 7 figs., 7 tabs

  13. Mapping and determinism of soil microbial community distribution across an agricultural landscape.

    Science.gov (United States)

    Constancias, Florentin; Terrat, Sébastien; Saby, Nicolas P A; Horrigue, Walid; Villerd, Jean; Guillemin, Jean-Philippe; Biju-Duval, Luc; Nowak, Virginie; Dequiedt, Samuel; Ranjard, Lionel; Chemidlin Prévost-Bouré, Nicolas

    2015-06-01

    Despite the relevance of landscape, regarding the spatial patterning of microbial communities and the relative influence of environmental parameters versus human activities, few investigations have been conducted at this scale. Here, we used a systematic grid to characterize the distribution of soil microbial communities at 278 sites across a monitored agricultural landscape of 13 km². Molecular microbial biomass was estimated by soil DNA recovery and bacterial diversity by 16S rRNA gene pyrosequencing. Geostatistics provided the first maps of microbial community at this scale and revealed a heterogeneous but spatially structured distribution of microbial biomass and diversity with patches of several hundreds of meters. Variance partitioning revealed that both microbial abundance and bacterial diversity distribution were highly dependent of soil properties and land use (total variance explained ranged between 55% and 78%). Microbial biomass and bacterial richness distributions were mainly explained by soil pH and texture whereas bacterial evenness distribution was mainly related to land management. Bacterial diversity (richness, evenness, and Shannon index) was positively influenced by cropping intensity and especially by soil tillage, resulting in spots of low microbial diversity in soils under forest management. Spatial descriptors also explained a small but significant portion of the microbial distribution suggesting that landscape configuration also shapes microbial biomass and bacterial diversity. © 2015 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  14. %22Trojan Horse%22 strategy for deconstruction of biomass for biofuels production.

    Energy Technology Data Exchange (ETDEWEB)

    Simmons, Blake Alexander; Sinclair, Michael B.; Yu, Eizadora; Timlin, Jerilyn Ann; Hadi, Masood Z.; Tran-Gyamfi, Mary

    2011-02-01

    Production of renewable biofuels to displace fossil fuels currently consumed in the transportation sector is a pressing multiagency national priority (DOE/USDA/EERE). Currently, nearly all fuel ethanol is produced from corn-derived starch. Dedicated 'energy crops' and agricultural waste are preferred long-term solutions for renewable, cheap, and globally available biofuels as they avoid some of the market pressures and secondary greenhouse gas emission challenges currently facing corn ethanol. These sources of lignocellulosic biomass are converted to fermentable sugars using a variety of chemical and thermochemical pretreatments, which disrupt cellulose and lignin cross-links, allowing exogenously added recombinant microbial enzymes to more efficiently hydrolyze the cellulose for 'deconstruction' into glucose. This process is plagued with inefficiencies, primarily due to the recalcitrance of cellulosic biomass, mass transfer issues during deconstruction, and low activity of recombinant deconstruction enzymes. Costs are also high due to the requirement for enzymes and reagents, and energy-intensive cumbersome pretreatment steps. One potential solution to these problems is found in synthetic biology-engineered plants that self-produce a suite of cellulase enzymes. Deconstruction can then be integrated into a one-step process, thereby increasing efficiency (cellulose-cellulase mass-transfer rates) and reducing costs. The unique aspects of our approach are the rationally engineered enzymes which become Trojan horses during pretreatment conditions. During this study we rationally engineered Cazy enzymes and then integrated them into plant cells by multiple transformation techniques. The regenerated plants were assayed for first expression of these messages and then for the resulting proteins. The plants were then subjected to consolidated bioprocessing and characterized in detail. Our results and possible implications of this work on developing

  15. Oxidoreductive Cellulose Depolymerization by the Enzymes Cellobiose Dehydrogenase and Glycoside Hydrolase 61▿†

    Science.gov (United States)

    Langston, James A.; Shaghasi, Tarana; Abbate, Eric; Xu, Feng; Vlasenko, Elena; Sweeney, Matt D.

    2011-01-01

    Several members of the glycoside hydrolase 61 (GH61) family of proteins have recently been shown to dramatically increase the breakdown of lignocellulosic biomass by microbial hydrolytic cellulases. However, purified GH61 proteins have neither demonstrable direct hydrolase activity on various polysaccharide or lignacious components of biomass nor an apparent hydrolase active site. Cellobiose dehydrogenase (CDH) is a secreted flavocytochrome produced by many cellulose-degrading fungi with no well-understood biological function. Here we demonstrate that the binary combination of Thermoascus aurantiacus GH61A (TaGH61A) and Humicola insolens CDH (HiCDH) cleaves cellulose into soluble, oxidized oligosaccharides. TaGH61A-HiCDH activity on cellulose is shown to be nonredundant with the activities of canonical endocellulase and exocellulase enzymes in microcrystalline cellulose cleavage, and while the combination of TaGH61A and HiCDH cleaves highly crystalline bacterial cellulose, it does not cleave soluble cellodextrins. GH61 and CDH proteins are coexpressed and secreted by the thermophilic ascomycete Thielavia terrestris in response to environmental cellulose, and the combined activities of T. terrestris GH61 and T. terrestris CDH are shown to synergize with T. terrestris cellulose hydrolases in the breakdown of cellulose. The action of GH61 and CDH on cellulose may constitute an important, but overlooked, biological oxidoreductive system that functions in microbial lignocellulose degradation and has applications in industrial biomass utilization. PMID:21821740

  16. A Review of the Role of Amphiphiles in Biomass to Ethanol Conversion

    Directory of Open Access Journals (Sweden)

    William Gibbons

    2013-04-01

    Full Text Available One of the concerns for economical production of ethanol from biomass is the large volume and high cost of the cellulolytic enzymes used to convert biomass into fermentable sugars. The presence of acetyl groups in hemicellulose and lignin in plant cell walls reduces accessibility of biomass to the enzymes and makes conversion a slow process. In addition to low enzyme accessibility, a rapid deactivation of cellulases during biomass hydrolysis can be another factor contributing to the low sugar recovery. As of now, the economical reduction in lignin content of the biomass is considered a bottleneck, and raises issues for several reasons. The presence of lignin in biomass reduces the swelling of cellulose fibrils and accessibility of enzyme to carbohydrate polymers. It also causes an irreversible adsorption of the cellulolytic enzymes that prevents effective enzyme activity and recycling. Amphiphiles, such as surfactants and proteins have been found to improve enzyme activity by several mechanisms of action that are not yet fully understood. Reduction in irreversible adsorption of enzyme to non-specific sites, reduction in viscosity of liquid and surface tension and consequently reduced contact of enzyme with air-liquid interface, and modifications in biomass chemical structure are some of the benefits derived from surface active molecules. Application of some of these amphiphiles could potentially reduce the capital and operating costs of bioethanol production by reducing fermentation time and the amount of enzyme used for saccharification of biomass. In this review article, the benefit of applying amphiphiles at various stages of ethanol production (i.e., pretreatment, hydrolysis and hydrolysis-fermentation is reviewed and the proposed mechanisms of actions are described.

  17. Mining anaerobic digester consortia metagenomes for secreted carbohydrate active enzymes

    DEFF Research Database (Denmark)

    Wilkens, Casper; Busk, Peter Kamp; Pilgaard, Bo

    thermophilic and mesophilic ADs a wide variety of carbohydrate active enzyme functions were discovered in the metagenomic sequencing of the microbial consortia. The most dominating type of glycoside hydrolases were β-glucosidases (up to 27%), α-amylases (up to 10%), α-glucosidases (up to 8%), α......, and food wastes (Alvarado et al., 2014). The processes and the roles of the microorganisms that are involved in biomass conversion and methane production in ADs are still not fully understood. We are investigating thermophilic and mesophilic ADs that use wastewater surplus sludge for methane production...... was done with the Peptide Pattern Recognition (PPR) program (Busk and Lange, 2013), which is a novel non-alignment based approach that can predict function of e.g. CAZymes. PPR identifies a set of short conserved sequences, which can be used as a finger print when mining genomes for novel enzymes. In both...

  18. Effects of Pulp and Na-Bentonite Amendments on the Mobility of Trace Elements, Soil Enzymes Activity and Microbial Parameters under Ex Situ Aided Phytostabilization

    Science.gov (United States)

    Wasilkowski, Daniel; Mrozik, Agnieszka

    2017-01-01

    The objective of this study was to explore the potential use of pulp (by-product) from coffee processing and Na-bentonite (commercial product) for minimizing the environmental risk of Zn, Pb and Cd in soil collected from a former mine and zinc-lead smelter. The effects of soil amendments on the physicochemical properties of soil, the structural and functional diversity of the soil microbiome as well as soil enzymes were investigated. Moreover, biomass of Festuca arundinacea Schreb. (cultivar Asterix) and the uptake of trace elements in plant tissues were studied. The outdoor pot set contained the following soils: control soil (initial), untreated soil (without additives) with grass cultivation and soils treated (with additives) with and without plant development. All of the selected parameters were measured at the beginning of the experiment (t0), after 2 months of chemical stabilization (t2) and at the end of the aided phytostabilization process (t14). The obtained results indicated that both amendments efficiently immobilized the bioavailable fractions of Zn (87–91%) and Cd (70–83%) at t14; however, they were characterized by a lower ability to bind Pb (33–50%). Pulp and Na-bentonite drastically increased the activity of dehydrogenase (70- and 12-fold, respectively) at t14, while the activities of urease, acid and alkaline phosphatases differed significantly depending on the type of material that was added into the soil. Generally, the activities of these enzymes increased; however, the increase was greater for pulp (3.5-6-fold) than for the Na-bentonite treatment (1.3–2.2-fold) as compared to the control. Soil additives significantly influenced the composition and dynamics of the soil microbial biomass over the experiment. At the end, the contribution of microbial groups could be ordered as follows: gram negative bacteria, fungi, gram positive bacteria, actinomycetes regardless of the type of soil enrichment. Conversely, the shift in the functional

  19. Effects of Pulp and Na-Bentonite Amendments on the Mobility of Trace Elements, Soil Enzymes Activity and Microbial Parameters under Ex Situ Aided Phytostabilization.

    Directory of Open Access Journals (Sweden)

    Daniel Wasilkowski

    Full Text Available The objective of this study was to explore the potential use of pulp (by-product from coffee processing and Na-bentonite (commercial product for minimizing the environmental risk of Zn, Pb and Cd in soil collected from a former mine and zinc-lead smelter. The effects of soil amendments on the physicochemical properties of soil, the structural and functional diversity of the soil microbiome as well as soil enzymes were investigated. Moreover, biomass of Festuca arundinacea Schreb. (cultivar Asterix and the uptake of trace elements in plant tissues were studied. The outdoor pot set contained the following soils: control soil (initial, untreated soil (without additives with grass cultivation and soils treated (with additives with and without plant development. All of the selected parameters were measured at the beginning of the experiment (t0, after 2 months of chemical stabilization (t2 and at the end of the aided phytostabilization process (t14. The obtained results indicated that both amendments efficiently immobilized the bioavailable fractions of Zn (87-91% and Cd (70-83% at t14; however, they were characterized by a lower ability to bind Pb (33-50%. Pulp and Na-bentonite drastically increased the activity of dehydrogenase (70- and 12-fold, respectively at t14, while the activities of urease, acid and alkaline phosphatases differed significantly depending on the type of material that was added into the soil. Generally, the activities of these enzymes increased; however, the increase was greater for pulp (3.5-6-fold than for the Na-bentonite treatment (1.3-2.2-fold as compared to the control. Soil additives significantly influenced the composition and dynamics of the soil microbial biomass over the experiment. At the end, the contribution of microbial groups could be ordered as follows: gram negative bacteria, fungi, gram positive bacteria, actinomycetes regardless of the type of soil enrichment. Conversely, the shift in the functional

  20. Co-solvent pretreatment reduces costly enzyme requirements for high sugar and ethanol yields from lignocellulosic biomass.

    Science.gov (United States)

    Nguyen, Thanh Yen; Cai, Charles M; Kumar, Rajeev; Wyman, Charles E

    2015-05-22

    We introduce a new pretreatment called co-solvent-enhanced lignocellulosic fractionation (CELF) to reduce enzyme costs dramatically for high sugar yields from hemicellulose and cellulose, which is essential for the low-cost conversion of biomass to fuels. CELF employs THF miscible with aqueous dilute acid to obtain up to 95 % theoretical yield of glucose, xylose, and arabinose from corn stover even if coupled with enzymatic hydrolysis at only 2 mgenzyme  gglucan (-1) . The unusually high saccharification with such low enzyme loadings can be attributed to a very high lignin removal, which is supported by compositional analysis, fractal kinetic modeling, and SEM imaging. Subsequently, nearly pure lignin product can be precipitated by the evaporation of volatile THF for recovery and recycling. Simultaneous saccharification and fermentation of CELF-pretreated solids with low enzyme loadings and Saccharomyces cerevisiae produced twice as much ethanol as that from dilute-acid-pretreated solids if both were optimized for corn stover. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Comparison of model microbial allocation parameters in soils of varying texture

    Science.gov (United States)

    Hagerty, S. B.; Slessarev, E.; Schimel, J.

    2017-12-01

    The soil microbial community decomposes the majority of carbon (C) inputs to the soil. However, not all of this C is respired—rather, a substantial portion of the carbon processed by microbes may remain stored in the soil. The balance between C storage and respiration is controlled by microbial turnover rates and C allocation strategies. These microbial community properties may depend on soil texture, which has the potential to influence both the nature and the fate of microbial necromass and extracellular products. To evaluate the role of texture on microbial turnover and C allocation, we sampled four soils from the University of California's Hastings Reserve that varied in texture (one silt loam, two sandy loam, and on clay soil), but support similar grassland plant communities. We added 14C- glucose to the soil and measured the concentration of the label in the carbon dioxide (CO2), microbial biomass, and extractable C pools over 7 weeks. The labeled biomass turned over the slowest in the clay soil; the concentration of labeled biomass was more than 1.5 times the concentration of the other soils after 8 weeks. The clay soil also had the lowest mineralization rate of the label, and mineralization slowed after two weeks. In contrast, in the sandier soils mineralization rates were higher and did not plateau until 5 weeks into the incubation period. We fit the 14C data to a microbial allocation model and estimated microbial parameters; assimilation efficiency, exudation, and biomass specific respiration and turnover for each soil. We compare these parameters across the soil texture gradient to assess the extent to which models may need to account for variability in microbial C allocation across soils of different texture. Our results suggest that microbial C turns over more slowly in high-clay soils than in sandy soils, and that C lost from microbial biomass is retained at higher rates in high-clay soils. Accounting for these differences in microbial allocation

  2. BioDry: An Inexpensive, Low-Power Method to Preserve Aquatic Microbial Biomass at Room Temperature.

    Science.gov (United States)

    Tuorto, Steven J; Brown, Chris M; Bidle, Kay D; McGuinness, Lora R; Kerkhof, Lee J

    2015-01-01

    This report describes BioDry (patent pending), a method for reliably preserving the biomolecules associated with aquatic microbial biomass samples, without the need of hazardous materials (e.g. liquid nitrogen, preservatives, etc.), freezing, or bulky storage/sampling equipment. Gel electrophoresis analysis of nucleic acid extracts from samples treated in the lab with the BioDry method indicated that molecular integrity was protected in samples stored at room temperature for up to 30 days. Analysis of 16S/18S rRNA genes for presence/absence and relative abundance of microorganisms using both 454-pyrosequencing and TRFLP profiling revealed statistically indistinguishable communities from control samples that were frozen in liquid nitrogen immediately after collection. Seawater and river water biomass samples collected with a portable BioDry "field unit", constructed from off-the-shelf materials and a battery-operated pumping system, also displayed high levels of community rRNA preservation, despite a slight decrease in nucleic acid recovery over the course of storage for 30 days. Functional mRNA and protein pools from the field samples were also effectively conserved with BioDry, as assessed by respective RT-PCR amplification and western blot of ribulose-1-5-bisphosphate carboxylase/oxygenase. Collectively, these results demonstrate that BioDry can adequately preserve a suite of biomolecules from aquatic biomass at ambient temperatures for up to a month, giving it great potential for high resolution sampling in remote locations or on autonomous platforms where space and power are limited.

  3. BioDry: An Inexpensive, Low-Power Method to Preserve Aquatic Microbial Biomass at Room Temperature.

    Directory of Open Access Journals (Sweden)

    Steven J Tuorto

    Full Text Available This report describes BioDry (patent pending, a method for reliably preserving the biomolecules associated with aquatic microbial biomass samples, without the need of hazardous materials (e.g. liquid nitrogen, preservatives, etc., freezing, or bulky storage/sampling equipment. Gel electrophoresis analysis of nucleic acid extracts from samples treated in the lab with the BioDry method indicated that molecular integrity was protected in samples stored at room temperature for up to 30 days. Analysis of 16S/18S rRNA genes for presence/absence and relative abundance of microorganisms using both 454-pyrosequencing and TRFLP profiling revealed statistically indistinguishable communities from control samples that were frozen in liquid nitrogen immediately after collection. Seawater and river water biomass samples collected with a portable BioDry "field unit", constructed from off-the-shelf materials and a battery-operated pumping system, also displayed high levels of community rRNA preservation, despite a slight decrease in nucleic acid recovery over the course of storage for 30 days. Functional mRNA and protein pools from the field samples were also effectively conserved with BioDry, as assessed by respective RT-PCR amplification and western blot of ribulose-1-5-bisphosphate carboxylase/oxygenase. Collectively, these results demonstrate that BioDry can adequately preserve a suite of biomolecules from aquatic biomass at ambient temperatures for up to a month, giving it great potential for high resolution sampling in remote locations or on autonomous platforms where space and power are limited.

  4. Detection of enzyme activity in decontaminated spices of industrial use

    International Nuclear Information System (INIS)

    Müller, R.; Theobald, R.

    1995-01-01

    A range of decontaminated spices of industrial use have been examinated for their enzymes (catalase, peroxidase, amylase, lipase activity). The genuine enzymes remain fully active in irradiated spices, whereas the microbial load is clearly reduced. In contrast steam treated spices no longer demonstrate enzyme activities. Steam treatment offers e.g. black pepper without lipase activity, which can no longer cause fat deterioration. Low microbial load in combination with clearly detectable enzyme activity in spices is an indication for irradiation, whereas, reduced microbial contamination combined with enzyme inactivation indicate steam treatment of raw material [de

  5. Transporter engineering in biomass utilization by yeast.

    Science.gov (United States)

    Hara, Kiyotaka Y; Kobayashi, Jyumpei; Yamada, Ryosuke; Sasaki, Daisuke; Kuriya, Yuki; Hirono-Hara, Yoko; Ishii, Jun; Araki, Michihiro; Kondo, Akihiko

    2017-11-01

    Biomass resources are attractive carbon sources for bioproduction because of their sustainability. Many studies have been performed using biomass resources to produce sugars as carbon sources for cell factories. Expression of biomass hydrolyzing enzymes in cell factories is an important approach for constructing biomass-utilizing bioprocesses because external addition of these enzymes is expensive. In particular, yeasts have been extensively engineered to be cell factories that directly utilize biomass because of their manageable responses to many genetic engineering tools, such as gene expression, deletion and editing. Biomass utilizing bioprocesses have also been developed using these genetic engineering tools to construct metabolic pathways. However, sugar input and product output from these cells are critical factors for improving bioproduction along with biomass utilization and metabolic pathways. Transporters are key components for efficient input and output activities. In this review, we focus on transporter engineering in yeast to enhance bioproduction from biomass resources. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  6. Analysis of soil microbial community structure and enzyme activities associated with negative effects of pseudostellaria heterophylla consecutive monoculture on yield

    International Nuclear Information System (INIS)

    Lin, S.; Lin, W.X.

    2015-01-01

    Pseudostellaria heterophylla is an important medicinal plant in China. However, cultivation of P. heterophylla using consecutive monoculture results in significant reductions in yield and quality. In this study, terminal-restriction fragment length polymorphism (T-RFLP) analysis and measurement of soil enzyme activities were used to investigate the regulation of soil micro-ecology to identify ways to overcome the negative effects of P. heterophylla consecutive monoculture. T-RFLP analysis showed that rice/P. heterophylla (RP) and bean/P. heterophylla (BP) crop rotation systems increased the number and diversity of microbial groups in P. heterophylla rhizosphere soil. In particular, the RP and BP crop rotations increased the number and abundance of beneficial bacterial species compared with two-year consecutive monoculture of P. heterophylla. The presence of these beneficial bacteria was positively correlated with soil enzyme activities which increased in rhizosphere soils of the RP and BP crop rotation systems. The results indicated that crop rotation systems could increase activities of key soil enzymes and beneficial microbial groups and improve soil health. This study could provide a theoretical basis to resolve the problems associated with P. heterophylla consecutive monoculture. (author)

  7. Microbial-processing of fruit and vegetable wastes for production of vital enzymes and organic acids: Biotechnology and scopes.

    Science.gov (United States)

    Panda, Sandeep K; Mishra, Swati S; Kayitesi, Eugenie; Ray, Ramesh C

    2016-04-01

    Wastes generated from fruits and vegetables are organic in nature and contribute a major share in soil and water pollution. Also, green house gas emission caused by fruit and vegetable wastes (FVWs) is a matter of serious environmental concern. This review addresses the developments over the last one decade on microbial processing technologies for production of enzymes and organic acids from FVWs. The advances in genetic engineering for improvement of microbial strains in order to enhance the production of the value added bio-products as well as the concept of zero-waste economy have been briefly discussed. Copyright © 2015 Elsevier Inc. All rights reserved.

  8. Enzyme Characterization of Cellulase and Hemicellulases Component Enzymes and Saccharification of Ionic Liquid Pretreated Lignocellulosic Biomass

    Science.gov (United States)

    Lignocellulosic biomass is comprised of cellulose and hemicellulose, sources of polysaccharides, and lignin, a macromolecule with extensive aromaticity. Terrestrial biomass can provide a renewable carbon based feedstock for fuel and chemical production. However, recalcitrance of biomass to deconstru...

  9. Fiscal 2000 project of inviting proposals for international joint research - invitation for international proposal (Power generation No.18). Achievement report on development of device for monitoring biomass decomposing enzyme gene expression; 2000 nendo kokusai kyodo kenkyu teian kobo jigyo - kokusai teian kobo (hatsuden No.18). Biomass bunkai koso idenshi hatsugen monitoring device no kaihatsu seika hokokusho

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2001-03-01

    A fuel production system is under development to supply fuel for power generation, in which biomass is efficiently fermented. In this connection, based on biomass decomposing enzyme related gene information provided by Japan and the U.S., a DNA (deoxyribonucleic acid) chip is built, wherein biomass decomposing enzymes, control factors, and enzyme secretion enhancing genes are densely deposited. Efforts are made, using the chip for the monitoring of the enzyme production process during the culture of mold, to develop technologies for collecting information simultaneously and comprehensively concerning the expression of great numbers of biomass decomposing enzyme genes and concerning other genes that control the production or enhance the secretion of enzymes. In concrete terms, approximately 2000 types of genes are selected out of the Aspergillus EST database held by the Japanese and U.S. members, and cDNA from the original clones is amplified and purified for each DNA of the 2000 genes. The purified DNA is spotted and fixed on the glass plate for a DNA chip and applied to the mRNA pool whose gene expression profile is known, and the detection conditions are studied for the DNA chip and specifications are optimized. (NEDO)

  10. Soil Microbial Community Structure Evolution along Halophyte Succession in Bohai Bay Wetland

    Directory of Open Access Journals (Sweden)

    Mingyang Cong

    2014-01-01

    Full Text Available It is urgent to recover Bohai Bay costal wetland ecosystem because of covering a large area of severe saline-alkali soil. To explore the relationship between halophyte herbaceous succession and microbial community structure, we chose four local communities which played an important role in improving soil microenvironment. We performed phospholipid fatty acid analysis, measured soil parameters, and evaluated shifts of microbial community structure. Results showed that microbial community structure changed significantly along succession and bacteria community was dominant. Total phospholipid fatty acid content increased in different successional stages but decreased with depth, with similar variations in bacterial and fungal biomass. Soil organic carbon and especially total nitrogen were positively correlated with microbial biomass. Colonization of pioneering salt-tolerant plants Suaeda glauca in saline-alkali bare land changed total soil microorganism content and composition. These results showed that belowground processes were strongly related with aboveground halophyte succession. Fungal/bacterial ratio, Gram-negative/Gram-positive bacteria ratio, total microbial biomass, and fungi and bacteria content could indicate the degree of succession stages in Bohai Bay wetland ecosystem. And also these findings demonstrated that microbial community biomass and composition evolved along with vegetation succession environmental variables.

  11. Treatment of biomass to obtain fermentable sugars

    Science.gov (United States)

    Dunson, Jr., James B.; Tucker, Melvin [Lakewood, CO; Elander, Richard [Evergreen, CO; Hennessey, Susan M [Avondale, PA

    2011-04-26

    Biomass is pretreated using a low concentration of aqueous ammonia at high biomass concentration. Pretreated biomass is further hydrolyzed with a saccharification enzyme consortium. Fermentable sugars released by saccharification may be utilized for the production of target chemicals by fermentation.

  12. [Soil soluble organic matter, microbial biomass, and enzyme activities in forest plantations in degraded red soil region of Jiangxi Province, China].

    Science.gov (United States)

    Jiang, Yu-mei; Chen, Cheng-long; Xu, Zhi-hong; Liu, Yuan-qiu; Ouyang, Jing; Wang, Fang

    2010-09-01

    Taking the adjacent 18-year-old pure Pinus massoniana pure forest (I), P. massoniana, Liquidamber fomosana, and Schima superba mixed forest (II), S. superba pure forest (III), L. fomosana (IV) pure forest, and natural restoration fallow land (CK) in Taihe County of Jiangxi Province as test sites, a comparative study was made on their soil soluble organic carbon (SOC) and nitrogen (SON), soil microbial biomass C (MBC) and N (MBN), and soil urease and asparaginase activities. In 0-10 cm soil layer, the pool sizes of SOC, SON, MBC, and MBN at test sites ranged in 354-1007 mg x kg(-1), 24-73 mg x kg(-1), 203-488 mg x kg(-1), and 24-65 mg x kg(-1), and the soil urease and asparaginase activities were 95-133 mg x kg(-1) x d(-1) and 58-113 mg x kg(-1) x d(-1), respectively. There were significant differences in the pool sizes of SOC, SON, MBC, and MBN and the asparaginase activity among the test sites, but no significant difference was observed in the urease activity. The pool sizes of SOC and SON were in the order of IV > CK > III > I > II, those of MBC and MBN were in the order of CK > IV > III > I > II, and asparaginase activity followed the order of IV > CK > III > II > I. With the increase of soil depth, the pool sizes of SOC, SON, MBC, and MBN and the activities of soil asparaginase and urease decreased. In 0-20 cm soil layer, the SOC, SON, MBC, MBN, total C, and total N were highly correlated with each other, soil asparaginase activity was highly correlated with SOC, SON, TSN, total C, total N, MBC, and MBN, and soil urease activity was highly correlated with SON, TSN, total C, MBC and MBN.

  13. Response of the soil microbial community and soil nutrient bioavailability to biomass harvesting and reserve tree retention in northern Minnesota aspen-dominated forests

    Science.gov (United States)

    Tera E. Lewandowski; Jodi A. Forrester; David J. Mladenoff; Anthony W. D' Amato; Brian J. Palik

    2016-01-01

    Intensive forest biomass harvesting, or the removal of harvesting slash (woody debris from tree branches and tops) for use as biofuel, has the potential to negatively affect the soil microbial community (SMC) due to loss of carbon and nutrient inputs from the slash, alteration of the soil microclimate, and increased nutrient leaching. These effects could result in...

  14. Increased production of biomass-degrading enzymes by double deletion of creA and creB genes involved in carbon catabolite repression in Aspergillus oryzae.

    Science.gov (United States)

    Ichinose, Sakurako; Tanaka, Mizuki; Shintani, Takahiro; Gomi, Katsuya

    2018-02-01

    In a previous study, we reported that a double gene deletion mutant for CreA and CreB, which constitute the regulatory machinery involved in carbon catabolite repression, exhibited improved production of α-amylase compared with the wild-type strain and single creA or creB deletion mutants in Aspergillus oryzae. Because A. oryzae can also produce biomass-degrading enzymes, such as xylolytic and cellulolytic enzymes, we examined the production levels of those enzymes in deletion mutants in this study. Xylanase and β-glucosidase activities in the wild-type were hardly detected in submerged culture containing xylose as the carbon source, whereas those enzyme activities were significantly increased in the single creA deletion (ΔcreA) and double creA and creB deletion (ΔcreAΔcreB) mutants. In particular, the ΔcreAΔcreB mutant exhibited >100-fold higher xylanase and β-glucosidase activities than the wild-type. Moreover, in solid-state culture, the β-glucosidase activity of the double deletion mutant was >7-fold higher than in the wild-type. These results suggested that deletion of both creA and creB genes could also efficiently improve the production levels of biomass-degrading enzymes in A. oryzae. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  15. Linking genes to microbial growth kinetics: an integrated biochemical systems engineering approach.

    Science.gov (United States)

    Koutinas, Michalis; Kiparissides, Alexandros; Silva-Rocha, Rafael; Lam, Ming-Chi; Martins Dos Santos, Vitor A P; de Lorenzo, Victor; Pistikopoulos, Efstratios N; Mantalaris, Athanasios

    2011-07-01

    The majority of models describing the kinetic properties of a microorganism for a given substrate are unstructured and empirical. They are formulated in this manner so that the complex mechanism of cell growth is simplified. Herein, a novel approach for modelling microbial growth kinetics is proposed, linking biomass growth and substrate consumption rates to the gene regulatory programmes that control these processes. A dynamic model of the TOL (pWW0) plasmid of Pseudomonas putida mt-2 has been developed, describing the molecular interactions that lead to the transcription of the upper and meta operons, known to produce the enzymes for the oxidative catabolism of m-xylene. The genetic circuit model was combined with a growth kinetic model decoupling biomass growth and substrate consumption rates, which are expressed as independent functions of the rate-limiting enzymes produced by the operons. Estimation of model parameters and validation of the model's predictive capability were successfully performed in batch cultures of mt-2 fed with different concentrations of m-xylene, as confirmed by relative mRNA concentration measurements of the promoters encoded in TOL. The growth formation and substrate utilisation patterns could not be accurately described by traditional Monod-type models for a wide range of conditions, demonstrating the critical importance of gene regulation for the development of advanced models closely predicting complex bioprocesses. In contrast, the proposed strategy, which utilises quantitative information pertaining to upstream molecular events that control the production of rate-limiting enzymes, predicts the catabolism of a substrate and biomass formation and could be of central importance for the design of optimal bioprocesses. Copyright © 2011 Elsevier Inc. All rights reserved.

  16. [Effects of controlled release blend bulk urea on soil nitrogen and soil enzyme activity in wheat and rice fields].

    Science.gov (United States)

    Zhang, Jing Sheng; Wang, Chang Quan; Li, Bing; Liang, Jing Yue; He, Jie; Xiang, Hao; Yin, Bin; Luo, Jing

    2017-06-18

    A field experiment was conducted to investigate the effect of controlled-release fertilizer (CRF) combined with urea (UR) on the soil fertility and environment in wheat-rice rotation system. Changes in four forms of nitrogen (total nitrogen, ammonium nitrogen, nitrate nitrogen, and microbial biomass nitrogen) and in activities of three soil enzymes participating in nitrogen transformation (urease, protease, and nitrate reductase) were measured in seven fertilization treatments (no fertilization, routine fertilization, 10%CRF+90%UR, 20%CRF+80%UR, 40%CRF+60%UR, 80%CRF+20%UR, and 100%CRF). The results showed that soil total nitrogen was stable in the whole growth period of wheat and rice. There was no significant difference among the treatments of over 20% CRF in soil total nitrogen content of wheat and rice. The soil inorganic nitrogen content was increased dramatically in treatments of 40% or above CRF during the mid-late growing stages of wheat and rice. With the advance of the growth period, conventional fertilization significantly decreased soil microbial biomass nitrogen, but the treatments of 40% and above CRF increased the soil microbial biomass nitrogen significantly. The soil enzyme activities were increased with over 40% of CRF in the mid-late growing stage of wheat and rice. By increasing the CRF ratio, the soil protease activity and nitrate reductase activity were improved gradually, and peaked in 100% CRF. The treatments of above 20% CRF could decrease the urease activity in tillering stage of rice and delay the peak of ammonium nitrogen, which would benefit nitrogen loss reduction. The treatments of 40% and above CRF were beneficial to improving soil nitrogen supply and enhancing soil urease and protease activities, which could promote the effectiveness of nitrogen during the later growth stages of wheat and rice. The 100% CRF treatment improved the nitrate reductase activity significantly during the later stage of wheat and rice. Compared with the

  17. Production of xylitol from biomass using an inhibitor-tolerant fungal strain

    Science.gov (United States)

    Inhibitory compounds arising from physical–chemical pretreatment of biomass feedstock can interfere with fermentation of biomass sugars to product. A fungus, Coniochaeta ligniaria NRRL30616 improves fermentability of biomass sugars by metabolizing a variety of microbial inhibitors including furan al...

  18. Experimental methods for laboratory-scale ensilage of lignocellulosic biomass

    International Nuclear Information System (INIS)

    Tanjore, Deepti; Richard, Tom L.; Marshall, Megan N.

    2012-01-01

    Anaerobic fermentation is a potential storage method for lignocellulosic biomass in biofuel production processes. Since biomass is seasonally harvested, stocks are often dried or frozen at laboratory scale prior to fermentation experiments. Such treatments prior to fermentation studies cause irreversible changes in the plant cells, influencing the initial state of biomass and thereby the progression of the fermentation processes itself. This study investigated the effects of drying, refrigeration, and freezing relative to freshly harvested corn stover in lab-scale ensilage studies. Particle sizes, as well as post-ensilage drying temperatures for compositional analysis, were tested to identify the appropriate sample processing methods. After 21 days of ensilage the lowest pH value (3.73 ± 0.03), lowest dry matter loss (4.28 ± 0.26 g. 100 g-1DM), and highest water soluble carbohydrate (WSC) concentrations (7.73 ± 0.26 g. 100 g-1DM) were observed in control biomass (stover ensiled within 12 h of harvest without any treatments). WSC concentration was significantly reduced in samples refrigerated for 7 days prior to ensilage (3.86 ± 0.49 g. 100 g −1 DM). However, biomass frozen prior to ensilage produced statistically similar results to the fresh biomass control, especially in treatments with cell wall degrading enzymes. Grinding to decrease particle size reduced the variance amongst replicates for pH values of individual reactors to a minor extent. Drying biomass prior to extraction of WSCs resulted in degradation of the carbohydrates and a reduced estimate of their concentrations. The methods developed in this study can be used to improve ensilage experiments and thereby help in developing ensilage as a storage method for biofuel production. -- Highlights: ► Laboratory-scale methods to assess the influence of ensilage biofuel production. ► Drying, freezing, and refrigeration of biomass influenced microbial fermentation. ► Freshly ensiled stover exhibited

  19. Medium activity long-lived nuclear waste; microbial paradise or hadean environment - Evaluation of biomass and impact on redox conditions

    International Nuclear Information System (INIS)

    Albrecht, A.; Libert, M.

    2010-01-01

    near-field biogeochemistry without detailed knowledge of kinetic parameters: The first approach is a mass balance which considers the masses of both the oxidizing and the reducing agents in a single waste cell. In the simplest case only nitrates, sulphates and Fe(0) are considered; nitrates are reduced to N 2 , sulphates to S 2- , and all Fe is oxidised to Fe 3 O 4 , the latter generating hydrogen gas. Of the produced H 2 , 25% serves to reduce nitrates and 55% to reduce sulphates. Further mass balance calculations are underway that consider the direct reaction of nitrates with Fe(0), a reaction for which very little relevant information is available. It is know that nitrate solutions may inhibit corrosion, but they are also known to react with iron. Furthermore, Fe(III), initially present in the engineered barrier or in the host rock and produced as a consequence of Fe(II) oxidation by O 2 during waste cell opening, will also be considered. The second approach combines the previous one with a quantification of free energies, which in turn can be used to form microbial biomass according to the simple relationship 1 g per 64 kJ. With balanced stoichiometric reactions and thermodynamic data at pH 12 first estimates indicate a production of 0.7 kg of biomass per waste container. This is significantly higher than what has been estimated for high-level waste cells, where bacterial activity was modelled to be essentially limited to the first 100 years, when the system is partially characterised by the presence of O 2 . However, these preliminary calculations assume that all nutrients are simultaneously bioavailable and that the energy requirement for microbial biomass production at low pH can be used as a first estimate for higher pH setting. Ongoing work will be presented that considers parameter uncertainties using a Monte Carlo approach, results which allow assessing the upper and lower limits of bacterial production. These values can then be used to quantify the

  20. Bioprospecting metagenomics of decaying wood: mining for new glycoside hydrolases

    Directory of Open Access Journals (Sweden)

    Li Luen-Luen

    2011-08-01

    Full Text Available Abstract Background To efficiently deconstruct recalcitrant plant biomass to fermentable sugars in industrial processes, biocatalysts of higher performance and lower cost are required. The genetic diversity found in the metagenomes of natural microbial biomass decay communities may harbor such enzymes. Our goal was to discover and characterize new glycoside hydrolases (GHases from microbial biomass decay communities, especially those from unknown or never previously cultivated microorganisms. Results From the metagenome sequences of an anaerobic microbial community actively decaying poplar biomass, we identified approximately 4,000 GHase homologs. Based on homology to GHase families/activities of interest and the quality of the sequences, candidates were selected for full-length cloning and subsequent expression. As an alternative strategy, a metagenome expression library was constructed and screened for GHase activities. These combined efforts resulted in the cloning of four novel GHases that could be successfully expressed in Escherichia coli. Further characterization showed that two enzymes showed significant activity on p-nitrophenyl-α-L-arabinofuranoside, one enzyme had significant activity against p-nitrophenyl-β-D-glucopyranoside, and one enzyme showed significant activity against p-nitrophenyl-β-D-xylopyranoside. Enzymes were also tested in the presence of ionic liquids. Conclusions Metagenomics provides a good resource for mining novel biomass degrading enzymes and for screening of cellulolytic enzyme activities. The four GHases that were cloned may have potential application for deconstruction of biomass pretreated with ionic liquids, as they remain active in the presence of up to 20% ionic liquid (except for 1-ethyl-3-methylimidazolium diethyl phosphate. Alternatively, ionic liquids might be used to immobilize or stabilize these enzymes for minimal solvent processing of biomass.

  1. High activity and low temperature optima of extracellular enzymes in Arctic sediments: implications for carbon cycling by heterotrophic microbial communities

    DEFF Research Database (Denmark)

    Arnosti, C.; Jørgensen, BB

    2003-01-01

    The rate of the initial step in microbial remineralization of organic carbon, extracellular enzymatic hydrolysis, was investigated as a function of temperature in permanently cold sediments from 2 fjords on the west coast of Svalbard (Arctic Ocean). We used 4 structurally distinct polysaccharides...... hydrolysis in order to determine the relative temperature responses of the initial and terminal steps in microbial remineralization of carbon. The temperature optimum of sulfate reduction, 21degreesC, was considerably lower than previous reports of sulfate reduction in marine sediments, but is consistent...... with recent studies of psychrophilic sulfate reducers isolated from Svalbard sediments. A calculation of potential carbon flow into the microbial food chain demonstrated that the activity of just one type of polysaccharide-hydrolyzing enzyme could in theory supply 21 to 100% of the carbon consumed via sulfate...

  2. Sedimentary organic biomarkers suggest detrimental effects of PAHs on estuarine microbial biomass during the 20th century in San Francisco Bay, CA, USA

    Science.gov (United States)

    Nilsen, Elena B.; Rosenbauer, Robert J.; Fuller, Christopher C.; Jaffe, Bruce E.

    2014-01-01

    Hydrocarbon contaminants are ubiquitous in urban aquatic ecosystems, and the ability of some microbial strains to degrade certain polycyclic aromatic hydrocarbons (PAHs) is well established. However, detrimental effects of petroleum hydrocarbon contamination on nondegrader microbial populations and photosynthetic organisms have not often been considered. In the current study, fatty acid methyl ester (FAME) biomarkers in the sediment record were used to assess historical impacts of petroleum contamination on microbial and/or algal biomass in South San Francisco Bay, CA, USA. Profiles of saturated, branched, and monounsaturated fatty acids had similar concentrations and patterns downcore. Total PAHs in a sediment core were on average greater than 20× higher above ∼200 cm than below, which corresponds roughly to the year 1900. Isomer ratios were consistent with a predominant petroleum combustion source for PAHs. Several individual PAHs exceeded sediment quality screening values. Negative correlations between petroleum contaminants and microbial and algal biomarkers – along with high trans/cis ratios of unsaturated FA, and principle component analysis of the PAH and fatty acid records – suggest a negative impacts of petroleum contamination, appearing early in the 20th century, on microbial and/or algal ecology at the site.

  3. Microbial response of an acid forest soil to experimental soil warming

    Science.gov (United States)

    S.S. Arnold; I.J. Fernandez; L.E. Rustad; L.M. Zibilske

    1999-01-01

    Effects of increased soil temperature on soil microbial biomass and dehydrogenase activity were examined on organic (O) horizon material in a low-elevation spruce-fir ecosystem. Soil temperature was maintained at 5 °C above ambient during the growing season in the experimental plots, and soil temperature, moisture, microbial biomass, and dehydrogenase activity were...

  4. Dynamics of soil organic carbon and microbial biomass carbon in relation to water erosion and tillage erosion.

    Science.gov (United States)

    Xiaojun, Nie; Jianhui, Zhang; Zhengan, Su

    2013-01-01

    Dynamics of soil organic carbon (SOC) are associated with soil erosion, yet there is a shortage of research concerning the relationship between soil erosion, SOC, and especially microbial biomass carbon (MBC). In this paper, we selected two typical slope landscapes including gentle and steep slopes from the Sichuan Basin, China, and used the (137)Cs technique to determine the effects of water erosion and tillage erosion on the dynamics of SOC and MBC. Soil samples for the determination of (137)Cs, SOC, MBC and soil particle-size fractions were collected on two types of contrasting hillslopes. (137)Cs data revealed that soil loss occurred at upper slope positions of the two landscapes and soil accumulation at the lower slope positions. Soil erosion rates as well as distribution patterns of the erosion is the major process of soil redistribution in the gentle slope landscape, while tillage erosion acts as the dominant process of soil redistribution in the steep slope landscape. In gentle slope landscapes, both SOC and MBC contents increased downslope and these distribution patterns were closely linked to soil redistribution rates. In steep slope landscapes, only SOC contents increased downslope, dependent on soil redistribution. It is noticeable that MBC/SOC ratios were significantly lower in gentle slope landscapes than in steep slope landscapes, implying that water erosion has a negative effect on the microbial biomass compared with tillage erosion. It is suggested that MBC dynamics are closely associated with soil redistribution by water erosion but independent of that by tillage erosion, while SOC dynamics are influenced by soil redistribution by both water erosion and tillage erosion.

  5. Metagenomic analysis of a tropical composting operation at the são paulo zoo park reveals diversity of biomass degradation functions and organisms.

    Directory of Open Access Journals (Sweden)

    Layla Farage Martins

    Full Text Available Composting operations are a rich source for prospection of biomass degradation enzymes. We have analyzed the microbiomes of two composting samples collected in a facility inside the São Paulo Zoo Park, in Brazil. All organic waste produced in the park is processed in this facility, at a rate of four tons/day. Total DNA was extracted and sequenced with Roche/454 technology, generating about 3 million reads per sample. To our knowledge this work is the first report of a composting whole-microbial community using high-throughput sequencing and analysis. The phylogenetic profiles of the two microbiomes analyzed are quite different, with a clear dominance of members of the Lactobacillus genus in one of them. We found a general agreement of the distribution of functional categories in the Zoo compost metagenomes compared with seven selected public metagenomes of biomass deconstruction environments, indicating the potential for different bacterial communities to provide alternative mechanisms for the same functional purposes. Our results indicate that biomass degradation in this composting process, including deconstruction of recalcitrant lignocellulose, is fully performed by bacterial enzymes, most likely by members of the Clostridiales and Actinomycetales orders.

  6. Soil Microbial Activity in Conventional and Organic Agricultural Systems

    Directory of Open Access Journals (Sweden)

    Romero F.V. Carneiro

    2009-06-01

    Full Text Available The aim of this study was to evaluate microbial activity in soils under conventional and organic agricultural system management regimes. Soil samples were collected from plots under conventional management (CNV, organic management (ORG and native vegetation (AVN. Soil microbial activity and biomass was significantly greater in ORG compared with CNV. Soil bulk density decreased three years after adoption of organic system. Soil organic carbon (SOC was higher in the ORG than in the CNV. The soil under organic agricultural system presents higher microbial activity and biomass and lower bulk density than the conventional agricultural system.

  7. Switchgrass ecotypes alter microbial contribution to deep-soil C

    Science.gov (United States)

    Roosendaal, Damaris; Stewart, Catherine E.; Denef, Karolien; Follett, Ronald F.; Pruessner, Elizabeth; Comas, Louise H.; Varvel, Gary E.; Saathoff, Aaron; Palmer, Nathan; Sarath, Gautam; Jin, Virginia L.; Schmer, Marty; Soundararajan, Madhavan

    2016-05-01

    Switchgrass (Panicum virgatum L.) is a C4, perennial grass that is being developed as a bioenergy crop for the United States. While aboveground biomass production is well documented for switchgrass ecotypes (lowland, upland), little is known about the impact of plant belowground productivity on microbial communities down deep in the soil profiles. Microbial dynamics in deeper soils are likely to exert considerable control on ecosystem services, including C and nutrient cycles, due to their involvement in such processes as soil formation and ecosystem biogeochemistry. Differences in root biomass and rooting characteristics of switchgrass ecotypes could lead to distinct differences in belowground microbial biomass and microbial community composition. We quantified root abundance and root architecture and the associated microbial abundance, composition, and rhizodeposit C uptake for two switchgrass ecotypes using stable-isotope probing of microbial phospholipid fatty acids (PLFAs) after 13CO2 pulse-chase labeling. Kanlow, a lowland ecotype with thicker roots, had greater plant biomass above- and belowground (g m-2), greater root mass density (mg cm-3), and lower specific root length (m g-1) compared to Summer, an upland ecotype with finer root architecture. The relative abundance of bacterial biomarkers dominated microbial PLFA profiles for soils under both Kanlow and Summer (55.4 and 53.5 %, respectively; P = 0.0367), with differences attributable to a greater relative abundance of Gram-negative bacteria in soils under Kanlow (18.1 %) compared to soils under Summer (16.3 %; P = 0.0455). The two ecotypes also had distinctly different microbial communities process rhizodeposit C: greater relative atom % 13C excess in Gram-negative bacteria (44.1 ± 2.3 %) under the thicker roots of Kanlow and greater relative atom % 13C excess in saprotrophic fungi under the thinner roots of Summer (48.5 ± 2.2 %). For bioenergy production systems, variation between switchgrass

  8. Microbial biomass and carbon mineralization in agricultural soils as affected by pesticide addition.

    Science.gov (United States)

    Kumar, Anjani; Nayak, A K; Shukla, Arvind K; Panda, B B; Raja, R; Shahid, Mohammad; Tripathi, Rahul; Mohanty, Sangita; Rath, P C

    2012-04-01

    A laboratory study was conducted with four pesticides, viz. a fungicide (carbendazim), two insecticides (chlorpyrifos and cartap hydrochloride) and an herbicide (pretilachlor) applied to a sandy clay loam soil at a field rate to determine their effect on microbial biomass carbon (MBC) and carbon mineralization (C(min)). The MBC content of soil increased with time up to 30 days in cartap hydrochloride as well as chlorpyrifos treated soil. Thereafter, it decreased and reached close to the initial level by 90th day. However, in carbendazim treated soil, the MBC showed a decreasing trend up to 45 days and subsequently increased up to 90 days. In pretilachlor treated soil, MBC increased through the first 15 days, and thereafter decreased to the initial level. Application of carbendazim, chlorpyrifos and cartap hydrochloride decreased C(min) for the first 30 days and then increased afterwards, while pretilachlor treated soil showed an increasing trend.

  9. Characterization and mode of action of enzymes degrading galactan structures of arabinogalactans

    NARCIS (Netherlands)

    Vis, van de J.W.

    1994-01-01

    Agricultural biomass consisting mainly of cellulose, hemicellulose and lignin, is a renewable source of fuels and chemicals. An interesting option is enzymic conversion of biomass to readily usable material. To improve the overall economics of enzymic conversion of biomass not only

  10. A multi-substrate approach for functional metagenomics-based screening for (hemi)cellulases in two wheat straw-degrading microbial consortia unveils novel thermoalkaliphilic enzymes.

    Science.gov (United States)

    Maruthamuthu, Mukil; Jiménez, Diego Javier; Stevens, Patricia; van Elsas, Jan Dirk

    2016-01-28

    Functional metagenomics is a promising strategy for the exploration of the biocatalytic potential of microbiomes in order to uncover novel enzymes for industrial processes (e.g. biorefining or bleaching pulp). Most current methodologies used to screen for enzymes involved in plant biomass degradation are based on the use of single substrates. Moreover, highly diverse environments are used as metagenomic sources. However, such methods suffer from low hit rates of positive clones and hence the discovery of novel enzymatic activities from metagenomes has been hampered. Here, we constructed fosmid libraries from two wheat straw-degrading microbial consortia, denoted RWS (bred on untreated wheat straw) and TWS (bred on heat-treated wheat straw). Approximately 22,000 clones from each library were screened for (hemi)cellulose-degrading enzymes using a multi-chromogenic substrate approach. The screens yielded 71 positive clones for both libraries, giving hit rates of 1:440 and 1:1,047 for RWS and TWS, respectively. Seven clones (NT2-2, T5-5, NT18-17, T4-1, 10BT, NT18-21 and T17-2) were selected for sequence analyses. Their inserts revealed the presence of 18 genes encoding enzymes belonging to twelve different glycosyl hydrolase families (GH2, GH3, GH13, GH17, GH20, GH27, GH32, GH39, GH53, GH58, GH65 and GH109). These encompassed several carbohydrate-active gene clusters traceable mainly to Klebsiella related species. Detailed functional analyses showed that clone NT2-2 (containing a beta-galactosidase of ~116 kDa) had highest enzymatic activity at 55 °C and pH 9.0. Additionally, clone T5-5 (containing a beta-xylosidase of ~86 kDa) showed > 90% of enzymatic activity at 55 °C and pH 10.0. This study employed a high-throughput method for rapid screening of fosmid metagenomic libraries for (hemi)cellulose-degrading enzymes. The approach, consisting of screens on multi-substrates coupled to further analyses, revealed high hit rates, as compared with recent other studies. Two

  11. Post-fire drought effects and their legacy on soil functionality and microbial community structure in a Mediterranean shrubland

    Science.gov (United States)

    Belen Hinojosa, M.; Parra, Antonio; Laudicina, V. Armando; Moreno, José M.

    2017-04-01

    Climate change in subtropical areas, like the Mediterranean, is projected to decrease precipitation and to lengthen the seasonal drought period. Fire danger is also projected to increase under the most severe conditions. Little is known about the effects of increasing drought and, particularly, its legacy when precipitation resumes to normal, on the recovery of burned ecosystems. Here we studied the effects of post-fire drought and its legacy two years after it stopped on soil microbial community structure and functionality of a Cistus-Erica shrubland. To do this, a manipulative experiment was setup in which rainfall total patterns were modified by means of a rain-out shelters and irrigation system in a fully replicated set of previously burned plots. The treatments were: environmental control (natural rainfall), historical control (average rainfall, 2 months drought), moderate drought (25% reduction of historical control, 5 months drought) and severe drought (45% reduction, 7 months drought). One set of unburned plots under natural rainfall served as an additional control. Availability of the main soil nutrients and microbial community composition and functionality were monitored over 4 years under these rainfall manipulation treatments. Thereafter, treatments were discontinued and plots were subjected to ambient rainfall for two additional years. Post-fire drought had not effect on total C or N. Fire increased soil P and N availability. However, post-fire drought reduced available soil P and increased nitrate in the short term. Post- fire reduction of available K was accentuated by continued drought. Fire significantly reduced soil organic matter, enzyme activities and carbon mineralization, mainly in drought treated soils. Fire also decreased soil microbial biomass and the proportion of fungi, while that of actinomycetes increased in the short term. Post-fire drought accentuated the decrease of soil total microbial biomass and fungi, with bacteria becoming more

  12. Laboratory-scale method for enzymatic saccharification of lignocellulosic biomass at high-solids loadings

    Directory of Open Access Journals (Sweden)

    Dibble Clare J

    2009-11-01

    Full Text Available Abstract Background Screening new lignocellulosic biomass pretreatments and advanced enzyme systems at process relevant conditions is a key factor in the development of economically viable lignocellulosic ethanol. Shake flasks, the reaction vessel commonly used for screening enzymatic saccharifications of cellulosic biomass, do not provide adequate mixing at high-solids concentrations when shaking is not supplemented with hand mixing. Results We identified roller bottle reactors (RBRs as laboratory-scale reaction vessels that can provide adequate mixing for enzymatic saccharifications at high-solids biomass loadings without any additional hand mixing. Using the RBRs, we developed a method for screening both pretreated biomass and enzyme systems at process-relevant conditions. RBRs were shown to be scalable between 125 mL and 2 L. Results from enzymatic saccharifications of five biomass pretreatments of different severities and two enzyme preparations suggest that this system will work well for a variety of biomass substrates and enzyme systems. A study of intermittent mixing regimes suggests that mass transfer limitations of enzymatic saccharifications at high-solids loadings are significant but can be mitigated with a relatively low amount of mixing input. Conclusion Effective initial mixing to promote good enzyme distribution and continued, but not necessarily continuous, mixing is necessary in order to facilitate high biomass conversion rates. The simplicity and robustness of the bench-scale RBR system, combined with its ability to accommodate numerous reaction vessels, will be useful in screening new biomass pretreatments and advanced enzyme systems at high-solids loadings.

  13. Shift in soil microbial communities with shrub encroachment in Inner Mongolia grasslands, China

    Science.gov (United States)

    Shen, H.; Li, H.; Zhang, J.; Hu, H.; Chen, L.; Zhu, Y.; Fang, J.

    2017-12-01

    The ongoing expansion of shrub encroachment into grasslands represents a unique form of land cover change. How this process affects soil microbial communities is poorly understood. In this study, we aim to assess the effects of shrub encroachment on soil microbial biomass, abundance and composition by comparing data between shrub patches and neighboring herb patches in shrub-encroached grasslands (SEGs) in Inner Mongolia, China. Fourteen SEG sites from two ecosystem types (typical and desert grasslands) were investigated. The phospholipid fatty acid (PLFA) method was used to analyze the composition and biomass of the soil microbial community. Our results showed that the top-soil microbial biomass and abundances of gram-negative bacteria, arbuscular mycorrhizal fungi, and actinomycetes were significantly higher in shrub patches than in herb patches in both typical and desert grasslands (P fungi to bacteria ratio was significantly higher in shrub patches than in herb patches in desert grassland (P soil microbial communities, which makes the microbial communities toward a fresh organic carbon-based structure. This study highlights the importance of edaphic and climate factors in microbial community shifts in SEGs.

  14. Microbial biomass in compost during colonization of Agaricus bisporus

    NARCIS (Netherlands)

    Vos, Aurin M.; Heijboer, Amber; Boschker, Henricus T.S.; Bonnet, Barbara; Lugones, Luis G.; Wösten, Han A.B.

    2017-01-01

    Agaricus bisporus mushrooms are commercially produced on a microbe rich compost. Here, fungal and bacterial biomass was quantified in compost with and without colonization by A. bisporus. Chitin content, indicative of total fungal biomass, increased during a 26-day period from 576 to 779 nmol

  15. Generation of Electricity and Analysis of Microbial Communities in Wheat Straw Biomass-Powered Microbial Fuel Cells

    DEFF Research Database (Denmark)

    Zhang, Yifeng; Min, Booki; Huang, L.

    2009-01-01

    Electricity generation from wheat straw hydrolysate and the microbial ecology of electricity producing microbial communities developed in two chamber microbial fuel cells (MFCs) were investigated. Power density reached 123 mW/m2 with an initial hydrolysate concentration of 1000 mg-COD/L while...

  16. Comparison of seasonal soil microbial process in snow-covered temperate ecosystems of northern China.

    Directory of Open Access Journals (Sweden)

    Xinyue Zhang

    Full Text Available More than half of the earth's terrestrial surface currently experiences seasonal snow cover and soil frost. Winter compositional and functional investigations in soil microbial community are frequently conducted in alpine tundra and boreal forest ecosystems. However, little information on winter microbial biogeochemistry is known from seasonally snow-covered temperate ecosystems. As decomposer microbes may differ in their ability/strategy to efficiently use soil organic carbon (SOC within different phases of the year, understanding seasonal microbial process will increase our knowledge of biogeochemical cycling from the aspect of decomposition rates and corresponding nutrient dynamics. In this study, we measured soil microbial biomass, community composition and potential SOC mineralization rates in winter and summer, from six temperate ecosystems in northern China. Our results showed a clear pattern of increased microbial biomass C to nitrogen (N ratio in most winter soils. Concurrently, a shift in soil microbial community composition occurred with higher fungal to bacterial biomass ratio and gram negative (G- to gram positive (G+ bacterial biomass ratio in winter than in summer. Furthermore, potential SOC mineralization rate was higher in winter than in summer. Our study demonstrated a distinct transition of microbial community structure and function from winter to summer in temperate snow-covered ecosystems. Microbial N immobilization in winter may not be the major contributor for plant growth in the following spring.

  17. Engineering Cellulase Enzymes for Bioenergy

    Science.gov (United States)

    Atreya, Meera Elizabeth

    Sustainable energy sources, such as biofuels, offer increasingly important alternatives to fossil fuels that contribute less to global climate change. The energy contained within cellulosic biofuels derives from sunlight energy stored in the form of carbon-carbon bonds comprising sugars such as glucose. Second-generation biofuels are produced from lignocellulosic biomass feedstocks, including agricultural waste products and non-food crops like Miscanthus, that contain lignin and the polysaccharides hemicellulose and cellulose. Cellulose is the most abundant biological material on Earth; it is a polymer of glucose and a structural component of plant cell walls. Accessing the sugar is challenging, as the crystalline structure of cellulose resists degradation; biochemical and thermochemical means can be used to depolymerize cellulose. Cellulase enzymes catalyze the biochemical depolymerization of cellulose into glucose. Glucose can be used as a carbon source for growth of a biofuel-producing microorganism. When it converts glucose to a hydrocarbon fuel, this microbe completes the biofuels process of transforming sunlight energy into accessible, chemical energy capable of replacing non-renewable transportation fuels. Due to strong intermolecular interactions between polymer chains, cellulose is significantly more challenging to depolymerize than starch, a more accessible polymer of glucose utilized in first-generation biofuels processes (often derived from corn). While most mammals cannot digest cellulose (dietary fiber), certain fungi and bacteria produce cellulase enzymes capable of hydrolyzing it. These organisms secrete a wide variety of glycoside hydrolase and other classes of enzymes that work in concert. Because cellulase enzymes are slow-acting and expensive to produce, my aim has been to improve the properties of these enzymes as a means to make a cellulosic biofuels process possible that is more efficient and, consequently, more economical than current

  18. Microbial growth and substrate utilization kinetics | Okpokwasili ...

    African Journals Online (AJOL)

    Microbial growth on and utilization of environmental contaminants as substrates have been studied by many researchers. Most times, substrate utilization results in removal of chemical contaminant, increase in microbial biomass and subsequent biodegradation of the contaminant. These are all aimed at detoxification of the ...

  19. Spatial variability of enzyme activities and microbial biomass in the upper layers of Quercus petraea forest soil

    Czech Academy of Sciences Publication Activity Database

    Šnajdr, Jaroslav; Valášková, Vendula; Merhautová, Věra; Herinková, Jana; Cajthaml, Tomáš; Baldrian, Petr

    2008-01-01

    Roč. 40, č. 9 (2008), s. 2068-2075 ISSN 0038-0717 R&D Projects: GA MŠk LC06066; GA MZe QH72216; GA AV ČR KJB600200516 Institutional research plan: CEZ:AV0Z50200510 Keywords : enzyme activity * forest soil * lignocellulose Subject RIV: EE - Microbiology, Virology Impact factor: 2.926, year: 2008

  20. Mineralogic control on abundance and diversity of surface-adherent microbial communities

    Science.gov (United States)

    Mauck, Brena S.; Roberts, Jennifer A.

    2007-01-01

    In this study, we investigated the role of mineral-bound P and Fe in defining microbial abundance and diversity in a carbon-rich groundwater. Field colonization experiments of initially sterile mineral surfaces were combined with community structure characterization of the attached microbial population. Silicate minerals containing varying concentrations of P (∼1000 ppm P) and Fe (∼4 wt % Fe 2 O3), goethite (FeOOH), and apatite [Ca5(PO4)3(OH)] were incubated for 14 months in three biogeochemically distinct zones within a petroleum-contaminated aquifer. Phospholipid fatty acid analysis of incubated mineral surfaces and groundwater was used as a measure of microbial community structure and biomass. Microbial biomass on minerals exhibited distinct trends as a function of mineralogy depending on the environment of incubation. In the carbon-rich, aerobic groundwater attached biomass did not correlate to the P- or Fe- content of the mineral. In the methanogenic groundwater, however, biomass was most abundant on P-containing minerals. Similarly, in the Fe-reducing groundwater a correlation between Fe-content and biomass was observed. The community structure of the mineral-adherent microbial population was compared to the native groundwater community. These two populations were significantly different regardless of mineralogy, suggesting differentiation of the planktonic community through attachment, growth, and death of colonizing cells. Biomarkers specific for dissimilatory Fe-reducing bacteria native to the aquifer were identified only on Fe-containing minerals in the Fe-reducing groundwater. These results demonstrate that the trace nutrient content of minerals affects both the abundance and diversity of surface-adherent microbial communities. This behavior may be a means to access limiting nutrients from the mineral, creating a niche for a particular microbial population. These results suggest that heterogeneity of microbial populations and their associated

  1. Impact of Transgenic Brassica napus Harboring the Antifungal Synthetic Chitinase (NiC Gene on Rhizosphere Microbial Diversity and Enzyme Activities

    Directory of Open Access Journals (Sweden)

    Mohammad S. Khan

    2017-07-01

    Full Text Available Transgenic Brassica napus harboring the synthetic chitinase (NiC gene exhibits broad-spectrum antifungal resistance. As the rhizosphere microorganisms play an important role in element cycling and nutrient transformation, therefore, biosafety assessment of NiC containing transgenic plants on soil ecosystem is a regulatory requirement. The current study is designed to evaluate the impact of NiC gene on the rhizosphere enzyme activities and microbial community structure. The transgenic lines with the synthetic chitinase gene (NiC showed resistance to Alternaria brassicicola, a common disease causing fungal pathogen. The rhizosphere enzyme analysis showed no significant difference in the activities of fivesoil enzymes: alkalyine phosphomonoestarase, arylsulphatase, β-glucosidase, urease and sucrase between the transgenic and non-transgenic lines of B. napus varieties, Durr-e-NIFA (DN and Abasyne-95 (AB-95. However, varietal differences were observed based on the analysis of molecular variance. Some individual enzymes were significantly different in the transgenic lines from those of non-transgenic but the results were not reproducible in the second trail and thus were considered as environmental effect. Genotypic diversity of soil microbes through 16S–23S rRNA intergenic spacer region amplification was conducted to evaluate the potential impact of the transgene. No significant diversity (4% for bacteria and 12% for fungal between soil microbes of NiC B. napus and the non-transgenic lines was found. However, significant varietal differences were observed between DN and AB-95 with 79% for bacterial and 54% for fungal diversity. We conclude that the NiC B. napus lines may not affect the microbial enzyme activities and community structure of the rhizosphere soil. Varietal differences might be responsible for minor changes in the tested parameters.

  2. Inclusion of caraway in the ryegrass-red clover mixture modifies soil microbial community composition

    DEFF Research Database (Denmark)

    Cong, Wenfeng; Jing, Jingying; Søegaard, Karen

    -containing grass-clover mixtures may potentially affect soil microbial community structure, biomass and associated ecosystem functions, but it is yet to be elucidated. We hypothesized that inclusion of plantain in the grass-clover mixture would enhance soil microbial biomas and functions through its high biomass...

  3. Soil microbial biomass under pine forests in the north-western Spain: influence of stand age, site index and parent material

    Energy Technology Data Exchange (ETDEWEB)

    Mahia, J.; Perez-Ventura, L.; Cabaneiro, A.; Diaz-Ravina, M.

    2006-07-01

    The effects of stand age, site index and parent material on soil biochemical properties related to biomass (extractable C, microbial C and metabolic quotient) were examined in the 0-15 cm mineral soil layers of Pinus pinaster and Pinus sylvestris stand from NW Spain. Two productivity levels (low and high site index), two ages (young and old) and two parent soil materials (granite and acid schists) were considered. The data indicated that there were differences in microbial parameters in soils under different species. In general in P. pinaster forest higher values of biochemical parameters expressed on organic C basis, were observed in the stands of high site index as compared with the low ones; in contrast, in P. sylvestris no differences among stand site index were detected. In both species different results were also observed depending on parent material and a significant effect of stand age was detected for extractable C and microbial C in P. pinaster forest developed over granite. The data seem to indicate that measured parameters may have the potential to be used as indicators of the effect of forest management on soil organic matter quality. (Author) 25 refs.

  4. Are variations in heterotrophic soil respiration related to changes in substrate availability and microbial biomass carbon in the subtropical forests?

    Science.gov (United States)

    Wei, Hui; Chen, Xiaomei; Xiao, Guoliang; Guenet, Bertrand; Vicca, Sara; Shen, Weijun

    2015-12-16

    Soil temperature and moisture are widely-recognized controlling factors on heterotrophic soil respiration (Rh), although they often explain only a portion of Rh variability. How other soil physicochemical and microbial properties may contribute to Rh variability has been less studied. We conducted field measurements on Rh half-monthly and associated soil properties monthly for two years in four subtropical forests of southern China to assess influences of carbon availability and microbial properties on Rh. Rh in coniferous forest was significantly lower than that in the other three broadleaf species-dominated forests and exhibited obvious seasonal variations in the four forests (P forests. The quantity and decomposability of dissolved organic carbon (DOC) were significantly important to Rh variations, but the effect of DOC content on Rh was confounded with temperature, as revealed by partial mantel test. Microbial biomass carbon (MBC) was significantly related to Rh variations across forests during the warm season (P = 0.043). Our results suggest that DOC and MBC may be important when predicting Rh under some conditions, and highlight the complexity by mutual effects of them with environmental factors on Rh variations.

  5. Novel optimization strategy for tannase production through a modified solid-state fermentation system.

    Science.gov (United States)

    Wu, Changzheng; Zhang, Feng; Li, Lijun; Jiang, Zhedong; Ni, Hui; Xiao, Anfeng

    2018-01-01

    High amounts of insoluble substrates exist in the traditional solid-state fermentation (SSF) system. The presence of these substrates complicates the determination of microbial biomass. Thus, enzyme activity is used as the sole index for the optimization of the traditional SSF system, and the relationship between microbial growth and enzyme synthesis is always ignored. This study was conducted to address this deficiency. All soluble nutrients from tea stalk were extracted using water. The aqueous extract was then mixed with polyurethane sponge to establish a modified SSF system, which was then used to conduct tannase production. With this system, biomass, enzyme activity, and enzyme productivity could be measured rationally and accurately. Thus, the association between biomass and enzyme activity could be easily identified, and the shortcomings of traditional SSF could be addressed. Different carbon and nitrogen sources exerted different effects on microbial growth and enzyme production. Single-factor experiments showed that glucose and yeast extract greatly improved microbial biomass accumulation and that tannin and (NH 4 ) 2 SO 4 efficiently promoted enzyme productivity. Then, these four factors were optimized through response surface methodology. Tannase activity reached 19.22 U/gds when the added amounts of tannin, glucose, (NH 4 ) 2 SO 4 , and yeast extract were 7.49, 8.11, 9.26, and 2.25%, respectively. Tannase activity under the optimized process conditions was 6.36 times higher than that under the initial process conditions. The optimized parameters were directly applied to the traditional tea stalk SSF system. Tannase activity reached 245 U/gds, which is 2.9 times higher than our previously reported value. In this study, a modified SSF system was established to address the shortcomings of the traditional SSF system. Analysis revealed that enzymatic activity and microbial biomass are closely related, and different carbon and nitrogen sources have different

  6. Biological strategies for enhanced hydrolysis of lignocellulosic biomass during anaerobic digestion: Current status and future perspectives.

    Science.gov (United States)

    Shrestha, Shilva; Fonoll, Xavier; Khanal, Samir Kumar; Raskin, Lutgarde

    2017-12-01

    Lignocellulosic biomass is the most abundant renewable bioresource on earth. In lignocellulosic biomass, the cellulose and hemicellulose are bound with lignin and other molecules to form a complex structure not easily accessible to microbial degradation. Anaerobic digestion (AD) of lignocellulosic biomass with a focus on improving hydrolysis, the rate limiting step in AD of lignocellulosic feedstocks, has received considerable attention. This review highlights challenges with AD of lignocellulosic biomass, factors contributing to its recalcitrance, and natural microbial ecosystems, such as the gastrointestinal tracts of herbivorous animals, capable of performing hydrolysis efficiently. Biological strategies that have been evaluated to enhance hydrolysis of lignocellulosic biomass include biological pretreatment, co-digestion, and inoculum selection. Strategies to further improve these approaches along with future research directions are outlined with a focus on linking studies of microbial communities involved in hydrolysis of lignocellulosics to process engineering. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. Compounds Released from Biomass Deconstruction: Understanding Their Effect on Cellulose Enzyme Hydrolysis and Their Biological Activity

    Science.gov (United States)

    Djioleu, Angele Mezindjou

    The effect of compounds produced during biomass pretreatment on cellulolytic enzyme was investigated. Liquid prehydrolyzates were prepared by pretreating switchgrass using 24 combinations of temperature, time, and sulfuric acid concentration based on a full factorial design. Temperature was varied from 140°C to 180°C; time ranged from 10 to 40 min; and the sulfuric acid concentrations were 0.5% or 1% (v/v). Identified products in the prehydrolyzates included xylose, glucose, hydroxymethylfurfural (HMF), furfural, acetic acid, formic acid, and phenolic compounds at concentration ranging from 0 to 21.4 g/L. Pretreatment conditions significantly affected the concentrations of compounds detected in prehydrolyzates. When assayed in the presence of switchgrass prehydrolyzates against model substrates, activities of cellulase, betaglucosidase, and exoglucanase, were significantly reduced by at least 16%, 31.8%, and 57.8%, respectively, as compared to the control. A strong positive correlation between inhibition of betaglucosidase and concentration of glucose, acetic acid, and furans in prehydrolyzate was established. Exoglucanase inhibition correlated with the presence of phenolic compounds and acetic acid. The prehydrolyzate, prepared at 160°C, 30 min, and 1% acid, was fractionated by centrifugal partition chromatography (CPC) into six fractions; the inhibition effect of these fractions on betaglucosidase and exoglucanase was determined. The initial hydrolysis rate of cellobiose by betaglucosidase was significantly reduced by the CPC sugar-rich fraction; however, exoglucanase was deactivated by the CPC phenolic-rich fraction. Finally, biological activities of water-extracted compounds from sweetgum bark and their effect on cellulase was investigated. It was determined that 12% of solid content of the bark extract could be accounted by phenolic compounds with gallic acid identified as the most concentrated phytochemical. Sweetgum bark extract inhibited Staphylococcus

  8. Assessment of Cu applications in two contrasting soils-effects on soil microbial activity and the fungal community structure.

    Science.gov (United States)

    Keiblinger, Katharina M; Schneider, Martin; Gorfer, Markus; Paumann, Melanie; Deltedesco, Evi; Berger, Harald; Jöchlinger, Lisa; Mentler, Axel; Zechmeister-Boltenstern, Sophie; Soja, Gerhard; Zehetner, Franz

    2018-03-01

    Copper (Cu)-based fungicides have been used in viticulture to prevent downy mildew since the end of the 19th century, and are still used today to reduce fungal diseases. Consequently, Cu has built up in many vineyard soils, and it is still unclear how this affects soil functioning. The present study aimed to assess the short and medium-term effects of Cu contamination on the soil fungal community. Two contrasting agricultural soils, an acidic sandy loam and an alkaline silt loam, were used for an eco-toxicological greenhouse pot experiment. The soils were spiked with a Cu-based fungicide in seven concentrations (0-5000 mg Cu kg -1 soil) and alfalfa was grown in the pots for 3 months. Sampling was conducted at the beginning and at the end of the study period to test Cu toxicity effects on total microbial biomass, basal respiration and enzyme activities. Fungal abundance was analysed by ergosterol at both samplings, and for the second sampling, fungal community structure was evaluated via ITS amplicon sequences. Soil microbial biomass C as well as microbial respiration rate decreased with increasing Cu concentrations, with EC 50 ranging from 76 to 187 mg EDTA-extractable Cu kg -1 soil. Oxidative enzymes showed a trend of increasing activity at the first sampling, but a decline in peroxidase activity was observed for the second sampling. We found remarkable Cu-induced changes in fungal community abundance (EC 50 ranging from 9.2 to 94 mg EDTA-extractable Cu kg -1 soil) and composition, but not in diversity. A large number of diverse fungi were able to thrive under elevated Cu concentrations, though within the order of Hypocreales several species declined. A remarkable Cu-induced change in the community composition was found, which depended on the soil properties and, hence, on Cu availability.

  9. Influence of Inoculation, Nitrogen and Phosphorus Levels on Wheat Growth and Soil Microbial Biomass-N Using 15N Techniques

    International Nuclear Information System (INIS)

    Galal, Y.G.; El-Ghandour, I.A.; Abdel Raouf, A.M.; Osman, M.E.

    2003-01-01

    Pot experiment was carried out with wheat that cultivated in virgin sandy soil and inoculated with Rhizobium (Rh), mycorrhizea (VAM) and mixture of both. The objective of this work was to verify the potential of these inoculum on wheat production, nutrient acquisition and microbial biomass N (MBN) contribution as affected by N and P fertilizers levels. MBN was detected through the fumigation-extraction method. Nitrogen and phosphorus fertilizers were applied at three levels, 0; 25 ppm N and 3.3 ppm P and 50 ppm N and 6.6 ppm P in the form of ( 15 NH 4 ) 2 SO 4 , 5% atom excess and super-phosphate, respectively. The effect of inoculation and chemical fertilizers on dry matter (DM), N and P uptake (shoot and grain) and MBN were traced. The obtained data revealed that the highest DM and N uptake by wheat shoot were recorded with the dual inoculation (Rh + VAM) at the highest level of N and P fertilizers. The highest grain yield was detected with single inoculum of AM fungi while N and P uptake were with dual inoculation at the same rate of fertilizers. Inoculation with Rh either alone or in combination with VAM have a positive and stimulative effect on wheat growth and N and P uptake indicating the possibilities of extending the use of symbiotic microorganisms to be applied with cereals. The fluctuation in the soil microbial biomass N did not gave a chance to recognize, exactly, the impact of inoculation and/or fertilization levels

  10. Something from (almost) nothing: the impact of multiple displacement amplification on microbial ecology.

    Science.gov (United States)

    Binga, Erik K; Lasken, Roger S; Neufeld, Josh D

    2008-03-01

    Microbial ecology is a field that applies molecular techniques to analyze genes and communities associated with a plethora of unique environments on this planet. In the past, low biomass and the predominance of a few abundant community members have impeded the application of techniques such as PCR, microarray analysis and metagenomics to complex microbial populations. In the absence of suitable cultivation methods, it was not possible to obtain DNA samples from individual microorganisms. Recently, a method called multiple displacement amplification (MDA) has been used to circumvent these limitations by amplifying DNA from microbial communities in low-biomass environments, individual cells from uncultivated microbial species and active organisms obtained through stable isotope probing incubations. This review describes the development and applications of MDA, discusses its strengths and limitations and highlights the impact of MDA on the field of microbial ecology. Whole genome amplification via MDA has increased access to the genomic DNA of uncultivated microorganisms and low-biomass environments and represents a 'power tool' in the molecular toolbox of microbial ecologists.

  11. Investigating commercial cellulase performances toward specific biomass recalcitrance factors using reference substrates.

    Science.gov (United States)

    Ju, Xiaohui; Bowden, Mark; Engelhard, Mark; Zhang, Xiao

    2014-05-01

    Three commercial cellulase preparations, Novozymes Cellic(®) Ctec2, Dupont Accellerase(®) 1500, and DSM Cytolase CL, were evaluated for their hydrolytic activity using a set of reference biomass substrates with controlled substrate characteristics. It was found that lignin remains a significant recalcitrance factor to all the preparations, although different enzyme preparations respond to the inhibitory effect of lignin differently. Also, different types of biomass lignin can inhibit cellulase enzymes in different manners. Enhancing enzyme activity toward biomass fiber swelling is an area significantly contributing to potential improvement in cellulase performance. While the degree of polymerization of cellulose in the reference substrates did not present a major recalcitrance factor to Novozymes Cellic(®) Ctec2, cellulose crystallite has been shown to have a significant lower reactivity toward all enzyme mixtures. The presence of polysaccharide monooxygenases (PMOs) in Novozymes Ctec2 appears to enhance enzyme activity toward decrystallization of cellulose. This study demonstrated that reference substrates with controlled chemical and physical characteristics of structural features can be applied as an effective and practical strategy to identify cellulosic enzyme activities toward specific biomass recalcitrance factor(s) and provide specific targets for enzyme improvement.

  12. Biotechnological Processes in Microbial Amylase Production.

    Science.gov (United States)

    Gopinath, Subash C B; Anbu, Periasamy; Arshad, M K Md; Lakshmipriya, Thangavel; Voon, Chun Hong; Hashim, Uda; Chinni, Suresh V

    2017-01-01

    Amylase is an important and indispensable enzyme that plays a pivotal role in the field of biotechnology. It is produced mainly from microbial sources and is used in many industries. Industrial sectors with top-down and bottom-up approaches are currently focusing on improving microbial amylase production levels by implementing bioengineering technologies. The further support of energy consumption studies, such as those on thermodynamics, pinch technology, and environment-friendly technologies, has hastened the large-scale production of the enzyme. Herein, the importance of microbial (bacteria and fungi) amylase is discussed along with its production methods from the laboratory to industrial scales.

  13. A novel feruloyl esterase from rumen microbial metagenome: Gene cloning and enzyme characterization in the release of mono- and diferulic acids

    Science.gov (United States)

    A feruloyl esterase (FAE) gene was isolated from a rumen microbial metagenome, cloned into E. coli, and expressed in active form. The enzyme (RuFae4) was classified as a Type D feruloyl esterase based on its action on synthetic substrates and ability to release diferulates. The RuFae4 alone releas...

  14. Exploration of soil metagenome diversity for prospection of enzymes involved in lignocellulosic biomass conversion

    Energy Technology Data Exchange (ETDEWEB)

    Alvarez, T.M.; Squina, F.M. [Laboratorio Nacional de Luz Sincrotron (LNLS), Campinas, SP (Brazil); Paixao, D.A.A.; Franco Cairo, J.P.L.; Buchli, F.; Ruller, R. [Laboratorio Nacional de Ciencia e Tecnologia do Bioetanol (CTBE), Campinas, SP (Brazil); Prade, R. [Oklahoma State University, Sillwater, OK (United States)

    2012-07-01

    Full text: Metagenomics allows access to genetic information encoded in DNA of microorganisms recalcitrant to cultivation. They represent a reservoir of novel biocatalyst with potential application in environmental friendly techniques aiming to overcome the dependence on fossil fuels and also to diminish air and water pollution. The focus of our work is the generation of a tool kit of lignocellulolytic enzymes from soil metagenome, which could be used for second generation ethanol production. Environmental samples were collected at a sugarcane field after harvesting, where it is expected that the microbial population involved on lignocellulose degradation was enriched due to the presence of straws covering the soil. Sugarcane Bagasse-Degrading-Soil (SBDS) metagenome was massively-parallel-454-Roche-sequenced. We identified a full repertoire of genes with significant match to glycosyl hydrolases catalytic domain and carbohydrate-binding modules. Soil metagenomics libraries cloned into pUC19 were screened through functional assays. CMC-agar screening resulted in positive clones, revealing new cellulases coding genes. Through a CMC-zymogram it was possible to observe that one of these genes, nominated as E-1, corresponds to an enzyme that is secreted to the extracellular medium, suggesting that the cloned gene carried the original signal peptide. Enzymatic assays and analysis through capillary electrophoresis showed that E-1 was able to cleave internal glycosidic bonds of cellulose. New rounds of functional screenings through chromogenic substrates are being conducted aiming the generation of a library of lignocellulolytic enzymes derived from soil metagenome, which may become key component for development of second generation biofuels. (author)

  15. Microbial activity in soil cultivated with different summer legumes in coffee crop

    Directory of Open Access Journals (Sweden)

    Elcio Liborio Balota

    2011-02-01

    Full Text Available A field experiment was conducted for ten years in a sandy soil in the north part of the Paraná State, Brazil. The soil samples were collected at 0-10 cm depth, both under the coffee canopy and in the inter row space between the coffee plants, in the following treatments: Control, Leucaena leucocephala, Crotalaria spectabilis, Crotalaria breviflora, Mucuna pruriens, Mucuna deeringiana, Arachis hypogaea and Vigna unguiculata. The legume crops influenced the microbial activity, both under the coffee canopy and in the inter row space. The cultivation of Leucaena leucocephala increased the microbial biomass C, N and P. Although L. leucocephala and Arachis hypogaea provided higher microbial biomass, the qCO2 decreased by up to 50% under the coffee canopy and by about 25% in the inter row space. The soil microbial biomass was enriched in N and P due to green manure residue addition.

  16. Nitrogen fertilization decouples roots and microbes: Reductions in belowground carbon allocation limit microbial activity

    Science.gov (United States)

    Carrara, J.; Walter, C. A.; Govindarajulu, R.; Hawkins, J.; Brzostek, E. R.

    2017-12-01

    Nitrogen (N) deposition has enhanced the ability of trees to capture atmospheric carbon (C). The effect of elevated N on belowground C cycling, however, is variable and response mechanisms are largely unknown. Recent research has highlighted distinct differences between ectomycorrhizal (ECM) and arbuscular mycorrhizal (AM) trees in the strength of root-microbial interactions. In particular, ECM trees send more C to rhizosphere microbes to stimulate enzyme activity and nutrient mobilization than AM trees, which primarily rely on saprotrophic microbes to mobilize N. As such, we hypothesized that N fertilization would weaken root-microbial interactions and soil decomposition in ECM stands more than in AM stands. To test this hypothesis, we measured root-microbial interactions in ECM and AM plots in two long-term N fertilization studies, the Fernow Experimental Forest, WV and Bear Brook Watershed, ME. We found that N fertilization led to declines in plant C allocation belowground to fine root biomass, branching, and root exudation in ECM stands to a greater extent than in AM stands. As ECM roots are tightly coupled to the soil microbiome through energy and nutrient exchange, reductions in belowground C allocation were mirrored by shifts in microbial community composition and reductions in fungal gene expression. These shifts were accompanied by larger reductions in fungal-derived lignolytic and hydrolytic enzyme activity in ECM stands than in AM stands. In contrast, as the AM soil microbiome is less reliant on trees for C and are more adapted to high inorganic nutrient environments, the soil metagenome and transcriptome were more resilient to decreases in belowground C allocation. Collectively, our results indicate the N fertilization decoupled root-microbial interactions by reducing belowground carbon allocation in ECM stands. Thus, N fertilization may reduce soil turnover and increase soil C storage to a greater extent in forests dominated by ECM than AM trees.

  17. Recent patents on genetic modification of plants and microbes for biomass conversion to biofuels.

    Science.gov (United States)

    Lubieniechi, Simona; Peranantham, Thinesh; Levin, David B

    2013-04-01

    Development of sustainable energy systems based on renewable biomass feedstocks is now a global effort. Lignocellulosic biomass contains polymers of cellulose, hemicellulose, and lignin, bound together in a complex structure. Liquid biofuels, such as ethanol, can be made from biomass via fermentation of sugars derived from the cellulose and hemicellulose within lignocellulosic materials, but pre-treatment of the biomass to release sugars for microbial conversion is a significant barrier to commercial success of lignocellulosic biofuel production. Strategies to reduce the energy and cost inputs required for biomass pre-treatment include genetic modification of plant materials to reduce lignin content. Significant efforts are also underway to create recombinant microorganisms capable of converting sugars derived from lignocellulosic biomass to a variety of biofuels. An alternative strategy to reduce the costs of cellulosic biofuel production is the use of cellulolytic microorganisms capable of direct microbial conversion of ligno-cellulosic biomass to fuels. This paper reviews recent patents on genetic modification of plants and microbes for biomass conversion to biofuels.

  18. Susceptibility of green and conventional building materials to microbial growth.

    Science.gov (United States)

    Mensah-Attipoe, J; Reponen, T; Salmela, A; Veijalainen, A-M; Pasanen, P

    2015-06-01

    Green building materials are becoming more popular. However, little is known about their ability to support or limit microbial growth. The growth of fungi was evaluated on five building materials. Two green, two conventional building materials and wood as a positive control were selected. The materials were inoculated with Aspergillus versicolor, Cladosporium cladosporioides and Penicillium brevicompactum, in the absence and presence of house dust. Microbial growth was assessed at four different time points by cultivation and determining fungal biomass using the N-acetylhexosaminidase (NAHA) enzyme assay. No clear differences were seen between green and conventional building materials in their susceptibility to support microbial growth. The presence of dust, an external source of nutrients, promoted growth of all the fungal species similarly on green and conventional materials. The results also showed a correlation coefficient ranging from 0.81 to 0.88 between NAHA activity and culturable counts. The results suggest that the growth of microbes on a material surface depends on the availability of organic matter rather than the classification of the material as green or conventional. NAHA activity and culturability correlated well indicating that the two methods used in the experiments gave similar trends for the growth of fungi on material surfaces. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  19. Genome-enabled Modeling of Microbial Biogeochemistry using a Trait-based Approach. Does Increasing Metabolic Complexity Increase Predictive Capabilities?

    Science.gov (United States)

    King, E.; Karaoz, U.; Molins, S.; Bouskill, N.; Anantharaman, K.; Beller, H. R.; Banfield, J. F.; Steefel, C. I.; Brodie, E.

    2015-12-01

    The biogeochemical functioning of ecosystems is shaped in part by genomic information stored in the subsurface microbiome. Cultivation-independent approaches allow us to extract this information through reconstruction of thousands of genomes from a microbial community. Analysis of these genomes, in turn, gives an indication of the organisms present and their functional roles. However, metagenomic analyses can currently deliver thousands of different genomes that range in abundance/importance, requiring the identification and assimilation of key physiologies and metabolisms to be represented as traits for successful simulation of subsurface processes. Here we focus on incorporating -omics information into BioCrunch, a genome-informed trait-based model that represents the diversity of microbial functional processes within a reactive transport framework. This approach models the rate of nutrient uptake and the thermodynamics of coupled electron donors and acceptors for a range of microbial metabolisms including heterotrophs and chemolithotrophs. Metabolism of exogenous substrates fuels catabolic and anabolic processes, with the proportion of energy used for cellular maintenance, respiration, biomass development, and enzyme production based upon dynamic intracellular and environmental conditions. This internal resource partitioning represents a trade-off against biomass formation and results in microbial community emergence across a fitness landscape. Biocrunch was used here in simulations that included organisms and metabolic pathways derived from a dataset of ~1200 non-redundant genomes reflecting a microbial community in a floodplain aquifer. Metagenomic data was directly used to parameterize trait values related to growth and to identify trait linkages associated with respiration, fermentation, and key enzymatic functions such as plant polymer degradation. Simulations spanned a range of metabolic complexities and highlight benefits originating from simulations

  20. The nitrogen cycle in anaerobic methanotrophic mats of the Black Sea is linked to sulfate reduction and biomass decomposition.

    Science.gov (United States)

    Siegert, Michael; Taubert, Martin; Seifert, Jana; von Bergen-Tomm, Martin; Basen, Mirko; Bastida, Felipe; Gehre, Matthias; Richnow, Hans-Hermann; Krüger, Martin

    2013-11-01

    Anaerobic methanotrophic (ANME) mats host methane-oxidizing archaea and sulfate-reducing prokaryotes. Little is known about the nitrogen cycle in these communities. Here, we link the anaerobic oxidation of methane (AOM) to the nitrogen cycle in microbial mats of the Black Sea by using stable isotope probing. We used four different (15)N-labeled sources of nitrogen: dinitrogen, nitrate, nitrite and ammonium. We estimated the nitrogen incorporation rates into the total biomass and the methyl coenzyme M reductase (MCR). Dinitrogen played an insignificant role as nitrogen source. Assimilatory and dissimilatory nitrate reduction occurred. High rates of nitrate reduction to dinitrogen were stimulated by methane and sulfate, suggesting that oxidation of reduced sulfur compounds such as sulfides was necessary for AOM with nitrate as electron acceptor. Nitrate reduction to dinitrogen occurred also in the absence of methane as electron donor but at six times slower rates. Dissimilatory nitrate reduction to ammonium was independent of AOM. Ammonium was used for biomass synthesis under all conditions. The pivotal enzyme in AOM coupled to sulfate reduction, MCR, was synthesized from nitrate and ammonium. Results show that AOM coupled to sulfate reduction along with biomass decomposition drive the nitrogen cycle in the ANME mats of the Black Sea and that MCR enzymes are involved in this process. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  1. Microbial functional diversity associated with plant litter decomposition along a climatic gradient.

    Science.gov (United States)

    Sherman, Chen; Steinberger, Yosef

    2012-08-01

    Predicted changes in climate associated with increased greenhouse gas emissions can cause increases in global mean temperature and changes in precipitation regimes. These changes may affect key soil processes, e.g., microbial CO(2) evolution and biomass, mineralization rates, primary productivity, biodiversity, and litter decomposition, which play an important role in carbon and nutrient cycling in terrestrial ecosystems. Our study examined the changes in litter microbial communities and decomposition along a climatic gradient, ranging from arid desert to humid Mediterranean regions in Israel. Wheat straw litter bags were placed in arid, semi-arid, Mediterranean, and humid Mediterranean sites. Samples were collected seasonally over a 2-year period in order to evaluate mass loss, litter moisture, C/N ratio, bacterial colony-forming units (CFUs), microbial CO(2) evolution and biomass, microbial functional diversity, and catabolic profile. Decomposition rate was the highest during the first year of the study at the Mediterranean and arid sites. Community-level physiological profile and microbial biomass were the highest in summer, while bacterial CFUs were the highest in winter. Microbial functional diversity was found to be highest at the humid Mediterranean site, whereas substrate utilization increased at the arid site. Our results support the assumption that climatic factors control litter degradation and regulate microbial activity.

  2. Diversified cropping systems support greater microbial cycling and retention of carbon and nitrogen

    Energy Technology Data Exchange (ETDEWEB)

    King, Alison E.; Hofmockel, Kirsten S.

    2017-03-01

    Diversifying biologically simple cropping systems often entails altering other management practices, such as tillage regime or nitrogen (N) source. We hypothesized that the interaction of crop rotation, N source, and tillage in diversified cropping systems would promote microbially-mediated soil C and N cycling while attenuating inorganic N pools. We studied a cropping systems trial in its 10th year in Iowa, USA, which tested a 2-yr cropping system of corn (Zea mays L.)/soybean [Glycine max (L.) Merr.] managed with conventional fertilizer N inputs and conservation tillage, a 3-yr cropping system of corn/soybean/small grain + red clover (Trifolium pratense L.), and a 4-yr cropping system of corn/soybean/small grain + alfalfa (Medicago sativa L.)/alfalfa. Three year and 4-yr cropping systems were managed with composted manure, reduced N fertilizer inputs, and periodic moldboard ploughing. We assayed soil microbial biomass carbon (MBC) and N (MBN), soil extractable NH4 and NO3, gross proteolytic activity of native soil, and potential activity of six hydrolytic enzymes eight times during the growing season. At the 0-20cm depth, native protease activity in the 4-yr cropping system was greater than in the 2-yr cropping system by a factor of 7.9, whereas dissolved inorganic N pools did not differ between cropping systems (P = 0.292). At the 0-20cm depth, MBC and MBN the 4-yr cropping system exceeded those in the 2-yr cropping system by factors of 1.51 and 1.57. Our findings suggest that diversified crop cropping systems, even when periodically moldboard ploughed, support higher levels of microbial biomass, greater production of bioavailable N from SOM, and a deeper microbially active layer than less diverse cropping systems.

  3. Recovery of microbial community structure and functioning after wildfire in semi-arid environments: optimising methods for monitoring and assessment

    Science.gov (United States)

    Muñoz-Rojas, Miriam; Martini, Dylan; Erickson, Todd; Merritt, David; Dixon, Kingsley

    2015-04-01

    Introduction In semi-arid areas such as northern Western Australia, wildfires are a natural part of the environment and many ecosystems in these landscapes have evolved and developed a strong relationship with fire. Soil microbial communities play a crucial role in ecosystem processes by regulating the cycling of nutrients via decomposition, mineralization, and immobilization processes. Thus, the structure (e.g. soil microbial biomass) and functioning (e.g. soil microbial activity) of microbial communities, as well as their changes after ecosystem disturbance, can be useful indicators of soil quality and health recovery. In this research, we assess the impacts of fire on soil microbial communities and their recovery in a biodiverse semi-arid environment of Western Australia (Pilbara region). New methods for determining soil microbial respiration as an indicator of microbial activity and soil health are also tested. Methodology Soil samples were collected from 10 similar ecosystems in the Pilbara with analogous native vegetation, but differing levels of post-fire disturbance (i.e. 3 months, 1 year, 5, 7 and 14 years after wildfire). Soil microbial activity was measured with the Solvita test which determines soil microbial respiration rate based on the measurement of the CO2 burst of a dry soil after it is moistened. Soils were dried and re-wetted and a CO2 probe was inserted before incubation at constant conditions of 25°C during 24 h. Measurements were taken with a digital mini spectrometer. Microbial (bacteria and fungi) biomass and community composition were measured by phospholipid fatty acid analysis (PLFA). Results Immediately after the fire (i.e. 3 months), soil microbial activity and microbial biomass are similar to 14 years 'undisturbed' levels (53.18±3.68 ppm CO2-CO and 14.07±0.65 mg kg-1, respectively). However, after the first year post-fire, with larger plant productivity, microbial biomass and microbial activity increase rapidly, peaking after 5

  4. Magnetic cross-linked enzyme aggregates (CLEAs): a novel concept towards carrier free immobilization of lignocellulolytic enzymes.

    Science.gov (United States)

    Bhattacharya, Abhishek; Pletschke, Brett I

    2014-01-01

    The enzymatic conversion of lignocellulosic biomass into biofuels has been identified as an excellent strategy to generate clean energy. However, the current process is cost-intensive as an effective immobilization approach to reuse the enzyme(s) has been a major challenge. The present study introduces the concept and application of novel magnetic cross-linked enzyme aggregates (mag-CLEAs). Both mag-CLEAs and calcium-mag-CLEAs (Ca-mag-CLEAs) exhibited a 1.35 fold higher xylanase activity compared to the free enzyme and retained more than 80.0% and 90.0% activity, respectively, after 136h of incubation at 50°C, compared to 50% activity retained by CLEAs. A 7.4 and 9.0 fold higher sugar release from lime-pretreated and NH4OH pre-treated sugar bagasse, respectively, was achieved with Ca-mag-CLEAs compared to the free enzymes. The present study promotes the successful application of mag-CLEAs and Ca-mag-CLEAs as carrier free immobilized enzymes for the effective hydrolysis of lignocellulolytic biomass and associated biofuel feedstocks. Copyright © 2014 Elsevier Inc. All rights reserved.

  5. Microfluidic glycosyl hydrolase screening for biomass-to-biofuel conversion.

    Science.gov (United States)

    Bharadwaj, Rajiv; Chen, Zhiwei; Datta, Supratim; Holmes, Bradley M; Sapra, Rajat; Simmons, Blake A; Adams, Paul D; Singh, Anup K

    2010-11-15

    The hydrolysis of biomass to fermentable sugars using glycosyl hydrolases such as cellulases and hemicellulases is a limiting and costly step in the conversion of biomass to biofuels. Enhancement in hydrolysis efficiency is necessary and requires improvement in both enzymes and processing strategies. Advances in both areas in turn strongly depend on the progress in developing high-throughput assays to rapidly and quantitatively screen a large number of enzymes and processing conditions. For example, the characterization of various cellodextrins and xylooligomers produced during the time course of saccharification is important in the design of suitable reactors, enzyme cocktail compositions, and biomass pretreatment schemes. We have developed a microfluidic-chip-based assay for rapid and precise characterization of glycans and xylans resulting from biomass hydrolysis. The technique enables multiplexed separation of soluble cellodextrins and xylose oligomers in around 1 min (10-fold faster than HPLC). The microfluidic device was used to elucidate the mode of action of Tm_Cel5A, a novel cellulase from hyperthermophile Thermotoga maritima . The results demonstrate that the cellulase is active at 80 °C and effectively hydrolyzes cellodextrins and ionic-liquid-pretreated switchgrass and Avicel to glucose, cellobiose, and cellotriose. The proposed microscale approach is ideal for quantitative large-scale screening of enzyme libraries for biomass hydrolysis, for development of energy feedstocks, and for polysaccharide sequencing.

  6. [Effects of simulated nitrogen deposition on soil microbial biomass carbon and nitrogen in natural evergreen broad-leaved forest in the Rainy Area of West China].

    Science.gov (United States)

    Zhou, Shi Xing; Zou, Cheng; Xiao, Yong Xiang; Xiang, Yuan Bin; Han, Bo Han; Tang, Jian Dong; Luo, Chao; Huang, Cong de

    2017-01-01

    To understand the effects of increasing nitrogen deposition on soil microbial biomass carbon (MBC) and nitrogen(MBN), an in situ experiment was conducted in a natural evergreen broad-leaved forest in Ya'an City, Sichuan Province. Four levels of nitrogen deposition were set: i.e., control (CK, 0 g N·m -2 ·a -1 ), low nitrogen (L, 5 g N·m -2 ·a -1 ), medium nitrogen (M, 15 g N·m -2 ·a -1 ), and high nitrogen (H, 30 g N·m -2 ·a -1 ). The results indicated that nitrogen deposition significantly decreased MBC and MBN in the 0-10 cm soil layer, and as N de-position increased, the inhibition effect was enhanced. L and M treatments had no significant effect on MBC and MBN in the 10-20 cm soil layer, while H treatment significantly reduced. The influence of N deposition on MBC and MBN was weakened with the increase of soil depth. MBC and MBN had obvious seasonal dynamic, which were highest in autumn and lowest in summer both in the 0-10 and 10-20 cm soil layers. The fluctuation ranges of soil microbial biomass C/N were respectively 10.58-11.19 and 9.62-12.20 in the 0-10 cm and 10-20 cm soil layers, which indicated that the fungi hold advantage in the soil microbial community in this natural evergreen broad-leaved forest.

  7. Quantitative iTRAQ secretome analysis of Aspergillus niger reveals novel hydrolytic enzymes.

    Science.gov (United States)

    Adav, Sunil S; Li, An A; Manavalan, Arulmani; Punt, Peter; Sze, Siu Kwan

    2010-08-06

    The natural lifestyle of Aspergillus niger made them more effective secretors of hydrolytic proteins and becomes critical when this species were exploited as hosts for the commercial secretion of heterologous proteins. The protein secretion profile of A. niger and its mutant at different pH was explored using iTRAQ-based quantitative proteomics approach coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS). This study characterized 102 highly confident unique proteins in the secretome with zero false discovery rate based on decoy strategy. The iTRAQ technique identified and relatively quantified many hydrolyzing enzymes such as cellulases, hemicellulases, glycoside hydrolases, proteases, peroxidases, and protein translocating transporter proteins during fermentation. The enzymes have potential application in lignocellulosic biomass hydrolysis for biofuel production, for example, the cellulolytic and hemicellulolytic enzymes glucan 1,4-alpha-glucosidase, alpha-glucosidase C, endoglucanase, alpha l-arabinofuranosidase, beta-mannosidase, glycosyl hydrolase; proteases such as tripeptidyl-peptidase, aspergillopepsin, and other enzymes including cytochrome c oxidase, cytochrome c oxidase, glucose oxidase were highly expressed in A. niger and its mutant secretion. In addition, specific enzyme production can be stimulated by controlling pH of the culture medium. Our results showed comprehensive unique secretory protein profile of A. niger, its regulation at different pH, and the potential application of iTRAQ-based quantitative proteomics for the microbial secretome analysis.

  8. Biomass degrading enzymes from Penicillium – cloning and characterization

    DEFF Research Database (Denmark)

    Krogh, Kristian Bertel Rømer

    2008-01-01

    . Størstedelen af den forskning, der er foregået indenfor cellulosenedbrydende enzymer er med enzymer produceret af svampen Trichoderma reesei. Under mit Ph.D.studium har jeg undersøgt biomassenedbrydende enzymer fra forskellige Penicillium arter. Hovedvægten af forskningen har været indenfor...... cellulosenedbrydende enzymer.Penicillium arter er blandt de hyppigst forekommende mikroorganismer i skovjord, hvori der netop nedbrydes store mængder plantemateriale. Ved en sammenligning af produktionen af biomassenedbrydende enzymer fra forskellige Penicillium arter blev der fundet flere interessante enzymsystemer...... reaktionstid ved den enzymatisk hydrolyse hvor de enkelte sukkermolekyler bliver frigivet, hvorfor enzymstabilitet er særdeles væsentlig, når et rentabelt cellulosenedbrydende enzymsystem skal sammensættes. De nødvendige enzymer for en fuldstændig hydrolyse af cellulose blev oprenset, klonet, produceret...

  9. Urease activity and its relation to soil organic matter, microbial biomass nitrogen and urea-nitrogen assimilation by maize in a Brazilian oxisol under no-tillage and tillage systems

    NARCIS (Netherlands)

    Roscoe, R.; Vasconcellos, C.A.; Furtini Neto, A.E.; Guedes, G.A.A.; Fernandes, L.A.

    2000-01-01

    We studied the relationship between urease activity (UA) and soil organic matter (SOM), microbial biomass N (Nbiom) content, and urea-N fertilizer assimilation by maize in a Dark Red Latosol (Typic Haplustox) cultivated for 9 years under no-tillage (NT), tillage with a disc plough (DP), and tillage

  10. Improved biomass utilization through the use of nuclear techniques

    International Nuclear Information System (INIS)

    1988-10-01

    Biomass is a major by-product resource of agriculture and food manufacturing, but it is under-utilized as a source of food, fibre, and chemicals. Nuclear techniques provide unique tools for studies of the capabilities of micro-organisms in methane digestor operation and in the transformation of lignocellulosic materials to useful products. Nuclear techniques have also been effectively employed as mutagenic agents in the preparation of more efficient microbial strains for the conversion of biomass. This report reviews the variety and diversity of such applications with focus on the development of microbial processes to utilize agricultural wastes and by-products. The value of nuclear techniques is manifestly demonstrated in the production of efficient microbial mutant strains, in the tracing of metabolic pathways, in the monitoring of lignin degradation and also of fermenter operation. Refs, figs and tabs

  11. [Sanitary-hygienic assessment of microbial biofertilizer].

    Science.gov (United States)

    Arkhipchenko, N A; Akhtemava, G A; Lebedeva, T V; Voronina, A A; Makhan'kova, T I; Pavlova, M M; Shteĭntsaĭg, T A

    1991-10-01

    Biological treatment of sewage from pig-breeding complexes allowed to produce microbial biomass and primary sediments. The mixture of these components (1:1) after rendering harmless and drying out become the high effective biofertilizer. The results of chronic experiment on sanitary status of soil (microbial and helminthological indexes) under this biofertilizer usage are discussed, and the harmlessness of it is demonstrated.

  12. High activity and low temperature optima of extracellular enzymes in Arctic sediments: implications for carbon cycling by heterotrophic microbial communities

    DEFF Research Database (Denmark)

    Arnosti, C.; Jørgensen, BB

    2003-01-01

    (chondroitin sulfate, fucoidan, xylan and pullulan) to determine the temperature-activity responses of hydrolysis of a related class of compounds. All 4 enzyme activities showed similarly low temperature optima in the range of 15 to 18degreesC. These temperature optima are considerably lower than most previous......The rate of the initial step in microbial remineralization of organic carbon, extracellular enzymatic hydrolysis, was investigated as a function of temperature in permanently cold sediments from 2 fjords on the west coast of Svalbard (Arctic Ocean). We used 4 structurally distinct polysaccharides...... reports of temperature optima for enzyme activities in marine sediments. At 0degreesC, close to the in situ temperature, these enzyme activities achieved 13 to 38% of their rates at optimum temperatures. In one experiment, sulfate reduction rates were measured in parallel with extracellular enzymatic...

  13. [The rise of enzyme engineering in China].

    Science.gov (United States)

    Li, Gaoxiang

    2015-06-01

    Enzyme engineering is an important part of the modern biotechnology. Industrial biocatalysis is considered the third wave of biotechnology following pharmaceutical and agricultural waves. In 25 years, China has made a mighty advances in enzyme engineering research. This review focuses on enzyme genomics, enzyme proteomics, biosynthesis, microbial conversion and biosensors in the Chinese enzyme engineering symposiums and advances in enzyme preparation industry in China.

  14. Biotechnological Processes in Microbial Amylase Production

    Directory of Open Access Journals (Sweden)

    Subash C. B. Gopinath

    2017-01-01

    Full Text Available Amylase is an important and indispensable enzyme that plays a pivotal role in the field of biotechnology. It is produced mainly from microbial sources and is used in many industries. Industrial sectors with top-down and bottom-up approaches are currently focusing on improving microbial amylase production levels by implementing bioengineering technologies. The further support of energy consumption studies, such as those on thermodynamics, pinch technology, and environment-friendly technologies, has hastened the large-scale production of the enzyme. Herein, the importance of microbial (bacteria and fungi amylase is discussed along with its production methods from the laboratory to industrial scales.

  15. Polyphenols as enzyme inhibitors in different degraded peat soils: Implication for microbial metabolism in rewetted peatlands

    Science.gov (United States)

    Zak, Dominik; Roth, Cyril; Gelbrecht, Jörg; Fenner, Nathalie; Reuter, Hendrik

    2015-04-01

    Recently, more than 30,000 ha of drained minerotrophic peatlands (= fens) in NE Germany were rewetted to restore their ecological functions. Due to an extended drainage history, a re-establishment of their original state is not expected in the short-term. Elevated concentrations of dissolved organic carbon, ammonium and phosphate have been measured in the soil porewater of the upper degraded peat layers of rewetted fens at levels of one to three orders higher than the values in pristine systems; an indicator of increased microbial activity in the upper degraded soil layers. On the other hand there is evidence that the substrate availability within the degraded peat layer is lowered since the organic matter has formerly been subject to intense decomposition over the decades of drainage and intense agricultural use of the areas. Previously however, it was suggested that inhibition of hydrolytic enzymes by polyphenolic substances is suspended during aeration of peat soils mainly due to the decomposition of the inhibiting polyphenols by oxidising enzymes such as phenol oxidase. Accordingly we hypothesised a lack of enzyme inhibiting polyphenols in degraded peat soils of rewetted fens compared to less decomposed peat of more natural fens. We collected both peat samples at the soil surface (0-20 cm) and fresh roots of dominating vascular plants and mosses (as peat parent material) from five formerly drained rewetted sites and five more natural sites of NE Germany and NW Poland. Less decomposed peat and living roots were used to obtain an internal standard for polyphenol analysis and to run enzyme inhibition tests. For all samples we determined the total phenolic contents and in addition we distinguished between the contents of hydrolysable and condensed tannic substances. From a methodical perspective the advantage of internal standards compared to the commercially available standards cyanidin chloride and tannic acid became apparent. Quantification with cyanidin or

  16. Microbially produced phytotoxins and plant disease management ...

    African Journals Online (AJOL)

    Nowadays, these evaluation techniques are becoming an important complement to classical breeding methods. The knowledge of the inactivation of microbial toxins has led to the use of microbial enzymes to inactivate phytotoxins thereby reducing incidence and severity of disease induced by microbial toxins. Considering ...

  17. Exploring Biogeochemistry and Microbial Diversity of Extant Microbialites in Mexico and Cuba

    Science.gov (United States)

    Valdespino-Castillo, Patricia M.; Hu, Ping; Merino-Ibarra, Martín; López-Gómez, Luz M.; Cerqueda-García, Daniel; González-De Zayas, Roberto; Pi-Puig, Teresa; Lestayo, Julio A.; Holman, Hoi-Ying; Falcón, Luisa I.

    2018-01-01

    Microbialites are modern analogs of ancient microbial consortia that date as far back as the Archaean Eon. Microbialites have contributed to the geochemical history of our planet through their diverse metabolic capacities that mediate mineral precipitation. These mineral-forming microbial assemblages accumulate major ions, trace elements and biomass from their ambient aquatic environments; their role in the resulting chemical structure of these lithifications needs clarification. We studied the biogeochemistry and microbial structure of microbialites collected from diverse locations in Mexico and in a previously undescribed microbialite in Cuba. We examined their structure, chemistry and mineralogy at different scales using an array of nested methods including 16S rRNA gene high-throughput sequencing, elemental analysis, X-Ray fluorescence (XRF), X-Ray diffraction (XRD), Scanning Electron Microscopy-Energy Dispersive Spectroscopy (SEM-EDS), Fourier Transformed Infrared (FTIR) spectroscopy and Synchrotron Radiation-based Fourier Transformed Infrared (SR-FTIR) spectromicroscopy. The resulting data revealed high biological and chemical diversity among microbialites and specific microbe to chemical correlations. Regardless of the sampling site, Proteobacteria had the most significant correlations with biogeochemical parameters such as organic carbon (Corg), nitrogen and Corg:Ca ratio. Biogeochemically relevant bacterial groups (dominant phototrophs and heterotrophs) showed significant correlations with major ion composition, mineral type and transition element content, such as cadmium, cobalt, chromium, copper and nickel. Microbial-chemical relationships were discussed in reference to microbialite formation, microbial metabolic capacities and the role of transition elements as enzyme cofactors. This paper provides an analytical baseline to drive our understanding of the links between microbial diversity with the chemistry of their lithified precipitations. PMID

  18. Are variations in heterotrophic soil respiration related to changes in substrate availability and microbial biomass carbon in the subtropical forests?

    Science.gov (United States)

    Wei, Hui; Chen, Xiaomei; Xiao, Guoliang; Guenet, Bertrand; Vicca, Sara; Shen, Weijun

    2015-01-01

    Soil temperature and moisture are widely-recognized controlling factors on heterotrophic soil respiration (Rh), although they often explain only a portion of Rh variability. How other soil physicochemical and microbial properties may contribute to Rh variability has been less studied. We conducted field measurements on Rh half-monthly and associated soil properties monthly for two years in four subtropical forests of southern China to assess influences of carbon availability and microbial properties on Rh. Rh in coniferous forest was significantly lower than that in the other three broadleaf species-dominated forests and exhibited obvious seasonal variations in the four forests (P < 0.05). Temperature was the primary factor influencing the seasonal variability of Rh while moisture was not in these humid subtropical forests. The quantity and decomposability of dissolved organic carbon (DOC) were significantly important to Rh variations, but the effect of DOC content on Rh was confounded with temperature, as revealed by partial mantel test. Microbial biomass carbon (MBC) was significantly related to Rh variations across forests during the warm season (P = 0.043). Our results suggest that DOC and MBC may be important when predicting Rh under some conditions, and highlight the complexity by mutual effects of them with environmental factors on Rh variations. PMID:26670822

  19. Black Nitrogen as a source for the built-up of microbial biomass in soils

    Science.gov (United States)

    López-Martín, María; Milter, Anja; Knicker, Heike

    2016-04-01

    In areas with frequent wildfires, soil organic nitrogen (SON) is sequestered in pyrogenic organic matter (PyOM) due to heat-induced transformation of proteinaceous compounds into N-heterocycles, i.e. pyrrole, imidazole and indole compounds. These newly formed structures, known as Black Nitrogen (BN), have been assumed to be hardly degradable by microorganisms, thus being efficiently sequestered from the N cycle. On the other hand, a previous study showed that nitrogen of BN can be used by plants for the built-up of their biomass (de la Rosa and Knicker 2011). Thus, BN may play an important role as an N source during the recovery of the forest after a fire event. In order to obtain a more profound understanding of the role of BN within the N cycle in soils, we studied the bioavailability and incorporation of N derived from PyOM into microbial amino acids. For that, pots with soil from a burnt and an unburnt Cambisol located under a Mediterranean forest were covered with different amendments. The toppings were mixtures of unlabeled KNO3 with 15N labeled grass or 15N-labeled PyOM from burned grass and K15NO3 mixed with unlabeled grass material or PyOM. The pots were kept in the greenhouse under controlled conditions for 16 months and were sampled after 0.5, 1, 5, 8 and 16 months. From all samples the amino acids were extracted after hydrolysis (6 M HCl, 22 h, 110 °C) and quantified via gas chromatography mass spectrometry (GC/MS). The fate of 15N was followed by isotopic ratio mass spectrometry (IRMS). The results show that the contribution of extractable amino acids to total soil organic matter was always higher in the unburnt than in the burnt soil. However, with ongoing incubation their amount decreased. Already after 0.5 months, some PyOM-derived 15N was incorporated into the extractable amino acids and the amount increased with experiment time. Since this can only occur after prior microbial degradation of PyOM our results clearly support a lower biochemical

  20. Dissolved nitrogen transformations and microbial community structure in the organic layer of forest soils in Olkiluoto in 2006

    International Nuclear Information System (INIS)

    Potila, H.; Sarjala, T.; Aro, L.

    2007-02-01

    Carbon (C) and nitrogen (N) cycles in the ecosystem are strongly coupled. Biomass, structure and activity of the bacterial and fungal community are the key factors influencing C and N cycles. Changes in the function of soil microbial community can be a signal of plant responses to environmental changes. Dissolved N compounds, microbial biomass, microbial activity, fungal community structure and functional diversity of microbial communities were measured in September 2006 from five monitoring plots on Olkiluoto to assess information about soil microbial community structure and activity. High within and between variation in the studied plots were detected. However, in this study the values and their variation in the level of N mineralisation, dissolved N compounds, fungal biomass and microbial community structure in the studied plots were within a normal range in comparison with other published data of similar forest types in Finland. (orig.)

  1. Enzyme Activities in Waste Water and Activated Sludge

    DEFF Research Database (Denmark)

    Nybroe, Ole; Jørgensen, Per Elberg; Henze, Mogens

    1992-01-01

    The purpose of the present study was to evaluate the potential of selected enzyme activity assays to determine microbial abundance and heterotrophic activity in waste water and activated sludge. In waste water, esterase and dehydrogenase activities were found to correlate with microbial abundance...... measured as colony forming units of heterotrophic bacteria. A panel of four enzyme activity assays, α-glucosidase, alanine-aminopeptidase, esterase and dehydrogenase were used to characterize activated sludge and anaerobic hydrolysis sludge from a pilot scale plant. The enzymatic activity profiles were...... distinctly different, suggesting that microbial populations were different, or had different physiological properties, in the two types of sludge. Enzyme activity profiles in activated sludge from four full-scale plants seemed to be highly influenced by the composition of the inlet. Addition of hydrolysed...

  2. Geographic variability in amoeboid protists and other microbial groups in the water column of the lower Hudson River Estuary (New York, USA)

    Science.gov (United States)

    Juhl, Andrew R.; Anderson, O. Roger

    2014-12-01

    In comparison to other groups of planktonic microorganisms, relatively little is known about the role of amoeboid protists (amebas) in planktonic ecosystems. This study describes the first geographic survey of the abundance and biomass of amebas in an estuarine water column. Samples collected in the lower Hudson River Estuary were used to investigate relationships between ameba abundance and biomass and hydrographic variables (temperature, salinity, and turbidity), water depth (surface and near bottom), distance from mid-channel to shore, phytoplankton biomass (chlorophyll fluorescence) and the occurrence of other heterotrophic microbial groups (heterotrophic bacteria, nanoflagellates, and ciliates) in the plankton. Although salinity increased significantly towards the mouth of the estuary, there were no significant differences in the abundance or biomass of any microbial group in surface samples collected at three stations separated by 44 km along the estuary's mid-channel. Peak biomass values for all microbial groups were found at the station closest to shore, however, cross-channel trends in microbial abundance and biomass were not statistically significant. Although ameba abundance and biomass in most samples were low compared to other microbial groups, clear patterns in ameba distribution were nevertheless found. Unlike other microbial groups examined, ameba numbers and biomass greatly increased in near bottom water compared to surface samples. Ameba abundance and biomass (in surface samples) were also strongly related to increasing turbidity. The different relationships of ameba abundance and biomass with turbidity suggest a rising contribution of large amebas in microbial communities of the Hudson estuary when turbidity increases. These results, emphasizing the importance of particle concentration as attachment and feeding surfaces for amebas, will help identify the environmental conditions when amebas are most likely to contribute significantly to estuarine

  3. Production of Microbial Protease from Selected Soil Fungal Isolates ...

    African Journals Online (AJOL)

    Production of Microbial Protease from Selected Soil Fungal Isolates. ... Nigerian Journal of Biotechnology ... and 500C. The optimal pH on the enzyme production was observed to be between pH 3.5 and 5.5 for the organisms. Keywords: Soil microorganism, fungal isolate, incubation period, microbial enzyme. Nig J. Biotech.

  4. Defining Disturbance for Microbial Ecology.

    Science.gov (United States)

    Plante, Craig J

    2017-08-01

    Disturbance can profoundly modify the structure of natural communities. However, microbial ecologists' concept of "disturbance" has often deviated from conventional practice. Definitions (or implicit usage) have frequently included climate change and other forms of chronic environmental stress, which contradict the macrobiologist's notion of disturbance as a discrete event that removes biomass. Physical constraints and disparate biological characteristics were compared to ask whether disturbances fundamentally differ in microbial and macroorganismal communities. A definition of "disturbance" for microbial ecologists is proposed that distinguishes from "stress" and other competing terms, and that is in accord with definitions accepted by plant and animal ecologists.

  5. Cellulase digestibility of pretreated biomass is limited by cellulose accessibility.

    Science.gov (United States)

    Jeoh, Tina; Ishizawa, Claudia I; Davis, Mark F; Himmel, Michael E; Adney, William S; Johnson, David K

    2007-09-01

    Attempts to correlate the physical and chemical properties of biomass to its susceptibility to enzyme digestion are often inconclusive or contradictory depending on variables such as the type of substrate, the pretreatment conditions and measurement techniques. In this study, we present a direct method for measuring the key factors governing cellulose digestibility in a biomass sample by directly probing cellulase binding and activity using a purified cellobiohydrolase (Cel7A) from Trichoderma reesei. Fluorescence-labeled T. reesei Cel7A was used to assay pretreated corn stover samples and pure cellulosic substrates to identify barriers to accessibility by this important component of cellulase preparations. The results showed cellulose conversion improved when T. reesei Cel7A bound in higher concentrations, indicating that the enzyme had greater access to the substrate. Factors such as the pretreatment severity, drying after pretreatment, and cellulose crystallinity were found to directly impact enzyme accessibility. This study provides direct evidence to support the notion that the best pretreatment schemes for rendering biomass more digestible to cellobiohydrolase enzymes are those that improve access to the cellulose in biomass cell walls, as well as those able to reduce the crystallinity of cell wall cellulose.

  6. Increasing atmospheric deposition nitrogen and ammonium reduced microbial activity and changed the bacterial community composition of red paddy soil.

    Science.gov (United States)

    Zhou, Fengwu; Cui, Jian; Zhou, Jing; Yang, John; Li, Yong; Leng, Qiangmei; Wang, Yangqing; He, Dongyi; Song, Liyan; Gao, Min; Zeng, Jun; Chan, Andy

    2018-03-27

    Atmospheric deposition nitrogen (ADN) increases the N content in soil and subsequently impacts microbial activity of soil. However, the effects of ADN on paddy soil microbial activity have not been well characterized. In this study, we studied how red paddy soil microbial activity responses to different contents of ADN through a 10-months ADN simulation on well managed pot experiments. Results showed that all tested contents of ADN fluxes (27, 55, and 82kgNha -1 when its ratio of NH 4 + /NO 3 - -N (R N ) was 2:1) enhanced the soil enzyme activity and microbial biomass carbon and nitrogen and 27kgNha -1 ADN had maximum effects while comparing with the fertilizer treatment. Generally, increasing of both ADN flux and R N (1:2, 1:1 and 2:1 with the ADN flux of 55kgNha -1 ) had similar reduced effects on microbial activity. Furthermore, both ADN flux and R N significantly reduced soil bacterial alpha diversity (pADN flux and R N were the main drivers in shaping paddy soil bacteria community. Overall, the results have indicated that increasing ADN flux and ammonium reduced soil microbial activity and changed the soil bacterial community. The finding highlights how paddy soil microbial community response to ADN and provides information for N management in paddy soil. Copyright © 2018 Elsevier B.V. All rights reserved.

  7. Soil bacterial and fungal diversity differently correlated with soil biochemistry in alpine grassland ecosystems in response to environmental changes

    Science.gov (United States)

    Zhang, Yong; Dong, Shikui; Gao, Qingzhu; Liu, Shiliang; Ganjurjav, Hasbagan; Wang, Xuexia; Su, Xukun; Wu, Xiaoyu

    2017-03-01

    To understand effects of soil microbes on soil biochemistry in alpine grassland ecosystems under environmental changes, we explored relationships between soil microbial diversity and soil total nitrogen, organic carbon, available nitrogen and phosphorus, soil microbial biomass and soil enzyme activities in alpine meadow, alpine steppe and cultivated grassland on the Qinghai-Tibetan plateau under three-year warming, enhanced precipitation and yak overgrazing. Soil total nitrogen, organic carbon and NH4-N were little affected by overgrazing, warming or enhanced precipitation in three types of alpine grasslands. Soil microbial biomass carbon and phosphorus along with the sucrase and phosphatase activities were generally stable under different treatments. Soil NO3-N, available phosphorus, urease activity and microbial biomass nitrogen were increased by overgrazing in the cultivated grassland. Soil bacterial diversity was positively correlated with, while soil fungal diversity negatively with soil microbial biomass and enzyme activities. Soil bacterial diversity was negatively correlated with, while soil fungal diversity positively with soil available nutrients. Our findings indicated soil bacteria and fungi played different roles in affecting soil nutrients and microbiological activities that might provide an important implication to understand why soil biochemistry was generally stable under environmental changes in alpine grassland ecosystems.

  8. High-throughput microplate technique for enzymatic hydrolysis of lignocellulosic biomass.

    Science.gov (United States)

    Chundawat, Shishir P S; Balan, Venkatesh; Dale, Bruce E

    2008-04-15

    Several factors will influence the viability of a biochemical platform for manufacturing lignocellulosic based fuels and chemicals, for example, genetically engineering energy crops, reducing pre-treatment severity, and minimizing enzyme loading. Past research on biomass conversion has focused largely on acid based pre-treatment technologies that fractionate lignin and hemicellulose from cellulose. However, for alkaline based (e.g., AFEX) and other lower severity pre-treatments it becomes critical to co-hydrolyze cellulose and hemicellulose using an optimized enzyme cocktail. Lignocellulosics are appropriate substrates to assess hydrolytic activity of enzyme mixtures compared to conventional unrealistic substrates (e.g., filter paper, chromogenic, and fluorigenic compounds) for studying synergistic hydrolysis. However, there are few, if any, high-throughput lignocellulosic digestibility analytical platforms for optimizing biomass conversion. The 96-well Biomass Conversion Research Lab (BCRL) microplate method is a high-throughput assay to study digestibility of lignocellulosic biomass as a function of biomass composition, pre-treatment severity, and enzyme composition. The most suitable method for delivering milled biomass to the microplate was through multi-pipetting slurry suspensions. A rapid bio-enzymatic, spectrophotometric assay was used to determine fermentable sugars. The entire procedure was automated using a robotic pipetting workstation. Several parameters that affect hydrolysis in the microplate were studied and optimized (i.e., particle size reduction, slurry solids concentration, glucan loading, mass transfer issues, and time period for hydrolysis). The microplate method was optimized for crystalline cellulose (Avicel) and ammonia fiber expansion (AFEX) pre-treated corn stover. Copyright 2008 Wiley Periodicals, Inc.

  9. Microbial photosynthesis in the harnessing of solar energy

    Energy Technology Data Exchange (ETDEWEB)

    Pirt, S J

    1982-01-01

    The shortage of fossil fuels restricts the world supply of reduced carbon compounds and energy sources. Biotechnology offers the most feasible route to renewing the supplies of reduced carbon compounds. This involves recycling of CO/sub 2/ through photosynthesis. Conventional agriculture has little or no potential for supplying biomass and its derivatives on sufficient scale to offer an alternative to the fossil fuels. The agricultural wastes, on the whole, are intractable to conversion into useful carbon and energy sources and in any case are not available in amounts to provide a significant alternative to the fossil fuels. In contrast, microbial photosynthesis, optimised in photobioreactors, has vast potential to provide organic matter on a scale to match the consumption of fossil fuels. The quantative study of microbial photosynthesis as a biotechnological route to biomass has been neglected. As a result there is a chaos of conflicting data on fundamental parameters, for example, the photosynthetic efficiency of biomass production. New photosynthetic biotechnology with fully controlled continuous-culture systems is providing unequivocal values for the parameters. For the scale-up of microbial photosynthesis a tubular-loop reactor is proposed. (Refs. 14).

  10. Consolidated Bio-Processing of Cellulosic Biomass for Efficient Biofuel Production Using Yeast Consortium

    Science.gov (United States)

    Goyal, Garima

    Fossil fuels have been the major source for liquid transportation fuels for ages. However, decline in oil reserves and environmental concerns have raised a lot of interest in alternative and renewable energy sources. One promising alternative is the conversion of plant biomass into ethanol. The primary biomass feed stocks currently being used for the ethanol industry have been food based biomass (corn and sugar cane). However, interest has recently shifted to replace these traditional feed-stocks with more abundant, non-food based cellulosic biomass such as agriculture wastes (corn stover) or crops (switch grass). The use of cellulosic biomass as feed stock for the production of ethanol via bio-chemical routes presents many technical challenges not faced with the use of corn or sugar-cane as feed-stock. Recently, a new process called consolidated Bio-processing (CBP) has been proposed. This process combines simultaneous saccharification of lignocellulose with fermentation of the resulting sugars into a single process step mediated by a single microorganism or microbial consortium. Although there is no natural microorganism that possesses all properties of lignocellulose utilization and ethanol production desired for CBP, some bacteria and fungi exhibit some of the essential traits. The yeast Saccharomyces cerevisiae is the most attractive host organism for the usage of this strategy due to its high ethanol productivity at close to theoretical yields (0.51g ethanol/g glucose consumed), high osmo- and ethanol- tolerance, natural robustness in industrial processes, and ease of genetic manipulation. Introduction of the cellulosome, found naturally in microorganisms, has shown new directions to deal with recalcitrant biomass. In this case enzymes work in synergy in order to hydrolyze biomass more effectively than in case of free enzymes. A microbial consortium has been successfully developed, which ensures the functional assembly of minicellulosome on the yeast surface

  11. Effect of Arbuscular Mycorrhizal Fungi on Plant Biomass and the Rhizosphere Microbial Community Structure of Mesquite Grown in Acidic Lead/Zinc Mine Tailings

    Science.gov (United States)

    Solís-Domínguez, Fernando A.; Valentín-Vargas, Alexis; Chorover, Jon; Maier, Raina M.

    2011-01-01

    Mine tailings in arid and semi-arid environments are barren of vegetation and subject to eolian dispersion and water erosion. Revegetation is a cost-effective strategy to reduce erosion processes and has wide public acceptance. A major cost of revegetation is the addition of amendments, such as compost, to allow plant establishment. In this paper we explore whether arbuscular mycorrhizal fungi (AMF) can help support plant growth in tailings at a reduced compost concentration. A greenhouse experiment was performed to determine the effects of three AMF inocula on biomass, shoot accumulation of heavy metals, and changes in the rhizosphere microbial community structure of the native plant Prosopis juliflora (mesquite). Plants were grown in an acidic lead/zinc mine tailings amended with 10% (w/w) compost amendment, which is slightly sub-optimal for plant growth in these tailings. After two months, AMF-inoculated plants showed increased dry biomass and root length (p tailings. Mesquite shoot tissue lead and zinc concentrations did not exceed domestic animal toxicity limits regardless of whether AMF inoculation was used. The rhizosphere microbial community structure was assessed using denaturing gradient gel electrophoresis (DGGE) profiles of the small subunit RNA gene for bacteria and fungi. Canonical correspondence analysis (CCA) of DGGE profiles showed that the rhizosphere fungal community structure at the end of the experiment was significantly different from the community structure in the tailings, compost, and AMF inocula prior to planting. Further, CCA showed that AMF inoculation significantly influenced the development of both the fungal and bacterial rhizosphere community structures after two months. The changes observed in the rhizosphere microbial community structure may be either a direct effect of the AMF inocula, caused by changes in plant physiology induced by AMF, or a combination of both mechanisms. PMID:21211826

  12. Microbial amylases in the production of alcohol

    Energy Technology Data Exchange (ETDEWEB)

    Pieper, H J

    1970-01-01

    This book is based on experiments carried out in the experimental distillery of the University of Hohenheim on the use of microbial enzyme preparations for processing wheat and maize, with particular reference to comparison of green and cured malts. The subject is divided into the following chapters: introduction (pp. -14); raw materials (pp. 5-6); enzymic dextrinizing and saccharification agents (pp. 6-10); technology of alcohol production with microbial amylses (pp. 11-27); experiments into, results of and discussion on special problems of the mashing and fermentation process with reference to application of microbial amylases (pp. 28-45); Analytical methods (pp. 46-51); and Resume (pp. 5254).

  13. Effects of simulated acid rain on microbial characteristics in a lateritic red soil.

    Science.gov (United States)

    Xu, Hua-qin; Zhang, Jia-en; Ouyang, Ying; Lin, Ling; Quan, Guo-ming; Zhao, Ben-liang; Yu, Jia-yu

    2015-11-01

    A laboratory experiment was performed to examine the impact of simulated acid rain (SAR) on nutrient leaching, microbial biomass, and microbial activities in a lateritic red soil in South China. The soil column leaching experiment was conducted over a 60-day period with the following six SAR pH treatments (levels): 2.5, 3.0, 3.5, 4.0, 4.5, and 5.0 and one control treatment (pH = 7). Compared with the control treatment, the concentrations of soil organic matter, total nitrogen, total phosphorus, total potassium, soil microbial biomass carbon (MBC), soil microbial biomass nitrogen (MBN), and average well color density (AWCD) in the Ecoplates were all significantly decreased by leaching with SAR at different pH levels. The decrease in MBC and MBN indicated that acid rain reduced the soil microbial population, while the decrease in AWCD revealed that acid rain had a negative effect on soil bacterial metabolic function. Soil basal respiration increased gradually from pH 4.0 to 7.0 but decreased dramatically from pH 2.5 to 3.0. The decrease in soil nutrient was the major reason for the change of soil microbial functions. A principal component analysis showed that the major carbon sources used by the bacteria were carbohydrates and carboxylic acids.

  14. Adapting enzyme-based microbial water quality analysis to remote areas in low-income countries.

    Science.gov (United States)

    Abramson, Adam; Benami, Maya; Weisbrod, Noam

    2013-09-17

    Enzyme-substrate microbial water tests, originally developed for efficiency gains in laboratory settings, are potentially useful for on-site analysis in remote settings. This is especially relevant in developing countries where water quality is a pressing concern and qualified laboratories are rare. We investigated one such method, Colisure, first for sensitivity to incubation temperatures in order to explore alternative incubation techniques appropriate for remote areas, and then in a remote community of Zambia for detection of total coliforms and Escherichia coli in drinking-water samples. We sampled and analyzed 352 water samples from source, transport containers and point-of-use from 164 random households. Both internal validity (96-100%) and laboratory trials (zero false negatives or positives at incubation between 30 and 40 °C) established reliability under field conditions. We therefore recommend the use of this and other enzyme-based methods for remote applications. We also found that most water samples from wells accessing groundwater were free of E. coli whereas most samples from surface sources were fecally contaminated. We further found very low awareness among the population of the high levels of recontamination in household storage containers, suggesting the need for monitoring and treatment beyond the water source itself.

  15. Effects of resource chemistry on the composition and function of stream hyporheic biofilms.

    Science.gov (United States)

    Hall, E.K.; Besemer, K.; Kohl, L.; Preiler, C.; Reidel, K.; Schneider, T.; Wanek, W.; Battin, T.J.

    2012-01-01

    Fluvial ecosystems process large quantities of dissolved organic matter as it moves from the headwater streams to the sea. In particular, hyporheic sediments are centers of high biogeochemical reactivity due to their elevated residence time and high microbial biomass and activity. However, the interaction between organic matter and microbial dynamics in the hyporheic zone remains poorly understood. We evaluated how variance in resource chemistry affected the microbial community and its associated activity in experimentally grown hyporheic biofilms. To do this we fed beech leaf leachates that differed in chemical composition to a series of bioreactors filled with sediment from a sub-alpine stream. Differences in resource chemistry resulted in differences in diversity and phylogenetic origin of microbial proteins, enzyme activity, and microbial biomass stoichiometry. Specifically, increased lignin, phenolics, and manganese in a single leachate resulted in increased phenoloxidase and peroxidase activity, elevated microbial biomass carbon:nitrogen ratio, and a greater proportion of proteins of Betaproteobacteria origin. We used this model system to attempt to link microbial form (community composition and metaproteome) with function (enzyme activity) in order to better understand the mechanisms that link resource heterogeneity to ecosystem function in stream ecosystems.

  16. Effects of resource chemistry on the composition and function of hyporheic stream biofilms

    Directory of Open Access Journals (Sweden)

    Edward eHall

    2012-02-01

    Full Text Available Stream ecosystems process large quantities of dissolved organic matter as it moves from the headwaters to the sea. Interstitial sediments in the hyporheic zone are centers of high biogeochemical reactivity due to their high levels of microbial biomass and activity. However, the interaction between organic matter and microbial dynamics of these systems remains poorly understood. We evaluated how variance in resource chemistry affected the microbial community and its associated activity in experimentally grown interstitial biofilms. Specifically, we fed beech leaf leachates that differed in chemical composition to a series of bioreactors filled with sediment from a sub-alpine stream. Differences in resource chemistry resulted in differences in diversity and phylogenetic origin of microbial proteins, enzyme activity, and microbial biomass stoichiometry. Specifically, increased lignin, phenolics and manganese in a single leachate resulted in increased phenoloxidase and peroxidase activity, elevated microbial biomass carbon:nitrogen ratio, and a greater proportion of proteins of beta-proteobacter origin. We use this model system to link microbial form, (community composition and proteome, with function, (enzyme activity, in an attempt to develop a better understanding of the mechanisms that link resource heterogeneity to ecosystem function in stream ecosystems.

  17. In-depth investigation of enzymatic hydrolysis of biomass wastes based on three major components: Cellulose, hemicellulose and lignin.

    Science.gov (United States)

    Lin, Lili; Yan, Rong; Liu, Yongqiang; Jiang, Wenju

    2010-11-01

    The artificial biomass based on three biomass components (cellulose, hemicellulose and lignin) were developed on the basis of a simplex-lattice approach. Together with a natural biomass sample, they were employed in enzymatic hydrolysis researches. Different enzyme combines of two commercial enzymes (ACCELLERASE 1500 and OPTIMASH BG) showed a potential to hydrolyze hemicellulose completely. Negligible interactions among the three components were observed, and the used enzyme ACCELLERASE 1500 was proven to be weak lignin-binding. On this basis, a multiple linear-regression equation was established for predicting the reducing sugar yield based on the component proportions in a biomass. The hemicellulose and cellulose in a biomass sample were found to have different contributions in staged hydrolysis at different time periods. Furthermore, the hydrolysis of rice straw was conducted to validate the computation approach through considerations of alkaline solution pretreatment and combined enzymes function, so as to understand better the nature of biomass hydrolysis, from the aspect of three biomass components.

  18. Differential Responses of Soil Microbial Community to Four-Decade Long Grazing and Cultivation in a Semi-Arid Grassland

    Directory of Open Access Journals (Sweden)

    Yating He

    2017-01-01

    Full Text Available Grazing and cultivation are two important management practices worldwide that can cause significant soil organic carbon (SOC losses. However, it remains elusive how soil microbes have responded to soil carbon changes under these two practices. Based on a four-decade long field experiment, this study investigated the effects of grazing and cultivation on SOC stocks and microbial properties in the semi-arid grasslands of China. We hypothesize that grazing and cultivation would deplete SOC and depress microbial activities under both practices. However, our hypotheses were only partially supported. As compared with the adjacent indigenous grasslands, SOC and microbial biomass carbon (MBC were decreased by 20% or more under grazing and cultivation, which is consistent with the reduction of fungi abundance by 40% and 71%, respectively. The abundance of bacteria and actinomycetes was decreased under grazing but increased under cultivation, which likely enhanced microbial diversity in cultivation. Invertase activity decreased under the two treatments, while urease activity increased under grazing. These results suggest that nitrogen fertilizer input during cultivation may preferentially favor bacterial growth, in spite of SOC loss, due to rapid decomposition, while overgrazing may deteriorate the nitrogen supply to belowground microbes, thus stimulating the microbial production of nitrogen acquisition enzymes. This decade-long study demonstrated differential soil microbial responses under grazing and cultivation and has important applications for better management practices in the grassland ecosystem.

  19. Enzymes of industrial purpose - review of the market of enzyme preparations and prospects for its development

    Directory of Open Access Journals (Sweden)

    A. A. Tolkacheva

    2017-01-01

    Full Text Available Microbial enzyme preparations are increasingly replacing conventional chemical catalysts in a number of industrial processes. Such drugs, in addition to environmental friendliness and high activity, have a number of advantages over enzyme preparations of vegetable and animal origin, namely: the production of microbial enzymes in bioreactors is easily controlled and predictable; excreted microbiological enzymes are more stable than intracellular animals and plant enzymes; the genetic diversity of microorganisms makes it possible to produce enzyme preparations with a wide range of specificity; microbiological enzymes can be synthesized year-round, in contrast to the production of plant enzymes, which is often seasonal. The leaders of the world market of enzymes are proteases and amylases, which account for 25% and 15%, respectively. Over the past five years, the world market for carbohydrases, including mainly amylases, cellulases and xylanases, has been the fastest growing segment of the enzyme market with an aggregate annual growth rate of more than 7.0%. Another major product of the industrial enzyme market, which has a great potential for growth, is lipases. From the point of view of designation, the main part is represented by food and food enzymes. The Russian market continues to be unsaturated - the current supply is not able to meet the needs of the Russian feed and food industry in enzyme preparations. Enzyme preparations of domestic producers are in demand in forage production, while food industrial enterprises prefer imported products. The most significant enterprises in the enzymatic industry in Russia at the moment are Sibbiofarm, AgroSistema, Agroferment. In the light of the Russian policy of increasing food security, the development of the domestic enzyme industry is an extremely topical task.

  20. Visualizing Microbial Biogeochemistry: NanoSIMS and Stable Isotope Probing (Invited)

    Science.gov (United States)

    Pett-Ridge, J.; Weber, P. K.

    2009-12-01

    Linking phylogenetic information to function in microbial communities is a key challenge for microbial ecology. Isotope-labeling experiments provide a useful means to investigate the ecophysiology of microbial populations and cells in the environment and allow measurement of nutrient transfers between cell types, symbionts and consortia. The combination of Nano-Secondary Ion Mass Spectrometry (NanoSIMS) analysis, in situ labeling and high resolution microscopy allows isotopic analysis to be linked to phylogeny and morphology and holds great promise for fine-scale studies of microbial systems. In NanoSIMS analysis, samples are sputtered with an energetic primary beam (Cs+, O-) liberating secondary ions that are separated by the mass spectrometer and detected in a suite of electron multipliers. Five isotopic species may be analyzed concurrently with spatial resolution as fine as 50nm. A high sensitivity isotope ratio ‘map’ can then be generated for the analyzed area. NanoSIMS images of 13C, 15N and Mo (a nitrogenase co-factor) localization in diazotrophic cyanobacteria show how cells differentially allocate resources within filaments and allow calculation of nutrient uptake rates on a cell by cell basis. Images of AM fungal hyphae-root and cyanobacteria-rhizobia associations indicate the mobilization and sharing (stealing?) of newly fixed C and N. In a related technique, “El-FISH”, stable isotope labeled biomass is probed with oligonucleotide-elemental labels and then imaged by NanoSIMS. In microbial consortia and cyanobacterial mats, this technique helps link microbial structure and function simultaneously even in systems with unknown and uncultivated microbes. Finally, the combination of re-engineered universal 16S oligonucleotide microarrays with NanoSIMS analyses may allow microbial identity to be linked to functional roles in complex systems such as mats and cellulose degrading hindgut communities. These newly developed methods provide correlated

  1. To prevent the occurrence of black water agglomerate through delaying decomposition of cyanobacterial bloom biomass by sediment microbial fuel cell.

    Science.gov (United States)

    Zhou, Yan-Li; Jiang, He-Long; Cai, Hai-Yuan

    2015-04-28

    Settlement of cyanobacterial bloom biomass (CBB) into sediments in eutrophic lakes often induced the occurrence of black water agglomerate and then water quality deterioration. This study investigated the effect of sediment microbial fuel cell (SMFC) on CBB removal in sediments and related water pollution. Sediment bulking and subsequent black water from decomposition of settled CBB happened without SMFC, but were not observed over 100-day experiments with SMFC employment. While CBB in sediments improved power production from SMFC, the removal efficiency of organic matters in CBB-amended sediments with SMFC was significantly lower than that without SMFC. Pyrosequencing analysis showed higher abundances of the fermentative Clostridium and acetoclastic methanogen in CBB-amended bulk sediments without SMFC than with SMFC at the end of experiments. Obviously, SMFC operation changed the microbial community in CBB-amended sediments, and delayed the CBB degradation against sediment bulking. Thus, SMFC could be potentially applied as pollution prevention in CBB-settled and sensitive zones in shallow lakes. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. Biochar affects soil organic matter cycling and microbial functions but does not alter microbial community structure in a paddy soil.

    Science.gov (United States)

    Tian, Jing; Wang, Jingyuan; Dippold, Michaela; Gao, Yang; Blagodatskaya, Evgenia; Kuzyakov, Yakov

    2016-06-15

    The application of biochar (BC) in conjunction with mineral fertilizers is one of the most promising management practices recommended to improve soil quality. However, the interactive mechanisms of BC and mineral fertilizer addition affecting microbial communities and functions associated with soil organic matter (SOM) cycling are poorly understood. We investigated the SOM in physical and chemical fractions, microbial community structure (using phospholipid fatty acid analysis, PLFA) and functions (by analyzing enzymes involved in C and N cycling and Biolog) in a 6-year field experiment with BC and NPK amendment. BC application increased total soil C and particulate organic C for 47.4-50.4% and 63.7-74.6%, respectively. The effects of BC on the microbial community and C-cycling enzymes were dependent on fertilization. Addition of BC alone did not change the microbial community compared with the control, but altered the microbial community structure in conjunction with NPK fertilization. SOM fractions accounted for 55% of the variance in the PLFA-related microbial community structure. The particulate organic N explained the largest variation in the microbial community structure. Microbial metabolic activity strongly increased after BC addition, particularly the utilization of amino acids and amines due to an increase in the activity of proteolytic (l-leucine aminopeptidase) enzymes. These results indicate that microorganisms start to mine N from the SOM to compensate for high C:N ratios after BC application, which consequently accelerate cycling of stable N. Concluding, BC in combination with NPK fertilizer application strongly affected microbial community composition and functions, which consequently influenced SOM cycling. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Toxic effect of two kinds of mineral collectors on soil microbial richness and activity: analysis by microcalorimetry, microbial count, and enzyme activity assay.

    Science.gov (United States)

    Bararunyeretse, Prudence; Yao, Jun; Dai, Yunrong; Bigawa, Samuel; Guo, Zunwei; Zhu, Mijia

    2017-01-01

    Flotation reagents are hugely and increasingly used in mining and other industrial and economic activities from which an important part is discharged into the environment. China could be the most affected country by the resulting pollution. However, their ecotoxicological dimension is still less addressed and understood. This study aimed to analyze the toxic effect of sodium isobutyl xanthate (SIBX) and sodium isopropyl xanthate (SIPX) to soil microbial richness and activity and to make a comparison between the two compounds in regard to their effects on soil microbial and enzymes activities. Different methods, including microcalorimetry, viable cell counts, cell density, and catalase and fluorescein diacetate (FDA) hydrololase activities measurement, were applied. The two chemicals exhibited a significant inhibitory effect (P < 0.05 or P < 0.01) to all parameters, SIPX being more adverse than SIBX. As the doses of SIBX and SIPX increased from 5 to 300 μg g -1 soil, their inhibitory ratio ranged from 4.84 to 45.16 % and from 16.13 to 69.68 %, respectively. All parameters fluctuated with the incubation time (10-day period). FDA hydrolysis was more directly affected but was relatively more resilient than catalase activity. Potential changes of those chemicals in the experimental media and complementarity between experimental techniques were justified.

  4. Microbial activity of soil with sulfentrazone associated with phytoremediator species and inoculation with a bacterial consortium

    Directory of Open Access Journals (Sweden)

    Christiane Augusta Diniz Melo

    Full Text Available ABSTRACT Phytostimulation plays a key role in the process of rhizodegradation of herbicides in soil. Additionally, bio-enhancement associated with phytoremediation may increase the efficiency of the decontamination process of soils with herbicides. Therefore, the objective of this study was to evaluate the biomass and microbial activity of soil contaminated with sulfentrazone and cultivated with phytoremediator species plus a bacterial consortium. The experiment was conducted in a greenhouse, carried out with a 2 × 4 × 4 completely randomized factorial design with 4 replications. The first factor consisted of the presence or absence of bio-enhancement with a bacterial consortium composed of Pseudomonas bacteria; the second factor consisted of a monoculture or mixed cultivation of 2 phytoremediator species Canavalia ensiformis and Helianthus annuus, besides the absence of cultivation; the third factor was made up by the bio-remediation time (25, 45, 65, and 85 days after thinning. Uncultivated soils displayed low values of microbial biomass carbon and microbial quotient as well as high values of metabolic quotient throughout the bio-remediation time, indicating the importance of cultivating phytoremediator species for the stimulation of soil microbiota. Bio-enhancement with the bacterial consortium, in general, promoted an increase in the microbial biomass and activity of soil contaminated with sulfentrazone. In the presence of the bacterial consortium, Canavalia ensiformis stimulated a greater activity of associated microbiota and supported a higher microbial biomass. Phytoremediation associated with microbial bio-enhancement are thus promising techniques for the bio-remediation for soils contaminated with sulfentrazone. This technique enhances the biomass and activity of soil microorganisms.

  5. Site- and horizon-specific patterns of microbial community structure and enzyme activities in permafrost-affected soils of Greenland

    DEFF Research Database (Denmark)

    Gittel, Antje; Barta, Jiri; Kohoutova, Iva

    2014-01-01

    topsoils”), resulting from a decrease in fungal abundance compared to recent (“unburied”) topsoils. Actinobacteria (in particular Intrasporangiaceae) accounted for a major fraction of the microbial community in buried topsoils, but were only of minor abundance in all other soil horizons. It was indicated...... that the distribution pattern of Actinobacteria and a variety of other bacterial classes was related to the activity of phenol oxidases and peroxidases supporting the hypothesis that bacteria might resume the role of fungi in oxidative enzyme production and degradation of phenolic and other complex substrates...

  6. Ethanol production from biomass: technology and commercialisation status

    Energy Technology Data Exchange (ETDEWEB)

    Mielenz, J.R.

    2001-06-01

    Owing to technical improvements in the processes used to produce ethanol from biomass, construction of at least two waste-to-ethanol production plants in the United States is expected to start this year. Although there are a number of robust fermentation microorganisms available, initial pretreatment of the biomass and costly cellulase enzymes remain critical targets for process and cost improvements. A highly efficient, very low-acid pretreatment process is approaching pilot testing, while research on cellulases for ethanol production is expanding at both enzyme and organism level. (Author)

  7. Novel Enzymes for Targeted Hydrolysis of Algal Cell Walls

    DEFF Research Database (Denmark)

    Schultz-Johansen, Mikkel

    Seaweeds, also known as macroalgae, constitute a rich source of valuable biomolecules which have a potential industrial application in food and pharma products. The use of enzymes can optimize the extraction and separation of these molecules from the seaweed biomass, but most commercial enzymes...... are incapable of breaking the complex polysaccharides found in seaweed cell walls. Therefore, new enzymes are needed for degradation of seaweed biomass. Bacteria that colonize the surfaces of seaweed secrete enzymes that allow them to degrade and utilize seaweed polysaccharides as energy. In addition, sea...... degradation. In addition, three carrageenases were characterised; one as a GH16 κ-carrageenase whereas the other two belong to a new GH16 subfamily of enzymes that degrade furcellaran (κ/β-carrageenan). From metagenome sequence data three putative GH107 fucanases were identified and characterized...

  8. Microbial production host selection for converting second-generation feedstocks into bioproducts

    NARCIS (Netherlands)

    Rumbold, K.; Buijsen, H.J.J. van; Overkamp, K.M.; Groenestijn, J.W. van; Punt, P.J.; Werf, M.J.V.D.

    2009-01-01

    Increasingly lignocellulosic biomass hydrolysates are used as the feedstock for industrial fermentations. These biomass hydrolysates are complex mixtures of different fermentable sugars, but also inhibitors and salts that affect the performance of the microbial production host. The performance of

  9. The biofilm ecology of microbial biofouling, biocide resistance and corrosion

    Energy Technology Data Exchange (ETDEWEB)

    White, D.C. [Univ. of Tennessee, Knoxville, TN (United States). Center for Environmental Biotechnology]|[Oak Ridge National Lab., TN (United States). Environmental Science Div.; Kirkegaard, R.D.; Palmer, R.J. Jr.; Flemming, C.A.; Chen, G.; Leung, K.T.; Phiefer, C.B. [Univ. of Tennessee, Knoxville, TN (United States). Center for Environmental Biotechnology; Arrage, A.A. [Univ. of Tennessee, Knoxville, TN (United States). Center for Environmental Biotechnology]|[Microbial Insights, Inc., Rockford, TN (United States)

    1997-06-01

    In biotechnological or bioremediation processes it is often the aim to promote biofilm formation, and maintain active, high density biomass. In other situations, biofouling can seriously restrict effective heat transport, membrane processes, and potentate macrofouling with loss of transportation efficiency. In biotechnological or bioremediation processes it is often the aim to promote biofilm formation, and maintain active, high density biomass. In other situations, biofouling can seriously restrict effective heat transport, membrane processes, and potentate macrofouling with loss of transportation efficiency. Heterogeneous distribution of microbes and/or their metabolic activity can promote microbially influenced corrosion (MIC) which is a multibillion dollar problem. Consequently, it is important that biofilm microbial ecology be understood so it can be manipulated rationally. It is usually simple to select organisms that form biofilms by flowing a considerably dilute media over a substratum, and propagating the organisms that attach. To examine the biofilm most expeditiously, the biomass accumulation, desquamation, and metabolic activities need to be monitored on-line and non-destructively. This on-line monitoring becomes even more valuable if the activities can be locally mapped in time and space within the biofilm. Herein the authors describe quantitative measures of microbial biofouling, the ecology of pathogens in drinking water distributions systems, and localization of microbial biofilms and activities with localized MIC.

  10. Temporal variability of available P, microbial P and some ...

    African Journals Online (AJOL)

    Temporal variability of available P, microbial P and some phosphomonoesterase activities in a sewage sludge treated soil: The effect of soil water potential. ... African Journal of Biotechnology ... The objective of this study was to test the effects of water potential on soil available P, microbial biomass P(MBP) and some

  11. Marine Metagenome as A Resource for Novel Enzymes

    KAUST Repository

    Alma’abadi, Amani D.

    2015-11-10

    More than 99% of identified prokaryotes, including many from the marine environment, cannot be cultured in the laboratory. This lack of capability restricts our knowledge of microbial genetics and community ecology. Metagenomics, the culture-independent cloning of environmental DNAs that are isolated directly from an environmental sample, has already provided a wealth of information about the uncultured microbial world. It has also facilitated the discovery of novel biocatalysts by allowing researchers to probe directly into a huge diversity of enzymes within natural microbial communities. Recent advances in these studies have led to great interest in recruiting microbial enzymes for the development of environmentally-friendly industry. Although the metagenomics approach has many limitations, it is expected to provide not only scientific insights but also economic benefits, especially in industry. This review highlights the importance of metagenomics in mining microbial lipases, as an example, by using high-throughput techniques. In addition, we discuss challenges in the metagenomics as an important part of bioinformatics analysis in big data.

  12. Impact of drying-rewetting events on the response of soil microbial functions to dairyfibre and Miscanthus biochars

    Science.gov (United States)

    Bonnett, Sam; Vink, Stefanie; Baker, Kate; Saghir, Muhammad; Hornung, Andreas

    2014-05-01

    Biochar application has been shown to positively affect soil microbial functions such as reducing greenhouse gas emissions, increasing water/nutrient availability and increasing crop yields in tropical regions (Lehmann & Joseph, 2009). Understanding the dynamics of biochar application to soil microbial processes is critical for ensuring that soil quality, integrity and sustainability of the soil sub-system are maintained for crop growth. The aim of this British Ecological Society (BES) funded study was to examine the effect of two types of biochar on soil physicochemistry, GHG production, soil enzyme activities and microbial biomass in typical agricultural soil types and whether the effects were altered by drying, rewetting and flooding events. Miscanthus and dairyfibre (a mixture of straw and manure) feedstocks from Harper Adams University were pyrolyzed by Aston University at 450 °C using 100 kg/hr pyroformer technology. Two sieved soil types (sandy loam and clay loam) were mixed with dry biochar to produce 2 and 10 % w/w treatments for comparison with controls and maintained at 15 °C in temperature controlled incubators. At 0, 22, 44, 80, 101, and 114 days, soil was collected for determination of heterotrophic respiration, and microbial biomass by substrate-induced respiration (SIR), by gas headspace incubation and analysis of carbon dioxide (CO2) and nitrous oxide (N2O) by gas chromatography. Soil was sampled for the determination of water-extractable carbon, pH, and extracellular enzyme activities. Soil samples were maintained at field gravimetric water content between 0 and 44 days; air dried between 44 and 80 days; rewetted between 80 and 101 days; and flooded between 101 to 114 days. Results showed that the impact of biochar on soil microbial processes was dependent on biochar type and soil type, the level of biochar application and changes in soil moisture. Biochar affected soil pH particularly within the dairyfibre treatments, potentially due to the

  13. Microbial community dynamics and transformation of vascular plant detritus in two wetland ecosystems

    International Nuclear Information System (INIS)

    Moran, M.A.

    1987-01-01

    The microbial ecology of two wetland ecosystems in southeastern Georgia, USA, was studied with respect to microbial community dynamics and microbially-mediated transformations of vascular plant detritus. In the Okefenokee Swamp, biomass of microorganisms in the water column and sediments was generally lower in winter months and higher during spring and summer. Biomass and activity (measured as 14 C-lignocellulose mineralization) differed significantly among five habitats within the Okefenokee, and also among locations within each habitat. Significant heterogeneity in the structure of Okefenokee microbial communities was found at scales from 30 cm to 150 m. In field and laboratory studies of vascular plant decomposition in the Okefenokee and a salt marsh on Sapelo Island, the mathematical model which best describes decomposition kinetics is the decaying coefficient model

  14. Conversion of Wastes into Bioelectricity and Chemicals by Using Microbial Electrochemical Technologies

    KAUST Repository

    Logan, B. E.; Rabaey, K.

    2012-01-01

    Waste biomass is a cheap and relatively abundant source of electrons for microbes capable of producing electrical current outside the cell. Rapidly developing microbial electrochemical technologies, such as microbial fuel cells, are part of a

  15. Activity-based protein profiling of secreted cellulolytic enzyme activity dynamics in Trichoderma reesei QM6a, NG14, and RUT-C30

    Energy Technology Data Exchange (ETDEWEB)

    Anderson, Lindsey N.; Culley, David E.; Hofstad, Beth A.; Chauvigne-Hines, Lacie M.; Zink, Erika M.; Purvine, Samuel O.; Smith, Richard D.; Callister, Stephen J.; Magnuson, Jon M.; Wright, Aaron T.

    2013-12-01

    Development of alternative, non-petroleum based sources of bioenergy that can be applied in the short-term find great promise in the use of highly abundant and renewable lignocellulosic plant biomass.1 This material obtained from different feedstocks, such as forest litter or agricultural residues, can yield liquid fuels and other chemical products through biorefinery processes.2 Biofuels are obtained from lignocellulosic materials by chemical pretreatment of the biomass, followed by enzymatic decomposition of cellulosic and hemicellulosic compounds into soluble sugars that are converted to desired chemical products via microbial metabolism and fermentation.3, 4 To release soluble sugars from polymeric cellulose multiple enzymes are required, including endoglucanase, exoglucanase, and β-glucosidase.5, 6 However, the enzymatic hydrolysis of cellulose into soluble sugars remains a significant limiting factor to the efficient and economically viable utilization of lignocellulosic biomass for transport fuels.7, 8 The primary industrial source of cellulose and hemicellulases is the mesophilic soft-rot fungus Trichoderma reesei,9 having widespread applications in food, feed, textile, pulp, and paper industries.10 The genome encodes 200 glycoside hydrolases, including 10 cellulolytic and 16 hemicellulolytic enzymes.11 The hypercellulolytic catabolite derepressed strain RUT-C30 was obtained through a three-step UV and chemical mutagenesis of the original T. reesei strain QM6a,12, 13 in which strains M7 and NG14 were intermediate, having higher cellulolytic activity than the parent strain but less activity and higher catabolite repression than RUT-C30.14 Numerous methods have been employed to optimize the secreted enzyme cocktail of T. reesei including cultivation conditions, operational parameters, and mutagenesis.3 However, creating an optimal and economical enzyme mixture for production-scale biofuels synthesis may take thousands of experiments to identify.

  16. A theoretical reassessment of microbial maintenance and implications for microbial ecology modeling.

    Science.gov (United States)

    Wang, Gangsheng; Post, Wilfred M

    2012-09-01

    We attempted to reconcile three microbial maintenance models (Herbert, Pirt, and Compromise) through a theoretical reassessment. We provided a rigorous proof that the true growth yield coefficient (Y(G)) is the ratio of the specific maintenance rate (a in Herbert) to the maintenance coefficient (m in Pirt). Other findings from this study include: (1) the Compromise model is identical to the Herbert for computing microbial growth and substrate consumption, but it expresses the dependence of maintenance on both microbial biomass and substrate; (2) the maximum specific growth rate in the Herbert (μ(max,H)) is higher than those in the other two models (μ(max,P) and μ(max,C)), and the difference is the physiological maintenance factor (m(q) = a); and (3) the overall maintenance coefficient (m(T)) is more sensitive to m(q) than to the specific growth rate (μ(G)) and Y(G). Our critical reassessment of microbial maintenance provides a new approach for quantifying some important components in soil microbial ecology models. © This article is a US government work and is in the public domain in the USA.

  17. Microbial deterioration of Mayan stone buildings at Uxmal, Yucatan, Mexico

    International Nuclear Information System (INIS)

    Ortega-Morales, O.; Guezennec, J.; Hernandez D, G.; Jozsa, P.; Sand, W.; Crassous, P.

    1998-01-01

    The microbial communities associated to Uxmal Mayan monuments (Yucatan, Mexico) and their role in stone deterioration were preliminary characterized by chemical, biochemical, microbiological, microscopical and surface analysis methods under two climatic seasons (1997). The organic matter and organic carbon and nitrogen were in the range of those reported for other stone buildings, indicating that oligo trophic conditions prevail at Uxmal. Quantitative differences in microbial biomass was higher at indoor section were the organic matter content was the highest and micro-environmental conditions (availability of water and protection to direct sunlight) are more suitable for microbial growth. The microbiological analysis underestimated the microbial biomass, as revealed by biochemical approaches. Nitrate and nitrite-oxidizing, metilotrophic and heterotrophic bacteria and fungi were detected in most surfaces. The heterotrophic bacteria were the most abundant microbial group (microbiological data). However, the chlorophyll profiles and Scanning Electron Microscopy showed that the microalgae are the most abundant colonizers in Uxmal stone buildings. EDAX analysis showed that the most surfaces were covered by an organic layer (cells and exo polymers). Gypsum was found in few samples. The large photo trophic biomass seems to play a role in stone bio deterioration by supporting growth of heterotrophic microorganisms (bacteria and fungi) which are known to produce organic acids leading to calcite dissolution and cations chelation. Further studies are being carried out in order to determine the role of exo polysaccharides which are thought to play a role in chemical degradation of limestone substrates in Uxmal. (Author)

  18. Genome-Enabled Modeling of Biogeochemical Processes Predicts Metabolic Dependencies that Connect the Relative Fitness of Microbial Functional Guilds

    Science.gov (United States)

    Brodie, E.; King, E.; Molins, S.; Karaoz, U.; Steefel, C. I.; Banfield, J. F.; Beller, H. R.; Anantharaman, K.; Ligocki, T. J.; Trebotich, D.

    2015-12-01

    Pore-scale processes mediated by microorganisms underlie a range of critical ecosystem services, regulating carbon stability, nutrient flux, and the purification of water. Advances in cultivation-independent approaches now provide us with the ability to reconstruct thousands of genomes from microbial populations from which functional roles may be assigned. With this capability to reveal microbial metabolic potential, the next step is to put these microbes back where they belong to interact with their natural environment, i.e. the pore scale. At this scale, microorganisms communicate, cooperate and compete across their fitness landscapes with communities emerging that feedback on the physical and chemical properties of their environment, ultimately altering the fitness landscape and selecting for new microbial communities with new properties and so on. We have developed a trait-based model of microbial activity that simulates coupled functional guilds that are parameterized with unique combinations of traits that govern fitness under dynamic conditions. Using a reactive transport framework, we simulate the thermodynamics of coupled electron donor-acceptor reactions to predict energy available for cellular maintenance, respiration, biomass development, and enzyme production. From metagenomics, we directly estimate some trait values related to growth and identify the linkage of key traits associated with respiration and fermentation, macromolecule depolymerizing enzymes, and other key functions such as nitrogen fixation. Our simulations were carried out to explore abiotic controls on community emergence such as seasonally fluctuating water table regimes across floodplain organic matter hotspots. Simulations and metagenomic/metatranscriptomic observations highlighted the many dependencies connecting the relative fitness of functional guilds and the importance of chemolithoautotrophic lifestyles. Using an X-Ray microCT-derived soil microaggregate physical model combined

  19. Contemporary enzyme based technologies for bioremediation: A review.

    Science.gov (United States)

    Sharma, Babita; Dangi, Arun Kumar; Shukla, Pratyoosh

    2018-03-15

    The persistent disposal of xenobiotic compounds like insecticides, pesticides, fertilizers, plastics and other hydrocarbon containing substances is the major source of environmental pollution which needs to be eliminated. Many contemporary remediation methods such as physical, chemical and biological are currently being used, but they are not sufficient to clean the environment. The enzyme based bioremediation is an easy, quick, eco-friendly and socially acceptable approach used for the bioremediation of these recalcitrant xenobiotic compounds from the natural environment. Several microbial enzymes with bioremediation capability have been isolated and characterized from different natural sources, but less production of such enzymes is a limiting their further exploitation. The genetic engineering approach has the potential to get large amount of recombinant enzymes. Along with this, enzyme immobilization techniques can boost the half-life, stability and activity of enzymes at a significant level. Recently, nanozymes may offer the potential bioremediation ability towards a broad range of pollutants. In the present review, we have described a brief overview of the microbial enzymes, different enzymes techniques (genetic engineering and immobilization of enzymes) and nanozymes involved in bioremediation of toxic, carcinogenic and hazardous environmental pollutants. Copyright © 2018 Elsevier Ltd. All rights reserved.

  20. In vitro bioconversion of chitin to pyruvate with thermophilic enzymes.

    Science.gov (United States)

    Honda, Kohsuke; Kimura, Keisuke; Ninh, Pham Huynh; Taniguchi, Hironori; Okano, Kenji; Ohtake, Hisao

    2017-09-01

    Chitin is the second most abundant organic compound on the planet and thus has been regarded as an alternative resource to petroleum feedstocks. One of the key challenges in the biological conversion of biomass-derived polysaccharides, such as cellulose and chitin, is to close the gap between optimum temperatures for enzymatic saccharification and microbial fermentation and to implement them in a single bioreactor. To address this issue, in the present study, we aimed to perform an in vitro, one-pot bioconversion of chitin to pyruvate, which is a precursor of a wide range of useful metabolites. Twelve thermophilic enzymes, including that for NAD + regeneration, were heterologously produced in Escherichia coli and semi-purified by heat treatment of the crude extract of recombinant cells. When the experimentally decided concentrations of enzymes were incubated with 0.5 mg mL -1 colloidal chitin (equivalent to 2.5 mM N-acetylglucosamine unit) and an adequate set of cofactors at 70°C, 0.62 mM pyruvate was produced in 5 h. Despite the use of a cofactor-balanced pathway, determination of the pool sizes of cofactors showed a rapid decrease in ATP concentration, most probably due to the thermally stable ATP-degrading enzyme(s) derived from the host cell. Integration of an additional enzyme set of thermophilic adenylate kinase and polyphosphate kinase led to the deceleration of ATP degradation, and the final product titer was improved to 2.1 mM. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  1. Effects of Recurring Droughts on Extracellular Enzyme Activity in Mountain Grassland

    Science.gov (United States)

    Fuchslueger, L.; Bahn, M.; Kienzl, S.; Hofhansl, F.; Schnecker, J.; Richter, A.

    2015-12-01

    Water availability is a key factor for biogeochemical processes and determines microbial activity and functioning, and thereby organic matter decomposition in soils by affecting the osmotic potential, soil pore connectivity, substrate diffusion and nutrient availability. Low water availability during drought periods therefore directly affects microbial activity. Recurring drought periods likely induce shifts in microbial structure that might be reflected in altered responses of microbial turnover of organic matter by extracellular enzymes. To study this we measured a set of potential extracellular enzyme activity rates (cellobiohydrolase CBH; leucine-amino-peptidase LAP; phosphatase PHOS; phenoloxidase POX), in grassland soils that were exposed to extreme experimental droughts during the growing seasons of up to five subsequent years. During the first drought period after eight weeks of rain exclusion all measured potential enzyme activities were significantly decreased. In parallel, soil extractable organic carbon and nitrogen concentrations increased and microbial community structure, determined by phospholipid fatty acid analysis, changed. In soils that were exposed to two and three drought periods only PHOS decreased. After four years of drought again CBH, PHOS and POX decreased, while LAP was unaffected; after five years of drought PHOS and POX decreased and CBH and LAP remained stable. Thus, our results suggest that recurring extreme drought events can cause different responses of extracellular enzyme activities and that the responses change over time. We will discuss whether and to what degree these changes were related to shifts in microbial community composition. However, independent of whether a solitary or a recurrent drought was imposed, in cases when enzyme activity rates were altered during drought, they quickly recovered after rewetting. Overall, our data suggest that microbial functioning in mountain grassland is sensitive to drought, but highly

  2. Extracellular enzymes facilitate electron uptake in biocorrosion and bioelectrosynthesis.

    Science.gov (United States)

    Deutzmann, Jörg S; Sahin, Merve; Spormann, Alfred M

    2015-04-21

    Direct, mediator-free transfer of electrons between a microbial cell and a solid phase in its surrounding environment has been suggested to be a widespread and ecologically significant process. The high rates of microbial electron uptake observed during microbially influenced corrosion of iron [Fe(0)] and during microbial electrosynthesis have been considered support for a direct electron uptake in these microbial processes. However, the underlying molecular mechanisms of direct electron uptake are unknown. We investigated the electron uptake characteristics of the Fe(0)-corroding and electromethanogenic archaeon Methanococcus maripaludis and discovered that free, surface-associated redox enzymes, such as hydrogenases and presumably formate dehydrogenases, are sufficient to mediate an apparent direct electron uptake. In genetic and biochemical experiments, we showed that these enzymes, which are released from cells during routine culturing, catalyze the formation of H2 or formate when sorbed to an appropriate redox-active surface. These low-molecular-weight products are rapidly consumed by M. maripaludis cells when present, thereby preventing their accumulation to any appreciable or even detectable level. Rates of H2 and formate formation by cell-free spent culture medium were sufficient to explain the observed rates of methane formation from Fe(0) and cathode-derived electrons by wild-type M. maripaludis as well as by a mutant strain carrying deletions in all catabolic hydrogenases. Our data collectively show that cell-derived free enzymes can mimic direct extracellular electron transfer during Fe(0) corrosion and microbial electrosynthesis and may represent an ecologically important but so far overlooked mechanism in biological electron transfer. The intriguing trait of some microbial organisms to engage in direct electron transfer is thought to be widespread in nature. Consequently, direct uptake of electrons into microbial cells from solid surfaces is assumed

  3. Microbial activity and community composition during bioremediation of diesel-oil-contaminated soil: effects of hydrocarbon concentration, fertilizers, and incubation time.

    Science.gov (United States)

    Margesin, Rosa; Hämmerle, Marion; Tscherko, Dagmar

    2007-02-01

    We investigated the influence of three factors-diesel oil concentration [2500, 5000, 10,000, 20,000 mg total petroleum hydrocarbons (TPH) kg(-1) soil], biostimulation (unfertilized, inorganic fertilization with NPK nutrients, or oleophilic fertilization with Inipol EAP22), and incubation time-on hydrocarbon removal, enzyme activity (lipase), and microbial community structure [phospholipid fatty acids (PLFA)] in a laboratory soil bioremediation treatment. Fertilization enhanced TPH removal and lipase activity significantly (P 0.05). Microbial communities, as assessed by PLFA patterns, were primarily influenced by the TPH content, followed by fertilization, and the interaction of these two factors, whereas incubation time was of minor importance. This was demonstrated by three-factorial analysis of variance and multidimensional scaling analysis. Low TPH content had no significant effect on soil microbial community, independent of the treatment. High TPH content generally resulted in increased PLFA concentrations, whereby a significant increase in microbial biomass with time was only observed with inorganic fertilization, whereas oleophilic fertilization (Inipol EAP22) tended to inhibit microbial activity and to reduce PLFA contents with time. Among bacteria, PLFA indicative of the Gram-negative population were significantly (P diesel oil and fertilized with NPK after 21-38 days of incubation at 20 degrees C. The Gram-positive population was not significantly influenced by TPH content or biostimulation treatment.

  4. Extracellular enzyme kinetics scale with resource availability

    Science.gov (United States)

    Sinsabaugh, Robert L.; Belnap, Jayne; Findlay, Stuart G.; Follstad Shah, Jennifer J.; Hill, Brian H.; Kuehn, Kevin A.; Kuske, Cheryl; Litvak, Marcy E.; Martinez, Noelle G.; Moorhead, Daryl L.; Warnock, Daniel D.

    2014-01-01

    Microbial community metabolism relies on external digestion, mediated by extracellular enzymes that break down complex organic matter into molecules small enough for cells to assimilate. We analyzed the kinetics of 40 extracellular enzymes that mediate the degradation and assimilation of carbon, nitrogen and phosphorus by diverse aquatic and terrestrial microbial communities (1160 cases). Regression analyses were conducted by habitat (aquatic and terrestrial), enzyme class (hydrolases and oxidoreductases) and assay methodology (low affinity and high affinity substrates) to relate potential reaction rates to substrate availability. Across enzyme classes and habitats, the scaling relationships between apparent Vmax and apparent Km followed similar power laws with exponents of 0.44 to 0.67. These exponents, called elasticities, were not statistically distinct from a central value of 0.50, which occurs when the Km of an enzyme equals substrate concentration, a condition optimal for maintenance of steady state. We also conducted an ecosystem scale analysis of ten extracellular hydrolase activities in relation to soil and sediment organic carbon (2,000–5,000 cases/enzyme) that yielded elasticities near 1.0 (0.9 ± 0.2, n = 36). At the metabolomic scale, the elasticity of extracellular enzymatic reactions is the proportionality constant that connects the C:N:P stoichiometries of organic matter and ecoenzymatic activities. At the ecosystem scale, the elasticity of extracellular enzymatic reactions shows that organic matter ultimately limits effective enzyme binding sites. Our findings suggest that one mechanism by which microbial communities maintain homeostasis is regulating extracellular enzyme expression to optimize the short-term responsiveness of substrate acquisition. The analyses also show that, like elemental stoichiometry, the fundamental attributes of enzymatic reactions can be extrapolated from biochemical to community and ecosystem scales.

  5. Incorporating microbial dormancy dynamics into soil decomposition models to improve quantification of soil carbon dynamics of northern temperate forests

    Science.gov (United States)

    He, Yujie; Yang, Jinyan; Zhuang, Qianlai; Harden, Jennifer W.; McGuire, A. David; Liu, Yaling; Wang, Gangsheng; Gu, Lianhong

    2015-01-01

    Soil carbon dynamics of terrestrial ecosystems play a significant role in the global carbon cycle. Microbial-based decomposition models have seen much growth recently for quantifying this role, yet dormancy as a common strategy used by microorganisms has not usually been represented and tested in these models against field observations. Here we developed an explicit microbial-enzyme decomposition model and examined model performance with and without representation of microbial dormancy at six temperate forest sites of different forest types. We then extrapolated the model to global temperate forest ecosystems to investigate biogeochemical controls on soil heterotrophic respiration and microbial dormancy dynamics at different temporal-spatial scales. The dormancy model consistently produced better match with field-observed heterotrophic soil CO2 efflux (RH) than the no dormancy model. Our regional modeling results further indicated that models with dormancy were able to produce more realistic magnitude of microbial biomass (analysis showed that soil organic carbon content was the dominating factor (correlation coefficient = 0.4–0.6) in the simulated spatial pattern of soil RHwith both models. In contrast to strong temporal and local controls of soil temperature and moisture on microbial dormancy, our modeling results showed that soil carbon-to-nitrogen ratio (C:N) was a major regulating factor at regional scales (correlation coefficient = −0.43 to −0.58), indicating scale-dependent biogeochemical controls on microbial dynamics. Our findings suggest that incorporating microbial dormancy could improve the realism of microbial-based decomposition models and enhance the integration of soil experiments and mechanistically based modeling.

  6. Production, purification and characterization of fibrinolytic enzyme from Serratia sp. KG-2-1 using optimized media.

    Science.gov (United States)

    Taneja, Kapila; Bajaj, Bijender Kumar; Kumar, Sandeep; Dilbaghi, Neeraj

    2017-07-01

    Intravascular thrombosis is one of the major causes of variety of cardiovascular disorders leading to high mortality worldwide. Fibrinolytic enzymes from microbial sources possess ability to dissolve these clots and help to circumvent these problems in more efficient and safer way. In the present study, fibrinolytic protease with higher fibrinolytic activity than plasmin was obtained from Serratia sp. KG-2-1 isolated from garbage dump soil. Response surface methodology was used to study the interactive effect of concentration of maltose, yeast extract + peptone (1:1), incubation time, and pH on enzyme production and biomass. Maximum enzyme production was achieved at 33 °C after 24 h at neutral pH in media containing 1.5% Maltose, 4.0% yeast extract + peptone and other trace elements resulting in 1.82 folds increased production. The enzyme was purified from crude extract using ammonium sulfate precipitation and DEAE-Sephadex chromatography resulting in 12.9 fold purification with 14.9% yield. The purified enzyme belongs to metalloprotease class and had optimal activity in conditions similar to physiological environment with temperature optima of 40 °C and pH optima of 8. The enzyme was found to be stable in various solvents and its activity was enhanced in presence of Na + , K + , Ba 2+ , Cu 2+ , Mn 2+ , Hg 2+ but inhibited by Ca 2+ and Fe 3+ . Hence, the obtained enzyme may be used as potential therapeutic agent in combating various thrombolytic disorders.

  7. Influence of a direct-fed microbial and xylanase enzyme on the dietary energy uptake efficiency and performance of broiler chickens.

    Science.gov (United States)

    Murugesan, Ganapathi Raj; Persia, Michael E

    2015-09-01

    Efficacy of a multi-strain direct-fed microbial product (PoultryStar(®) ME; PS) and a xylanase enzyme product on the dietary energy utilization efficiency and resulting performance in broiler chickens was evaluated. Apart from performance parameters, cecal and serum metabolites and activities of hepatic enzymes involved in energy metabolism were also determined. Ross 308 chicks were fed one of four experimental diets [control (CON), CON + PS, CON + xylanase and CON + PS + xylanase] using a 2 × 2 factorial arrangement from 1-21 days of age. Cecal proportions of propionate and butyrate, as well as total short-chain fatty acid concentration were increased (P energy uptake and hepatic energy retention. The combination additively increased the FCR, suggesting involvement of synergistic modes of actions. © 2014 Society of Chemical Industry.

  8. Strategies for microbial synthesis of high-value phytochemicals

    Science.gov (United States)

    Li, Sijin; Li, Yanran; Smolke, Christina D.

    2018-03-01

    Phytochemicals are of great pharmaceutical and agricultural importance, but often exhibit low abundance in nature. Recent demonstrations of industrial-scale production of phytochemicals in yeast have shown that microbial production of these high-value chemicals is a promising alternative to sourcing these molecules from native plant hosts. However, a number of challenges remain in the broader application of this approach, including the limited knowledge of plant secondary metabolism and the inefficient reconstitution of plant metabolic pathways in microbial hosts. In this Review, we discuss recent strategies to achieve microbial biosynthesis of complex phytochemicals, including strategies to: (1) reconstruct plant biosynthetic pathways that have not been fully elucidated by mining enzymes from native and non-native hosts or by enzyme engineering; (2) enhance plant enzyme activity, specifically cytochrome P450 activity, by improving efficiency, selectivity, expression or electron transfer; and (3) enhance overall reaction efficiency of multi-enzyme pathways by dynamic control, compartmentalization or optimization with the host's metabolism. We also highlight remaining challenges to — and future opportunities of — this approach.

  9. Efficient plant biomass degradation by thermophilic fungus Myceliophthora heterothallica.

    Science.gov (United States)

    van den Brink, Joost; van Muiswinkel, Gonny C J; Theelen, Bart; Hinz, Sandra W A; de Vries, Ronald P

    2013-02-01

    Rapid and efficient enzymatic degradation of plant biomass into fermentable sugars is a major challenge for the sustainable production of biochemicals and biofuels. Enzymes that are more thermostable (up to 70°C) use shorter reaction times for the complete saccharification of plant polysaccharides compared to hydrolytic enzymes of mesophilic fungi such as Trichoderma and Aspergillus species. The genus Myceliophthora contains four thermophilic fungi producing industrially relevant thermostable enzymes. Within this genus, isolates belonging to M. heterothallica were recently separated from the well-described species M. thermophila. We evaluate here the potential of M. heterothallica isolates to produce efficient enzyme mixtures for biomass degradation. Compared to the other thermophilic Myceliophthora species, isolates belonging to M. heterothallica and M. thermophila grew faster on pretreated spruce, wheat straw, and giant reed. According to their protein profiles and in vitro assays after growth on wheat straw, (hemi-)cellulolytic activities differed strongly between M. thermophila and M. heterothallica isolates. Compared to M. thermophila, M. heterothallica isolates were better in releasing sugars from mildly pretreated wheat straw (with 5% HCl) with a high content of xylan. The high levels of residual xylobiose revealed that enzyme mixtures of Myceliophthora species lack sufficient β-xylosidase activity. Sexual crossing of two M. heterothallica showed that progenies had a large genetic and physiological diversity. In the future, this will allow further improvement of the plant biomass-degrading enzyme mixtures of M. heterothallica.

  10. Microbial physiology and soil CO2 efflux after 9 years of soil warming in a temperate forest - no indications for thermal adaptations.

    Science.gov (United States)

    Schindlbacher, Andreas; Schnecker, Jörg; Takriti, Mounir; Borken, Werner; Wanek, Wolfgang

    2015-11-01

    Thermal adaptations of soil microorganisms could mitigate or facilitate global warming effects on soil organic matter (SOM) decomposition and soil CO2 efflux. We incubated soil from warmed and control subplots of a forest soil warming experiment to assess whether 9 years of soil warming affected the rates and the temperature sensitivity of the soil CO2 efflux, extracellular enzyme activities, microbial efficiency, and gross N mineralization. Mineral soil (0-10 cm depth) was incubated at temperatures ranging from 3 to 23 °C. No adaptations to long-term warming were observed regarding the heterotrophic soil CO2 efflux (R10 warmed: 2.31 ± 0.15 μmol m(-2)  s(-1) , control: 2.34 ± 0.29 μmol m(-2)  s(-1) ; Q10 warmed: 2.45 ± 0.06, control: 2.45 ± 0.04). Potential enzyme activities increased with incubation temperature, but the temperature sensitivity of the enzymes did not differ between the warmed and the control soils. The ratio of C : N acquiring enzyme activities was significantly higher in the warmed soil. Microbial biomass-specific respiration rates increased with incubation temperature, but the rates and the temperature sensitivity (Q10 warmed: 2.54 ± 0.23, control 2.75 ± 0.17) did not differ between warmed and control soils. Microbial substrate use efficiency (SUE) declined with increasing incubation temperature in both, warmed and control, soils. SUE and its temperature sensitivity (Q10 warmed: 0.84 ± 0.03, control: 0.88 ± 0.01) did not differ between warmed and control soils either. Gross N mineralization was invariant to incubation temperature and was not affected by long-term soil warming. Our results indicate that thermal adaptations of the microbial decomposer community are unlikely to occur in C-rich calcareous temperate forest soils. © 2015 The Authors. Global Change Biology published by John Wiley & Sons Ltd.

  11. Visualization of Enzyme Activities in Earthworm Biopores by In Situ Soil Zymography.

    Science.gov (United States)

    Razavi, Bahar S; Hoang, Duyen; Kuzyakov, Yakov

    2017-01-01

    Earthworms produce biopores with strongly increased microbial and enzyme activities and consequently they form microbial hotspots in soil. In extremely dynamic microhabitats and hotspots such as earthworm biopores, the in situ enzyme activities are a footprint of process rates and complex biotic interactions. The effect of earthworms on enzyme activities inside biopores, relative to earthworm-free soil, can be visualized by in situ soil zymography. Here, we describe the details of the approach and discuss its advantages and limitations. Direct zymography provides high spatial resolution for quantitative images of enzyme activities in biopores.

  12. Epigeic earthworms exert a bottleneck effect on microbial communities through gut associated processes.

    Science.gov (United States)

    Gómez-Brandón, María; Aira, Manuel; Lores, Marta; Domínguez, Jorge

    2011-01-01

    Earthworms play a critical role in organic matter decomposition because of the interactions they establish with microorganisms. The ingestion, digestion, assimilation of organic material in the gut and then casting is the first step in earthworm-microorganism interactions. The current knowledge of these direct effects is still limited for epigeic earthworm species, mainly those living in man-made environments. Here we tested whether and to what extent the earthworm Eisenia andrei is capable of altering the microbiological properties of fresh organic matter through gut associated processes; and if these direct effects are related to the earthworm diet. To address these questions we determined the microbial community structure (phospholipid fatty acid profiles) and microbial activity (fluorescein diacetate hydrolysis) in the earthworm casts derived from three types of animal manure (cow, horse and pig manure), which differed in microbial composition. The passage of the organic material through the gut of E. andrei reduced the total microbial biomass irrespective of the type of manure, and resulted in a decrease in bacterial biomass in all the manures; whilst leaving the fungi unaffected in the egested materials. However, unlike the microbial biomass, no such reduction was detected in the total microbial activity of cast samples derived from the pig manure. Moreover, no differences were found between cast samples derived from the different types of manure with regards to microbial community structure, which provides strong evidence for a bottleneck effect of worm digestion on microbial populations of the original material consumed. Our data reveal that earthworm gut is a major shaper of microbial communities, thereby favouring the existence of a reduced but more active microbial population in the egested materials, which is of great importance to understand how biotic interactions within the decomposer food web influence on nutrient cycling.

  13. Epigeic earthworms exert a bottleneck effect on microbial communities through gut associated processes.

    Directory of Open Access Journals (Sweden)

    María Gómez-Brandón

    Full Text Available BACKGROUND: Earthworms play a critical role in organic matter decomposition because of the interactions they establish with microorganisms. The ingestion, digestion, assimilation of organic material in the gut and then casting is the first step in earthworm-microorganism interactions. The current knowledge of these direct effects is still limited for epigeic earthworm species, mainly those living in man-made environments. Here we tested whether and to what extent the earthworm Eisenia andrei is capable of altering the microbiological properties of fresh organic matter through gut associated processes; and if these direct effects are related to the earthworm diet. METHODOLOGY: To address these questions we determined the microbial community structure (phospholipid fatty acid profiles and microbial activity (fluorescein diacetate hydrolysis in the earthworm casts derived from three types of animal manure (cow, horse and pig manure, which differed in microbial composition. PRINCIPAL FINDINGS: The passage of the organic material through the gut of E. andrei reduced the total microbial biomass irrespective of the type of manure, and resulted in a decrease in bacterial biomass in all the manures; whilst leaving the fungi unaffected in the egested materials. However, unlike the microbial biomass, no such reduction was detected in the total microbial activity of cast samples derived from the pig manure. Moreover, no differences were found between cast samples derived from the different types of manure with regards to microbial community structure, which provides strong evidence for a bottleneck effect of worm digestion on microbial populations of the original material consumed. CONCLUSIONS/SIGNIFICANCE: Our data reveal that earthworm gut is a major shaper of microbial communities, thereby favouring the existence of a reduced but more active microbial population in the egested materials, which is of great importance to understand how biotic interactions

  14. Soil fungal and bacterial biomass determined by epifluorescence microscopy and mycorrhizal spore density in different sugarcane managements

    Directory of Open Access Journals (Sweden)

    Adriana Pereira Aleixo

    2014-04-01

    Full Text Available Crop productivity and sustainability have often been related to soil organic matter and soil microbial biomass, especially because of their role in soil nutrient cycling. This study aimed at measuring fungal and bacterial biomass by epifluorescence microscopy and arbuscular mycorrhizal fungal (AMF spore density in sugarcane (Saccharum officinarum L. fields under different managements. We collected soil samples of sugarcane fields managed with or without burning, with or without mechanized harvest, with or without application of vinasse and from nearby riparian native forest. The soil samples were collected at 10cm depth and storage at 4°C until analysis. Fungal biomass varied from 25 to 37µg C g-1 dry soil and bacterial from 178 to 263µg C g-1 dry soil. The average fungal/bacterial ratio of fields was 0.14. The AMF spore density varied from 9 to 13 spores g-1 dry soil. The different sugarcane managements did not affect AMF spore density. In general, there were no significant changes of microbial biomass with crop management and riparian forest. However, the sum of fungal and bacterial biomass measured by epifluorescence microscopy (i.e. 208-301µg C g-1 dry soil was very close to values of total soil microbial biomass observed in other studies with traditional techniques (e.g. fumigation-extraction. Therefore, determination of fungal/bacterial ratios by epifluorescence microscopy, associated with other parameters, appears to be a promising methodology to understand microbial functionality and nutrient cycling under different soil and crop managements.

  15. Degradation of vinasse in soil under different humidity levels: CO sub 2 liberation, microbial biomass formation and immobilization of added nitrogen. Decomposicao de vinhaca em solo sob diferentes niveis de umidade: liberacao de CO sub 2 , formacao de biomassa microbiana e imobilizacao do nitrogenio adicionado

    Energy Technology Data Exchange (ETDEWEB)

    Minhoni, M T.A. [UNESP, Botucatu, SP (Brazil). Dept. de Defesa Fitossanitaria; Cerri, C C [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil)

    1987-01-01

    Degradation of vinasse added to a sandy Red-Yellow Latosol at the rate of 200m{sup 3}/ha and kept at 40,60 and 80% of the holding capacity (w.h.c.), was studied and compared for liberation of CO{sub 2}, formation of microbial biomass and immobilization of nitrogen added. The CO{sub 2} liberated was evaluated by NaOH retention followed by titration with HCl. The microbial biomass was determined by using gamma radiation as biocide. Nitrogen immobilization was determined using the Kjeldahl method and {sup 15}N enrichment according to Rittemberg's method. Soil moisture, which affected the oxygen level of the soil, had a significant influence in CO{sub 2} liberation, formation of microbial biomassa and nitrogen immobilization. Samples kept under drier conditions (40% w.h.c.) showed initially greater Co{sub 2} liberation. However, at the end of 3 month incubation period, total carbon evolved was similar at all misture levels used, with an average of 3805{mu}g C/g soil. The microbial biomass showed greater formation for the drier samples (40% w.h.c.), reaching a maximum of 519{mu}g C/g soil. Immobilization of the N added showed an increasing initial rate, which was greater with dryness of the soil, followed by stabilization. Nevertheless, at the end of 3 month incubation period, the percentages of immobilization were similar and about 40% of total {sup 15}N irrespective of the soil moisture content. Therefore, the increasing rate of carbon assimilation was not totally acompanied by an increasing immobilization for the N added. The greatest intensity was reached by CO{sub 2} liberation in residue degradation, 2/3 of the carbon having evolved to CO{sub 2} and than 1/3 having been immobilized by the microbial biomass. (author).

  16. Solid/solution Cu fractionations/speciation of a Cu contaminated soil after pilot-scale electrokinetic remediation and their relationships with soil microbial and enzyme activities

    International Nuclear Information System (INIS)

    Wang Quanying; Zhou Dongmei; Cang Long; Li Lianzhen; Wang Peng

    2009-01-01

    The aim of this study was to investigate the detailed metal speciation/fractionations of a Cu contaminated soil before and after electrokinetic remediation as well as their relationships with the soil microbial and enzyme activities. Significant changes in the exchangeable and adsorbed-Cu fractionations occurred after electrokinetic treatment, while labile soil Cu in the solution had a tendency to decrease from the anode to the cathode, and the soil free Cu 2+ ions were mainly accumulated in the sections close to the cathode. The results of regression analyses revealed that both the soil Cu speciation in solution phase and the Cu fractionations in solid phase could play important roles in the changes of the soil microbial and enzyme activities. Our findings suggest that the bioavailability of soil heavy metals and their ecotoxicological effects on the soil biota before and after electroremediation can be better understood in terms of their chemical speciation and fractionations. - The assessment of the roles of soil solution speciation and solid-phase fractionations in metal bioavailability after electrokinetic remediation deserves close attention.

  17. Structural analysis of enzymes used for bioindustry and bioremediation.

    Science.gov (United States)

    Tanokura, Masaru; Miyakawa, Takuya; Guan, Lijun; Hou, Feng

    2015-01-01

    Microbial enzymes have been widely applied in the large-scale, bioindustrial manufacture of food products and pharmaceuticals due to their high substrate specificity and stereoselectivity, and their effectiveness under mild conditions with low environmental burden. At the same time, bioremedial techniques using microbial enzymes have been developed to solve the problem of industrial waste, particularly with respect to persistent chemicals and toxic substances. And finally, structural studies of these enzymes have revealed the mechanistic basis of enzymatic reactions, including the stereoselectivity and binding specificity of substrates and cofactors. The obtained structural insights are useful not only to deepen our understanding of enzymes with potential bioindustrial and/or bioremedial application, but also for the functional improvement of enzymes through rational protein engineering. This review shows the structural bases for various types of enzymatic reactions, including the substrate specificity accompanying cofactor-controlled and kinetic mechanisms.

  18. Effect of a temperature gradient on Sphagnum fallax and its associated living microbial communities: a study under controlled conditions.

    Science.gov (United States)

    Jassey, Vincent E J; Gilbert, Daniel; Binet, Philippe; Toussaint, Marie-Laure; Chiapusio, Geneviève

    2011-03-01

    Microbial communities living in Sphagnum are known to constitute early indicators of ecosystem disturbances, but little is known about their response (including their trophic relationships) to climate change. A microcosm experiment was designed to test the effects of a temperature gradient (15, 20, and 25°C) on microbial communities including different trophic groups (primary producers, decomposers, and unicellular predators) in Sphagnum segments (0-3 cm and 3-6 cm of the capitulum). Relationships between microbial communities and abiotic factors (pH, conductivity, temperature, and polyphenols) were also studied. The density and the biomass of testate amoebae in Sphagnum upper segments increased and their community structure changed in heated treatments. The biomass of testate amoebae was linked to the biomass of bacteria and to the total biomass of other groups added and, thus, suggests that indirect effects on the food web structure occurred. Redundancy analysis revealed that microbial assemblages differed strongly in Sphagnum upper segments along a temperature gradient in relation to abiotic factors. The sensitivity of these assemblages made them interesting indicators of climate change. Phenolic compounds represented an important explicative factor in microbial assemblages and outlined the potential direct and (or) indirect effects of phenolics on microbial communities.

  19. Assimilable organic carbon (AOC) in soil water extracts using Vibrio harveyi BB721 and its implication for microbial biomass.

    Science.gov (United States)

    Ma, Jincai; Ibekwe, A Mark; Wang, Haizhen; Xu, Jianming; Leddy, Menu; Yang, Ching-Hong; Crowley, David E

    2012-01-01

    Assimilable organic carbon (AOC) is commonly used to measure the growth potential of microorganisms in water, but has not yet been investigated for measuring microbial growth potential in soils. In this study, a simple, rapid, and non-growth based assay to determine AOC in soil was developed using a naturally occurring luminous strain Vibrio harveyi BB721 to determine the fraction of low molecular weight organic carbon in soil water extract. Calibration of the assay was achieved by measuring the luminescence intensity of starved V. harveyi BB721 cells in the late exponential phase with a concentration range from 0 to 800 µg l(-1) glucose (equivalent to 0-16.0 mg glucose C kg(-1) soil) with the detection limit of 10 µg l(-1) equivalent to 0.20 mg glucose C kg(-1) soil. Results showed that bioluminescence was proportional to the concentration of glucose added to soil. The luminescence intensity of the cells was highly pH dependent and the optimal pH was about 7.0. The average AOC concentration in 32 soils tested was 2.9±2.2 mg glucose C kg(-1). Our data showed that AOC levels in soil water extracts were significantly correlated (Pgrowth bioluminescence based assay. Understanding the levels of AOC in soil water extract provides new insights into our ability to estimate the most available carbon pool to bacteria in soil that may be easily assimilated into cells for many metabolic processes and suggest possible the links between AOC, microbial regrowth potential, and microbial biomass in soils.

  20. Enhanced conversion of newly-added maize straw to soil microbial biomass C under plastic film mulching and organic manure management

    Science.gov (United States)

    Jin, X.; Filley, T. R.

    2017-12-01

    Management of crop residues using plastic film mulching (PFM) has the potential to improve soil health by accelerating nutrient cycling and facilitating stable C pool production; however, a key aspect of this process—microbial immobilization of residue C—is poorly understood, especially under PFM when combined with different fertilization treatments. A 360-day in situ 13C-tracing technique was used to analyze the contribution and dynamics of microbial biomass C (MBC) to soil organic C (SOC) after 13C-labelled maize straw residue was applied to micro-plot topsoil in a cultivated maize (Zea mays L.) field under 27-year PFM and four fertilization treatments. Over the course of the experiment, MBC content was significantly (P<0.05) higher in treatments of manure (M) and manure plus nitrogen (MN) compared to the no-fertilization (CK) and nitrogen (N) treatments, regardless of PFM. Compared to no PFM controls, PFM enhanced the decomposition of maize straw during summer (Day 60) in the M and MN treatments, exhibiting increases of 93.0% and 28.6% in straw-derived 13C-MBC and 80.4% and 82.9% in 13C-MBC/13C-SOC, respectively. Overall, both PFM and organic manure treatments improved soil fertility through microbe-mediated incorporation of C derived from newly-added maize straw. Our results indicate that microbial growth and activity are affected by the utilization of different C sources and most dramatically during early seasonal transition.