Pnp gene modification for improved xylose utilization in Zymomonas

Science.gov (United States)

Caimi, Perry G G; Qi, Min; Tao, Luan; Viitanen, Paul V; Yang, Jianjun

2014-12-16

The endogenous pnp gene encoding polynucleotide phosphorylase in the Zymomonas genome was identified as a target for modification to provide improved xylose utilizing cells for ethanol production. The cells are in addition genetically modified to have increased expression of ribose-5-phosphate isomerase (RPI) activity, as compared to cells without this genetic modification, and are not limited in xylose isomerase activity in the absence of the pnp modification.

The intra- and extracellular proteome of Aspergillus niger growing on defined medium with xylose or maltose as carbon substrate

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Wissing Josef

2010-04-01

glucoamylase (multiple spots was identified as the most abundant extracellular protein. Surprisingly, the intracellular proteome of A. niger growing on xylose in bioreactor cultures differed more from a culture growing in shake flasks using the same medium than from the bioreactor culture growing on maltose. For example, in shake flask cultures with xylose as carbon source the most abundant intracellular proteins were not the glycolytic and the TCA cycle enzymes and the flavohemoglobin, but CipC, a protein of yet unknown function, superoxide dismutase and an NADPH dependent aldehyde reductase. Moreover, vacuolar proteases accumulated to higher and ER-resident chaperones and foldases to lower levels in shake flask compared to the bioreactor cultures. Conclusions The utilization of xylose or maltose was strongly affecting the composition of the secretome but of minor influence on the composition of the intracellular proteome. On the other hand, differences in culture conditions (pH control versus no pH control, aeration versus no aeration and stirring versus shaking have a profound effect on the intracellular proteome. For example, lower levels of ER-resident chaperones and foldases and higher levels of vacuolar proteases render shake flask conditions less favorable for protein production compared to controlled bioreactor cultures.

Evolutionary Adaptation of Kluyveromyces marxianus NIRE-K3 for Enhanced Xylose Utilization

International Nuclear Information System (INIS)

Sharma, Nilesh Kumar; Behera, Shuvashish; Arora, Richa; Kumar, Sachin

2017-01-01

The evolutionary adaptation was approached on the thermotolerant yeast Kluyveromyces marxianus NIRE-K3 at 45°C on xylose as a sole source of carbon for enhancement of xylose uptake. After 60 cycles, evolved strain K. marxianus NIRE-K3.1 showed comparatively 3.75- and 3.0-fold higher specific growth and xylose uptake rates, respectively, than that of native strain. Moreover, the short lag phase was also observed on adapted strain. During batch fermentation with xylose concentration of 30 g l −1 , K. marxianus NIRE-K3.1 could utilize about 96% of xylose in 72 h and produced 4.67 and 15.7 g l −1 of ethanol and xylitol, respectively, which were 9.72- and 4.63-fold higher than that of native strain. Similarly, specific sugar consumption rate, xylitol, and ethanol yields were 5.07-, 1.15-, and 2.44-fold higher as compared to the native strain, respectively. The results obtained after evolutionary adaptation of K. marxianus NIRE-K3 show the significant improvement in the xylose utilization, ethanol and xylitol yields, and productivities. By understanding the results obtained, the significance of evolutionary adaptation has been rationalized, since the adapted culture could be more stable and could enhance the productivity.

Evolutionary Adaptation of Kluyveromyces marxianus NIRE-K3 for Enhanced Xylose Utilization

Energy Technology Data Exchange (ETDEWEB)

Sharma, Nilesh Kumar [Biochemical Conversion Division, Sardar Swaran Singh National Institute of Bio-Energy, Kapurthala (India); I. K. Gujral Punjab Technical University, Kapurthala (India); Behera, Shuvashish; Arora, Richa; Kumar, Sachin, E-mail: sachin.biotech@gmail.com [Biochemical Conversion Division, Sardar Swaran Singh National Institute of Bio-Energy, Kapurthala (India)

2017-12-12

The evolutionary adaptation was approached on the thermotolerant yeast Kluyveromyces marxianus NIRE-K3 at 45°C on xylose as a sole source of carbon for enhancement of xylose uptake. After 60 cycles, evolved strain K. marxianus NIRE-K3.1 showed comparatively 3.75- and 3.0-fold higher specific growth and xylose uptake rates, respectively, than that of native strain. Moreover, the short lag phase was also observed on adapted strain. During batch fermentation with xylose concentration of 30 g l{sup −1}, K. marxianus NIRE-K3.1 could utilize about 96% of xylose in 72 h and produced 4.67 and 15.7 g l{sup −1} of ethanol and xylitol, respectively, which were 9.72- and 4.63-fold higher than that of native strain. Similarly, specific sugar consumption rate, xylitol, and ethanol yields were 5.07-, 1.15-, and 2.44-fold higher as compared to the native strain, respectively. The results obtained after evolutionary adaptation of K. marxianus NIRE-K3 show the significant improvement in the xylose utilization, ethanol and xylitol yields, and productivities. By understanding the results obtained, the significance of evolutionary adaptation has been rationalized, since the adapted culture could be more stable and could enhance the productivity.

Improved xylose and arabinose utilization by an industrial recombinant Saccharomyces cerevisiae strain using evolutionary engineering

DEFF Research Database (Denmark)

Sanchez, R.G.; Karhumaa, Kaisa; Fonseca, C.

2010-01-01

Background: Cost-effective fermentation of lignocellulosic hydrolysate to ethanol by Saccharomyces cerevisiae requires efficient mixed sugar utilization. Notably, the rate and yield of xylose and arabinose co-fermentation to ethanol must be enhanced. Results: Evolutionary engineering was used...... to improve the simultaneous conversion of xylose and arabinose to ethanol in a recombinant industrial Saccharomyces cerevisiae strain carrying the heterologous genes for xylose and arabinose utilization pathways integrated in the genome. The evolved strain TMB3130 displayed an increased consumption rate...... of our knowledge, this is the first report that characterizes the molecular mechanisms for improved mixed-pentose utilization obtained by evolutionary engineering of a recombinant S. cerevisiae strain. Increased transport of pentoses and increased activities of xylose converting enzymes contributed...

Comparative genomic and transcriptomic analysis revealed genetic characteristics related to solvent formation and xylose utilization in Clostridium acetobutylicum EA 2018

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Wang Shengyue

2011-02-01

Full Text Available Abstract Background Clostridium acetobutylicum, a gram-positive and spore-forming anaerobe, is a major strain for the fermentative production of acetone, butanol and ethanol. But a previously isolated hyper-butanol producing strain C. acetobutylicum EA 2018 does not produce spores and has greater capability of solvent production, especially for butanol, than the type strain C. acetobutylicum ATCC 824. Results Complete genome of C. acetobutylicum EA 2018 was sequenced using Roche 454 pyrosequencing. Genomic comparison with ATCC 824 identified many variations which may contribute to the hyper-butanol producing characteristics in the EA 2018 strain, including a total of 46 deletion sites and 26 insertion sites. In addition, transcriptomic profiling of gene expression in EA 2018 relative to that of ATCC824 revealed expression-level changes of several key genes related to solvent formation. For example, spo0A and adhEII have higher expression level, and most of the acid formation related genes have lower expression level in EA 2018. Interestingly, the results also showed that the variation in CEA_G2622 (CAC2613 in ATCC 824, a putative transcriptional regulator involved in xylose utilization, might accelerate utilization of substrate xylose. Conclusions Comparative analysis of C. acetobutylicum hyper-butanol producing strain EA 2018 and type strain ATCC 824 at both genomic and transcriptomic levels, for the first time, provides molecular-level understanding of non-sporulation, higher solvent production and enhanced xylose utilization in the mutant EA 2018. The information could be valuable for further genetic modification of C. acetobutylicum for more effective butanol production.

NADPH-dependent D-aldose reductases and xylose fermentation in Fusarium oxysporum

DEFF Research Database (Denmark)

Panagiotou, Gianni; Christakopoulos, P.

2004-01-01

for NADPH over NADH. In this study, the influence of aeration and the response to the addition of electron acceptors on xylose fermentation by F. oxysporum were also studied. The batch cultivation of F. oxysporum on xylose was performed under aerobic, anaerobic and oxygen-limited conditions in stirred tank...... conditions (0.3 vvm). When the artificial electron acceptor acetoin was added to an anaerobic batch fermentation of xylose by F. oxysporum, the ethanol yield increased while xylitol excretion was also decreased....

Co-Utilization of Glucose and Xylose for Enhanced Lignocellulosic Ethanol Production with Reverse Membrane Bioreactors

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Mofoluwake M. Ishola

2015-12-01

Full Text Available Integrated permeate channel (IPC flat sheet membranes were examined for use as a reverse membrane bioreactor (rMBR for lignocellulosic ethanol production. The fermenting organism, Saccharomyces cerevisiae (T0936, a genetically-modified strain with the ability to ferment xylose, was used inside the rMBR. The rMBR was evaluated for simultaneous glucose and xylose utilization as well as in situ detoxification of furfural and hydroxylmethyl furfural (HMF. The synthetic medium was investigated, after which the pretreated wheat straw was used as a xylose-rich lignocellulosic substrate. The IPC membrane panels were successfully used as the rMBR during the batch fermentations, which lasted for up to eight days without fouling. With the rMBR, complete glucose and xylose utilization, resulting in 86% of the theoretical ethanol yield, was observed with the synthetic medium. Its application with the pretreated wheat straw resulted in complete glucose consumption and 87% xylose utilization; a final ethanol concentration of 30.3 g/L was obtained, which corresponds to 83% of the theoretical yield. Moreover, complete in situ detoxification of furfural and HMF was obtained within 36 h and 60 h, respectively, with the rMBR. The use of the rMBR is a promising technology for large-scale lignocellulosic ethanol production, since it facilitates the co-utilization of glucose and xylose; moreover, the technology would also allow the reuse of the yeast for several batches.

Co-Utilization of Glucose and Xylose for Enhanced Lignocellulosic Ethanol Production with Reverse Membrane Bioreactors

Science.gov (United States)

Ishola, Mofoluwake M.; Ylitervo, Päivi; Taherzadeh, Mohammad J.

2015-01-01

Integrated permeate channel (IPC) flat sheet membranes were examined for use as a reverse membrane bioreactor (rMBR) for lignocellulosic ethanol production. The fermenting organism, Saccharomyces cerevisiae (T0936), a genetically-modified strain with the ability to ferment xylose, was used inside the rMBR. The rMBR was evaluated for simultaneous glucose and xylose utilization as well as in situ detoxification of furfural and hydroxylmethyl furfural (HMF). The synthetic medium was investigated, after which the pretreated wheat straw was used as a xylose-rich lignocellulosic substrate. The IPC membrane panels were successfully used as the rMBR during the batch fermentations, which lasted for up to eight days without fouling. With the rMBR, complete glucose and xylose utilization, resulting in 86% of the theoretical ethanol yield, was observed with the synthetic medium. Its application with the pretreated wheat straw resulted in complete glucose consumption and 87% xylose utilization; a final ethanol concentration of 30.3 g/L was obtained, which corresponds to 83% of the theoretical yield. Moreover, complete in situ detoxification of furfural and HMF was obtained within 36 h and 60 h, respectively, with the rMBR. The use of the rMBR is a promising technology for large-scale lignocellulosic ethanol production, since it facilitates the co-utilization of glucose and xylose; moreover, the technology would also allow the reuse of the yeast for several batches. PMID:26633530

Signature pathway expression of xylose utilization in the genetically engineered industrial yeast Saccharomyces cerevisiae

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Background: The limited xylose utilizing ability of native Saccharomyces cerevisiae has been a major obstacle for efficient cellulosic ethanol production from lignocellulosic materials. Haploid laboratory strains of S. cerevisiae are commonly used for genetic engineering to enable its xylose utiliza...

Utilization of xylose as a carbon source for mixotrophic growth of Scenedesmus obliquus.

Science.gov (United States)

Yang, Suling; Liu, Guijun; Meng, Youting; Wang, Ping; Zhou, Sijing; Shang, Hongzhong

2014-11-01

Mixotrophic cultivation is one potential mode for microalgae production, and an economically acceptable and environmentally sustainable organic carbon source is essential. The potential use of xylose for culturing Scenedesmus obliquus in a mixotrophic mode and physiological features of xylose-grown S. obliquus were studied. S. obliquus had a certain xylose tolerance, and was capable of utilizing xylose for growth. At a xylose concentration of 4gL(-1), the maximal cell density was 2.2gL(-1), being 2.9-fold of that under photoautotrophic condition and arriving to the level of mixotrophic growth using 4gL(-1) glucose. No changes in cellular morphology of the cells grown with or without xylose were detected. Fluorescence emission from photosystem II (PS II) relative to photosystem I (PS I) was decreased in mixotrophic cells, implying that the PSII activity was decreased. The biomass lipid content was enhanced and carbohydrate concentration was decreased, in relation to photoautotrophic controls. Copyright © 2014 Elsevier Ltd. All rights reserved.

Conversion of xylose to ethanol under aerobic conditions by Candida tropicalis

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T. W. Jeffries

1981-01-01

Candida tropicalis converts xylose to ethanol under aerobic, but not anaerobic, conditions. Ethanol production lags behind growth and is accelerated by increased aeration. Adding xylose to active cultures stimulates ethanol production as does serial subculture in a medium containing xylose as a sole carbon source.

Evolutionary engineering strategies to enhance tolerance of xylose utilizing recombinant yeast to inhibitors derived from spruce biomass

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Koppram Rakesh

2012-05-01

Full Text Available Abstract Background One of the crucial factors for a sustainable and economical production of lignocellulosic based bioethanol is the availability of a robust fermenting microorganism with high tolerance to inhibitors generated during the pretreatment of lignocellulosic raw materials, since these inhibitors are known to severely hinder growth and fermentation. Results A long-term adaptation in repetitive batch cultures in shake flasks using a cocktail of 12 different inhibitors and a long-term chemostat adaptation using spruce hydrolysate were used as evolutionary engineering strategies to improve the inhibitor tolerance in the metabolically engineered xylose utilizing Saccharomyces cerevisiae strain, TMB3400. The yeast was evolved for a period of 429 and 97 generations in repetitive batch cultures and chemostat cultivation, respectively. During the evolutionary engineering in repetitive batch cultures the maximum specific growth rate increased from 0.18 h-1 to 0.33 h-1 and the time of lag phase was decreased from 48 h to 24 h. In the chemostat adaptation, after 97 generations, the specific conversion rates of HMF and furfural were found to be 3.5 and 4 folds higher respectively, compared to rates after three generations. Two evolved strains (RK60-5, RKU90-3 and one evolved strain (KE1-17 were isolated from evolutionary engineering in repetitive batches and chemostat cultivation, respectively. The strains displayed significantly improved growth performance over TMB3400 when cultivated in spruce hydrolysate under anaerobic conditions, the evolved strains exhibited 25 to 38% increase in specific consumption rate of sugars and 32 to 50% increased specific ethanol productivity compared to TMB3400. The evolved strains RK60-5 and RKU90-3 were unable to consume xylose under anaerobic conditions, whereas, KE1-17 was found to consume xylose at similar rates as TMB3400. Conclusion Using evolutionary engineering strategies in batch and chemostat

Bacterial xylose isomerases from the mammal gut Bacteroidetes cluster function in Saccharomyces cerevisiae for effective xylose fermentation.

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Peng, Bingyin; Huang, Shuangcheng; Liu, Tingting; Geng, Anli

2015-05-17

Xylose isomerase (XI) catalyzes the conversion of xylose to xylulose, which is the key step for anaerobic ethanolic fermentation of xylose. Very few bacterial XIs can function actively in Saccharomyces cerevisiae. Here, we illustrate a group of XIs that would function for xylose fermentation in S. cerevisiae through phylogenetic analysis, recombinant yeast strain construction, and xylose fermentation. Phylogenetic analysis of deposited XI sequences showed that XI evolutionary relationship was highly consistent with the bacterial taxonomic orders and quite a few functional XIs in S. cerevisiae were clustered with XIs from mammal gut Bacteroidetes group. An XI from Bacteroides valgutus in this cluster was actively expressed in S. cerevisiae with an activity comparable to the fungal XI from Piromyces sp. Two XI genes were isolated from the environmental metagenome and they were clustered with XIs from environmental Bacteroidetes group. These two XIs could not be expressed in yeast with activity. With the XI from B. valgutus expressed in S. cerevisiae, background yeast strains were optimized by pentose metabolizing pathway enhancement and adaptive evolution in xylose medium. Afterwards, more XIs from the mammal gut Bacteroidetes group, including those from B. vulgatus, Tannerella sp. 6_1_58FAA_CT1, Paraprevotella xylaniphila and Alistipes sp. HGB5, were individually transformed into S. cerevisiae. The known functional XI from Orpinomyces sp. ukk1, a mammal gut fungus, was used as the control. All the resulting recombinant yeast strains were able to ferment xylose. The respiration-deficient strains harboring B. vulgatus and Alistipes sp. HGB5 XI genes respectively obtained specific xylose consumption rate of 0.662 and 0.704 g xylose gcdw(-1) h(-1), and ethanol specific productivity of 0.277 and 0.283 g ethanol gcdw(-1) h(-1), much comparable to those obtained by the control strain carrying Orpinomyces sp. ukk1 XI gene. This study demonstrated that XIs clustered in the

Engineering of carbon catabolite repression in recombinant xylose fermenting Saccharomyces cerevisiae

DEFF Research Database (Denmark)

Roca, Christophe Francois Aime; Haack, Martin Brian; Olsson, Lisbeth

2004-01-01

analysed for changes in xylose consumption rate and ethanol production rate during anaerobic batch and chemostat cultivations on a mixture of 20 g l(-1) glucose and 50 g l(-1) xylose, and their characteristics were compared to the parental strain S. cerevisiae TMB3001 (XYL1, XYL2, XKS1). Improvement...... that xylose is a repressive sugar for S. cerevisiae....

Pulsed addition of HMF and furfural to batch-grown xylose-utilizing Saccharomyces cerevisiae results in different physiological responses in glucose and xylose consumption phase

Science.gov (United States)

2013-01-01

nitrogen assimilation were induced 1 hour after pulsing. Conclusions The redox and energy metabolism were found to be more severely affected after pulsing of furan aldehydes during the xylose consumption phase than during glucose consumption. Conceivably, this discrepancy resulted from the low xylose utilization rate, hence suggesting that xylose metabolism is a feasible target for metabolic engineering of more robust xylose-utilizing yeast strains. PMID:24341320

Dynamic flux balance modeling of microbial co-cultures for efficient batch fermentation of glucose and xylose mixtures.

Science.gov (United States)

Hanly, Timothy J; Henson, Michael A

2011-02-01

Sequential uptake of pentose and hexose sugars that compose lignocellulosic biomass limits the ability of pure microbial cultures to efficiently produce value-added bioproducts. In this work, we used dynamic flux balance modeling to examine the capability of mixed cultures of substrate-selective microbes to improve the utilization of glucose/xylose mixtures and to convert these mixed substrates into products. Co-culture simulations of Escherichia coli strains ALS1008 and ZSC113, engineered for glucose and xylose only uptake respectively, indicated that improvements in batch substrate consumption observed in previous experimental studies resulted primarily from an increase in ZSC113 xylose uptake relative to wild-type E. coli. The E. coli strain ZSC113 engineered for the elimination of glucose uptake was computationally co-cultured with wild-type Saccharomyces cerevisiae, which can only metabolize glucose, to determine if the co-culture was capable of enhanced ethanol production compared to pure cultures of wild-type E. coli and the S. cerevisiae strain RWB218 engineered for combined glucose and xylose uptake. Under the simplifying assumption that both microbes grow optimally under common environmental conditions, optimization of the strain inoculum and the aerobic to anaerobic switching time produced an almost twofold increase in ethanol productivity over the pure cultures. To examine the effect of reduced strain growth rates at non-optimal pH and temperature values, a break even analysis was performed to determine possible reductions in individual strain substrate uptake rates that resulted in the same predicted ethanol productivity as the best pure culture. © 2010 Wiley Periodicals, Inc.

Ethanol production by recombinant and natural xylose-utilising yeasts

Energy Technology Data Exchange (ETDEWEB)

Eliasson, Anna

2000-07-01

The xylose-fermenting capacity of recombinant Saccharomyces cerevisiae carrying XYL1 and XYL2 from Pichia stipitis, which encode xylose reductase (XR) and xylitol dehydrogenase (XDH), respectively, is poor due to high xylitol formation. Whereas, P. stipitis exhibits high ethanol yield on xylose, the tolerance towards inhibitors in the lignocellulosic hydrolysate is low. A recombinant strain possessing the advantageous characteristics of both S. cerevisiae and P. stipitis would constitute a biocatalyst capable of efficient ethanol production from lignocellulosic hydrolysate. In the work presented in this thesis, factors influencing xylose fermentation in recombinant S. cerevisiae and in the natural xylose-fermenting yeast P. stipitis have been identified and investigated. Anaerobic xylulose fermentation was compared in strains of Zygosaccharomyces and S. cerevisiae, mutants and wild-type strains to identify host strain background and genetic modifications beneficial for xylose fermentation. The greatest positive effect was found for over-expression of the gene XKS1 for the pentose phosphate pathway (PPP) enzyme xylulokinase (XK), which increased the ethanol yield by almost 85%. The Zygosaccharomyces strains tested formed large amounts of polyols, making them unsuitable as host strains. The XR/XDH/XK ratio was found to determine whether carbon accumulated in a xylitol pool or was further utilised for ethanol production in recombinant xylose-utilising S. cerevisiae. Simulations, based on a kinetic model, and anaerobic xylose cultivation experiments implied that a 1:{>=}10:{>=}4 relation was optimal in minimising xylitol formation. Ethanol formation increased with decreasing XR/XDH ratio, whereas xylitol formation decreased and XK overexpression was necessary for adequate ethanol formation. Based on the knowledge of optimal enzyme ratios, a stable, xylose-utilising strain, S. cerevisiae TMB 3001, was constructed by chromosomal integration of the XYL1 and XYL2 genes

Combined enzyme mediated fermentation of cellulose and xylose to ethanol by Schizosaccharomyces pombe, cellulase, [beta]-glucosidase, and xylose isomerase

Science.gov (United States)

Lastick, S.M.; Mohagheghi, A.; Tucker, M.P.; Grohmann, K.

1994-12-13

A process for producing ethanol from mixed sugar streams from pretreated biomass comprising xylose and cellulose using enzymes to convert these substrates to fermentable sugars; selecting and isolating a yeast Schizosaccharomyces pombe ATCC No. 2476, having the ability to ferment these sugars as they are being formed to produce ethanol; loading the substrates with the fermentation mix composed of yeast, enzymes and substrates; fermenting the loaded substrates and enzymes under anaerobic conditions at a pH range of between about 5.0 to about 6.0 and at a temperature range of between about 35 C to about 40 C until the fermentation is completed, the xylose being isomerized to xylulose, the cellulose being converted to glucose, and these sugars being concurrently converted to ethanol by yeast through means of the anaerobic fermentation; and recovering the ethanol. 2 figures.

A strict anaerobic extreme thermophilic hydrogen-producing culture enriched from digested household waste

DEFF Research Database (Denmark)

Karakashev, Dimitar Borisov; Kotay, Shireen Meher; Trably, Eric

2009-01-01

The aim of this study was to enrich, characterize and identify strict anaerobic extreme thermophilic hydrogen (H-2) producers from digested household solid wastes. A strict anaerobic extreme thermophilic H-2 producing bacterial culture was enriched from a lab-scale digester treating household...... wastes at 70 degrees C. The enriched mixed culture consisted of two rod-shaped bacterial members growing at an optimal temperature of 80 degrees C and an optimal pH 8.1. The culture was able to utilize glucose, galactose, mannose, xylose, arabinose, maltose, sucrose, pyruvate and glycerol as carbon...... sources. Growth on glucose produced acetate, H-2 and carbon dioxide. Maximal H-2 production rate on glucose was 1.1 mmol l(-1) h(-1) with a maximum H-2 yield of 1.9 mole H-2 per mole glucose. 16S ribosomal DNA clone library analyses showed that the culture members were phylogenetically affiliated...

Separate hydrolysis and co-fermentation for improved xylose utilization in integrated ethanol production from wheat meal and wheat straw

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Erdei Borbála

2012-03-01

Full Text Available Abstract Background The commercialization of second-generation bioethanol has not been realized due to several factors, including poor biomass utilization and high production cost. It is generally accepted that the most important parameters in reducing the production cost are the ethanol yield and the ethanol concentration in the fermentation broth. Agricultural residues contain large amounts of hemicellulose, and the utilization of xylose is thus a plausible way to improve the concentration and yield of ethanol during fermentation. Most naturally occurring ethanol-fermenting microorganisms do not utilize xylose, but a genetically modified yeast strain, TMB3400, has the ability to co-ferment glucose and xylose. However, the xylose uptake rate is only enhanced when the glucose concentration is low. Results Separate hydrolysis and co-fermentation of steam-pretreated wheat straw (SPWS combined with wheat-starch hydrolysate feed was performed in two separate processes. The average yield of ethanol and the xylose consumption reached 86% and 69%, respectively, when the hydrolysate of the enzymatically hydrolyzed (18.5% WIS unwashed SPWS solid fraction and wheat-starch hydrolysate were fed to the fermentor after 1 h of fermentation of the SPWS liquid fraction. In the other configuration, fermentation of the SPWS hydrolysate (7.0% WIS, resulted in an average ethanol yield of 93% from fermentation based on glucose and xylose and complete xylose consumption when wheat-starch hydrolysate was included in the feed. Increased initial cell density in the fermentation (from 5 to 20 g/L did not increase the ethanol yield, but improved and accelerated xylose consumption in both cases. Conclusions Higher ethanol yield has been achieved in co-fermentation of xylose and glucose in SPWS hydrolysate when wheat-starch hydrolysate was used as feed, then in co-fermentation of the liquid fraction of SPWS fed with the mixed hydrolysates. Integration of first-generation and

Xylose utilizing zymomonas mobilis with improved ethanol production in biomass hydrolysate medium

Science.gov (United States)

Caimi, Perry G; Hitz, William D; Stieglitz, Barry; Viitanen, Paul V

2013-07-02

Xylose-utilizing, ethanol producing strains of Zymomonas mobilis with improved performance in medium comprising biomass hydrolysate were isolated using an adaptation process. Independently isolated strains were found to have independent mutations in the same coding region. Mutation in this coding may be engineered to confer the improved phenotype.

Highly enriched Betaproteobacteria growing anaerobically with p-xylene and nitrate

DEFF Research Database (Denmark)

Rotaru, Amelia-Elena; Probian, Christina; Wilkes, Heinz

2010-01-01

The identity of the microorganisms capable of anaerobic p-xylene degradation under denitrifying conditions is hitherto unknown. Here, we report highly enriched cultures of freshwater denitrifying bacteria that grow anaerobically with p-xylene as the sole organic carbon source and electron donor. ...

Heterologous expression of Spathaspora passalidarum xylose reductase and xylitol dehydrogenase genes improved xylose fermentation ability of Aureobasidium pullulans.

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Guo, Jian; Huang, Siyao; Chen, Yefu; Guo, Xuewu; Xiao, Dongguang

2018-04-30

Aureobasidium pullulans is a yeast-like fungus that can ferment xylose to generate high-value-added products, such as pullulan, heavy oil, and melanin. The combinatorial expression of two xylose reductase (XR) genes and two xylitol dehydrogenase (XDH) genes from Spathaspora passalidarum and the heterologous expression of the Piromyces sp. xylose isomerase (XI) gene were induced in A. pullulans to increase the consumption capability of A. pullulans on xylose. The overexpression of XYL1.2 (encoding XR) and XYL2.2 (encoding XDH) was the most beneficial for xylose utilization, resulting in a 17.76% increase in consumed xylose compared with the parent strain, whereas the introduction of the Piromyces sp. XI pathway failed to enhance xylose utilization efficiency. Mutants with superior xylose fermentation performance exhibited increased intracellular reducing equivalents. The fermentation performance of all recombinant strains was not affected when glucose or sucrose was utilized as the carbon source. The strain with overexpression of XYL1.2 and XYL2.2 exhibited excellent fermentation performance with mimicked hydrolysate, and pullulan production increased by 97.72% compared with that of the parent strain. The present work indicates that the P4 mutant (using the XR/XDH pathway) with overexpressed XYL1.2 and XYL2.2 exhibited the best xylose fermentation performance. The P4 strain showed the highest intracellular reducing equivalents and XR and XDH activity, with consequently improved pullulan productivity and reduced melanin production. This valuable development in aerobic fermentation by the P4 strain may provide guidance for the biotransformation of xylose to high-value products by A. pullulans through genetic approach.

Enhanced isoprenoid production from xylose by engineered Saccharomyces cerevisiae.

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Kwak, Suryang; Kim, Soo Rin; Xu, Haiqing; Zhang, Guo-Chang; Lane, Stephan; Kim, Heejin; Jin, Yong-Su

2017-11-01

Saccharomyces cerevisiae has limited capabilities for producing fuels and chemicals derived from acetyl-CoA, such as isoprenoids, due to a rigid flux partition toward ethanol during glucose metabolism. Despite numerous efforts, xylose fermentation by engineered yeast harboring heterologous xylose metabolic pathways was not as efficient as glucose fermentation for producing ethanol. Therefore, we hypothesized that xylose metabolism by engineered yeast might be a better fit for producing non-ethanol metabolites. We indeed found that engineered S. cerevisiae on xylose showed higher expression levels of the enzymes involved in ethanol assimilation and cytosolic acetyl-CoA synthesis than on glucose. When genetic perturbations necessary for overproducing squalene and amorphadiene were introduced into engineered S. cerevisiae capable of fermenting xylose, we observed higher titers and yields of isoprenoids under xylose than glucose conditions. Specifically, co-overexpression of a truncated HMG1 (tHMG1) and ERG10 led to substantially higher squalene accumulation under xylose than glucose conditions. In contrast to glucose utilization producing massive amounts of ethanol regardless of aeration, xylose utilization allowed much less amounts of ethanol accumulation, indicating ethanol is simultaneously re-assimilated with xylose consumption and utilized for the biosynthesis of cytosolic acetyl-CoA. In addition, xylose utilization by engineered yeast with overexpression of tHMG1, ERG10, and ADS coding for amorphadiene synthase, and the down-regulation of ERG9 resulted in enhanced amorphadiene production as compared to glucose utilization. These results suggest that the problem of the rigid flux partition toward ethanol production in yeast during the production of isoprenoids and other acetyl-CoA derived chemicals can be bypassed by using xylose instead of glucose as a carbon source. Biotechnol. Bioeng. 2017;114: 2581-2591. © 2017 Wiley Periodicals, Inc. © 2017 Wiley

Analysis of metabolisms and transports of xylitol using xylose- and xylitol-assimilating Saccharomyces cerevisiae.

Science.gov (United States)

Tani, Tatsunori; Taguchi, Hisataka; Akamatsu, Takashi

2017-05-01

To clarify the relationship between NAD(P) + /NAD(P)H redox balances and the metabolisms of xylose or xylitol as carbon sources, we analyzed aerobic and anaerobic batch cultures of recombinant Saccharomyces cerevisiae in a complex medium containing 20 g/L xylose or 20 g/L xylitol at pH 5.0 and 30°C. The TDH3p-GAL2 or gal80Δ strain completely consumed the xylose within 24 h and aerobically consumed 92-100% of the xylitol within 96 h, but anaerobically consumed only 20% of the xylitol within 96 h. Cells of both strains grew well in aerobic culture. The addition of acetaldehyde (an effective oxidizer of NADH) increased the xylitol consumption by the anaerobically cultured strain. These results indicate that in anaerobic culture, NAD + generated in the NAD(P)H-dependent xylose reductase reaction was likely needed in the NAD + -dependent xylitol dehydrogenase reaction, whereas in aerobic culture, the NAD + generated by oxidation of NADH in the mitochondria is required in the xylitol dehydrogenase reaction. The role of Gal2 and Fps1 in importing xylitol into the cytosol and exporting it from the cells was analyzed by examining the xylitol consumption in aerobic culture and the export of xylitol metabolized from xylose in anaerobic culture, respectively. The xylitol consumptions of gal80Δ gal2Δ and gal80Δ gal2Δ fps1Δ strains were reduced by 81% and 88% respectively, relative to the gal80Δ strain. The maximum xylitol concentration accumulated by the gal80Δ, gal80Δ gal2Δ, and gal80Δ gal2Δ fps1Δ strains was 7.25 g/L, 5.30 g/L, and 4.27 g/L respectively, indicating that Gal2 and Fps1 transport xylitol both inward and outward. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Comparison of heterologous xylose transporters in recombinant Saccharomyces cerevisiae

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Hahn-Hägerdal Bärbel

2010-03-01

Full Text Available Abstract Background Baker's yeast (Saccharomyces cerevisiae has been engineered for xylose utilization to enable production of fuel ethanol from lignocellulose raw material. One unresolved challenge is that S. cerevisiae lacks a dedicated transport system for pentose sugars, which means that xylose is transported by non-specific Hxt transporters with comparatively low transport rate and affinity for xylose. Results In this study, we compared three heterologous xylose transporters that have recently been shown to improve xylose uptake under different experimental conditions. The transporters Gxf1, Sut1 and At5g59250 from Candida intermedia, Pichia stipitis and Arabidopsis thaliana, respectively, were expressed in isogenic strains of S. cerevisiae and the transport kinetics and utilization of xylose was evaluated. Expression of the Gxf1 and Sut1 transporters led to significantly increased affinity and transport rates of xylose. In batch cultivation at 4 g/L xylose concentration, improved transport kinetics led to a corresponding increase in xylose utilization, whereas no correlation could be demonstrated at xylose concentrations greater than 15 g/L. The relative contribution of native sugar transporters to the overall xylose transport capacity was also estimated during growth on glucose and xylose. Conclusions Kinetic characterization and aerobic batch cultivation of strains expressing the Gxf1, Sut1 and At5g59250 transporters showed a direct relationship between transport kinetics and xylose growth. The Gxf1 transporter had the highest transport capacity and the highest xylose growth rate, followed by the Sut1 transporter. The range in which transport controlled the growth rate was determined to between 0 and 15 g/L xylose. The role of catabolite repression in regulation of native transporters was also confirmed by the observation that xylose transport by native S. cerevisiae transporters increased significantly during cultivation in xylose and

Formation of xylitol and xylitol-5-phosphate and its impact on growth of d-xylose-utilizing Corynebacterium glutamicum strains.

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Radek, Andreas; Müller, Moritz-Fabian; Gätgens, Jochem; Eggeling, Lothar; Krumbach, Karin; Marienhagen, Jan; Noack, Stephan

2016-08-10

Wild-type Corynebacterium glutamicum has no endogenous metabolic activity for utilizing the lignocellulosic pentose d-xylose for cell growth. Therefore, two different engineering approaches have been pursued resulting in platform strains harbouring a functional version of either the Isomerase (ISO) or the Weimberg (WMB) pathway for d-xylose assimilation. In a previous study we found for C. glutamicum WMB by-product formation of xylitol during growth on d-xylose and speculated that the observed lower growth rates are due to the growth inhibiting effect of this compound. Based on a detailed phenotyping of the ISO, WMB and the wild-type strain of C. glutamicum, we here show that this organism has a natural capability to synthesize xylitol from d-xylose under aerobic cultivation conditions. We furthermore observed the intracellular accumulation of xylitol-5-phosphate as a result of the intracellular phosphorylation of xylitol, which was particularly pronounced in the C. glutamicum ISO strain. Interestingly, low amounts of supplemented xylitol strongly inhibit growth of this strain on d-xylose, d-glucose and d-arabitol. These findings demonstrate that xylitol is a suitable substrate of the endogenous xylulokinase (XK, encoded by xylB) and its overexpression in the ISO strain leads to a significant phosphorylation of xylitol in C. glutamicum. Therefore, in order to circumvent cytotoxicity by xylitol-5-phosphate, the WMB pathway represents an interesting alternative route for engineering C. glutamicum towards efficient d-xylose utilization. Copyright © 2016 Elsevier B.V. All rights reserved.

Comparison of the xylose reductase-xylitol dehydrogenase and the xylose isomerase pathways for xylose fermentation by recombinant Saccharomyces cerevisiae

Directory of Open Access Journals (Sweden)

Hahn-Hägerdal Bärbel

2007-02-01

Full Text Available Abstract Background Two heterologous pathways have been used to construct recombinant xylose-fermenting Saccharomyces cerevisiae strains: i the xylose reductase (XR and xylitol dehydrogenase (XDH pathway and ii the xylose isomerase (XI pathway. In the present study, the Pichia stipitis XR-XDH pathway and the Piromyces XI pathway were compared in an isogenic strain background, using a laboratory host strain with genetic modifications known to improve xylose fermentation (overexpressed xylulokinase, overexpressed non-oxidative pentose phosphate pathway and deletion of the aldose reductase gene GRE3. The two isogenic strains and the industrial xylose-fermenting strain TMB 3400 were studied regarding their xylose fermentation capacity in defined mineral medium and in undetoxified lignocellulosic hydrolysate. Results In defined mineral medium, the xylose consumption rate, the specific ethanol productivity, and the final ethanol concentration were significantly higher in the XR- and XDH-carrying strain, whereas the highest ethanol yield was achieved with the strain carrying XI. While the laboratory strains only fermented a minor fraction of glucose in the undetoxified lignocellulose hydrolysate, the industrial strain TMB 3400 fermented nearly all the sugar available. Xylitol was formed by the XR-XDH-carrying strains only in mineral medium, whereas in lignocellulose hydrolysate no xylitol formation was detected. Conclusion Despite by-product formation, the XR-XDH xylose utilization pathway resulted in faster ethanol production than using the best presently reported XI pathway in the strain background investigated. The need for robust industrial yeast strains for fermentation of undetoxified spruce hydrolysates was also confirmed.

Overview of Catalytic Properties of Fungal Xylose Reductases and Molecular Engineering Approaches for Improved Xylose Utilisation in Yeast

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Sk Amir Hossain

2018-03-01

Full Text Available Background and Objective: Xylose reductases belong to the aldo-keto reductase family of enzymes, which catalyse the conversion of xylose to xylitol. Yeast xylose reductases have been intensively studied in the last two decades due to their significance in biotechnological production of ethanol and xylitol from xylose. Due to its GRAS status and pronounced tolerance to harsh conditions, Saccharomyces cerevisiae is the ideal organism for industrial production of both xylitol and ethanol. However, Saccharomyces cerevisiae is unable to use xylose as the sole carbon source due to the lack of xylose specific transporters and insufficient activity of metabolic pathways for xylose utilisation. The aim of this paper is to give an overview of attempts in increasing biotechnological potential of xylose reductases and to highlight the prospective of this application. Results and Conclusion: In order to create strains with improved xylose utilization, different approaches were attempted including simultaneous overexpression of xylitol dehydrogenase, xylose reductase and pentose phosphate pathway enzymes, heterologous expression of putative xylose transporters or heterologous expression of genes coding for enzymes included in the xylose metabolism, respectively. Furthermore, number of attempts to genetically modify different xylose reductases is increasing. This review presents current knowledge about yeast xylose reductases and the different approaches applied in order to improve xylose metabolism in yeast.Conflict of interest: The authors declare no conflict of interest.

xylA and xylB overexpression as a successful strategy for improving xylose utilization and poly-3-hydroxybutyrate production in Burkholderia sacchari.

Science.gov (United States)

Guamán, Linda P; Oliveira-Filho, Edmar R; Barba-Ostria, Carlos; Gomez, José G C; Taciro, Marilda K; da Silva, Luiziana Ferreira

2018-03-01

Despite the versatility and many advantages of polyhydroxyalkanoates as petroleum-based plastic substitutes, their higher production cost compared to petroleum-based polymers has historically limited their large-scale production. One appealing approach to reducing production costs is to employ less expensive, renewable feedstocks. Xylose, for example is an abundant and inexpensive carbon source derived from hemicellulosic residues abundant in agro-industrial waste (sugarcane bagasse hemicellulosic hydrolysates). In this work, the production of poly-3-hydroxybutyrate P(3HB) from xylose was studied to develop technologies for conversion of agro-industrial waste into high-value chemicals and biopolymers. Specifically, this work elucidates the organization of the xylose assimilation operon of Burkholderia sacchari, a non-model bacterium with high capacity for P(3HB) accumulation. Overexpression of endogenous xylose isomerase and xylulokinase genes was successfully assessed, improving both specific growth rate and P(3HB) production. Compared to control strain (harboring pBBR1MCS-2), xylose utilization in the engineered strain was substantially improved with 25% increase in specific growth rate, 34% increase in P(3HB) production, and the highest P(3HB) yield from xylose reported to date for B. sacchari (Y P3HB/Xil  = 0.35 g/g). This study highlights that xylA and xylB overexpression is an effective strategy to improve xylose utilization and P(3HB) production in B. sacchari.

Pentose utilization in yeasts: Physiology and biochemistry

Energy Technology Data Exchange (ETDEWEB)

Jeppson, H.

1996-04-01

The fermentive performance of bacteria, yeasts, and filamentous fungi was investigated in a pentose (xylose)-rich lignocellulosic hydrolyzate. The filamentous fungus Fusarium oxysporum and the xylose-fermenting yeast Pichia stipitis were found to be very sensitive to the inhibiting hydrolyzate. Recombinant xylose-utilizing Saccharomyces cerevisiae showed very poor ethanol formation from xylose; xylitol being the major product formed. The highest ethanol yields were obtained with recombinant Escherichia coli KO11, however, for maximal ethanol yield detoxification of the hydrolyzate was required. The influence of oxygen on the regulation of carbohydrate metabolism in the xylose-fermenting yeast P. stipitis CBS 6054 was investigated. A low and well-controlled level of oxygenation has been found to be required for efficient ethanol formation from xylose by the xylose-fermenting yeasts. The requirement of oxygen is frequently ascribed to the apparent redox imbalance which develops under anaerobic conditions due to the difference in co-factor utilization of the two first enzymes in the xylose metabolism, further reflected in xylitol excretion. However, a low and well controlled level of oxygenation for maximal ethanol production from glucose was also demonstrated, suggesting that the oxygen requirement is not only due to the dual co-factor utilization, but also serves other purposes. Cyanide-insensitive and salicyl hydroxamic acid-sensitive respiration (CIR) was found in P. stipitis. CIR is suggested to act as a redox sink preventing xylitol formation in P. stipitis under oxygen-limited xylose fermentations. Xylitol metabolism by P. stipitis CBS 6054 was strictly respiratory and ethanol was not formed under any conditions. The absence of ethanol formation was not due to a lack of fermentative enzymes, since the addition of glucose to xylitol-pregrown cells resulted in ethanol formation. 277 refs, 5 figs, 7 tabs

Thermophilic anaerobic acetate-utilizing methanogens and their metabolism

DEFF Research Database (Denmark)

Mladenovska, Zuzana

Six strains of thermophilic anaerobic acetate-utilizing methanogens were isolated from different full-scale thermophilic biogas plants in China and Denmark. The strain isolated from the Chinese biogas plant was designated KN-6P and the isolates from the Danish full-scale biogas plants were......, utilizing the substrates acetate, methanol and methylamines but not hydrogen/carbon dioxide. Strain Methanosarcina sp. SO-2P was able to grow mixotrophically on methanol and hydrogen/carbon dioxide with methane formation from hydrogen and carbon dioxide occurring after methanol depletion. All six...... designated HG-1P, LVG-4P R1-1P, SO-2P and V-1P. The isolates were characterized morphologically and physiologically, and their immunological and phylogenetic relatedness to already known isolated strains were established. All isolated strains were identified as organisms belonging to genus Methanosarcina...

Selection of yeast Saccharomyces cerevisiae promoters available for xylose cultivation and fermentation.

Science.gov (United States)

Nambu-Nishida, Yumiko; Sakihama, Yuri; Ishii, Jun; Hasunuma, Tomohisa; Kondo, Akihiko

2018-01-01

To efficiently utilize xylose, a major sugar component of hemicelluloses, in Saccharomyces cerevisiae requires the proper expression of varied exogenous and endogenous genes. To expand the repertoire of promoters in engineered xylose-utilizing yeast strains, we selected promoters in S. cerevisiae during cultivation and fermentation using xylose as a carbon source. To select candidate promoters that function in the presence of xylose, we performed comprehensive gene expression analyses using xylose-utilizing yeast strains both during xylose and glucose fermentation. Based on microarray data, we chose 29 genes that showed strong, moderate, and weak expression in xylose rather than glucose fermentation. The activities of these promoters in a xylose-utilizing yeast strain were measured by lacZ reporter gene assays over time during aerobic cultivation and microaerobic fermentation, both in xylose and glucose media. In xylose media, P TDH3 , P FBA1 , and P TDH1 were favorable for high expression, and P SED1 , P HXT7 , P PDC1 , P TEF1 , P TPI1 , and P PGK1 were acceptable for medium-high expression in aerobic cultivation, and moderate expression in microaerobic fermentation. P TEF2 allowed moderate expression in aerobic culture and weak expression in microaerobic fermentation, although it showed medium-high expression in glucose media. P ZWF1 and P SOL4 allowed moderate expression in aerobic cultivation, while showing weak but clear expression in microaerobic fermentation. P ALD3 and P TKL2 showed moderate promoter activity in aerobic cultivation, but showed almost no activity in microaerobic fermentation. The knowledge of promoter activities in xylose cultivation obtained in this study will permit the control of gene expression in engineered xylose-utilizing yeast strains that are used for hemicellulose fermentation. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Alcohol Fermentation and Biomass formation from xylose, glucose ...

African Journals Online (AJOL)

Cerevisiae (LB-7) was the slowest in growth and utilization of xylose into biomass (economic conversion coefficient of 0.03), while K3 showed fastest utilization of xylose (coefficient 0.76). For the production of ethanol, the fastest growth and assimilation of glucose was recorded by Pa. tannophilus (P1) (coefficient 0.56) ...

The utility of anaerobic blood culture in detecting facultative anaerobic bacteremia in children.

Science.gov (United States)

Shoji, Kensuke; Komuro, Hisako; Watanabe, Yasushi; Miyairi, Isao

2013-08-01

Routine anaerobic blood culture is not recommended in children because obligate anaerobic bacteremia is rare in the pediatric population. However, a number of facultative anaerobic bacteria can cause community and hospital acquired infections in children and the utility of anaerobic blood culture for detection of these organisms is still unclear. We conducted a retrospective analysis of all blood culture samples (n = 24,356) at a children's hospital in Japan from October 2009 to June 2012. Among the samples that had paired aerobic and anaerobic blood cultures, 717 samples were considered clinically significant with 418 (58%) organisms detected from both aerobic and anaerobic cultures, 167 (23%) detected only from aerobic culture and 132 (18%) detected only from anaerobic culture. While most facultative anaerobes were detectable by aerobic culture, over 25% of Enterobacteriaceae and 15% of Staphylococcus sp. were detected from anaerobic cultures bottles only, suggesting its potential role in selected settings. Copyright © 2013 Elsevier Inc. All rights reserved.

Comparative genomics of xylose-fermenting fungi for enhanced biofuel production

Energy Technology Data Exchange (ETDEWEB)

Wohlbach, Dana J.; Kuo, Alan; Sato, Trey K.; Potts, Katlyn M.; Salamov, Asaf A.; LaButti, Kurt M.; Sun, Hui; Clum, Alicia; Pangilinan, Jasmyn L.; Lindquist, Erika A.; Lucas, Susan; Lapidus, Alla; Jin, Mingjie; Gunawan, Christa; Balan, Venkatesh; Dale, Bruce E.; Jeffries, Thomas W.; Zinkel, Robert; Barry, Kerrie W.; Grigoriev, Igor V.; Gasch, Audrey P.

2011-02-24

Cellulosic biomass is an abundant and underused substrate for biofuel production. The inability of many microbes to metabolize the pentose sugars abundant within hemicellulose creates specific challenges for microbial biofuel production from cellulosic material. Although engineered strains of Saccharomyces cerevisiae can use the pentose xylose, the fermentative capacity pales in comparison with glucose, limiting the economic feasibility of industrial fermentations. To better understand xylose utilization for subsequent microbial engineering, we sequenced the genomes of two xylose-fermenting, beetle-associated fungi, Spathaspora passalidarum and Candida tenuis. To identify genes involved in xylose metabolism, we applied a comparative genomic approach across 14 Ascomycete genomes, mapping phenotypes and genotypes onto the fungal phylogeny, and measured genomic expression across five Hemiascomycete species with different xylose-consumption phenotypes. This approach implicated many genes and processes involved in xylose assimilation. Several of these genes significantly improved xylose utilization when engineered into S. cerevisiae, demonstrating the power of comparative methods in rapidly identifying genes for biomass conversion while reflecting on fungal ecology.

Directed evolution of xylose isomerase for improved xylose catabolism and fermentation in the yeast Saccharomyces cerevisiae.

Science.gov (United States)

Lee, Sun-Mi; Jellison, Taylor; Alper, Hal S

2012-08-01

The heterologous expression of a highly functional xylose isomerase pathway in Saccharomyces cerevisiae would have significant advantages for ethanol yield, since the pathway bypasses cofactor requirements found in the traditionally used oxidoreductase pathways. However, nearly all reported xylose isomerase-based pathways in S. cerevisiae suffer from poor ethanol productivity, low xylose consumption rates, and poor cell growth compared with an oxidoreductase pathway and, additionally, often require adaptive strain evolution. Here, we report on the directed evolution of the Piromyces sp. xylose isomerase (encoded by xylA) for use in yeast. After three rounds of mutagenesis and growth-based screening, we isolated a variant containing six mutations (E15D, E114G, E129D, T142S, A177T, and V433I) that exhibited a 77% increase in enzymatic activity. When expressed in a minimally engineered yeast host containing a gre3 knockout and tal1 and XKS1 overexpression, the strain expressing this mutant enzyme improved its aerobic growth rate by 61-fold and both ethanol production and xylose consumption rates by nearly 8-fold. Moreover, the mutant enzyme enabled ethanol production by these yeasts under oxygen-limited fermentation conditions, unlike the wild-type enzyme. Under microaerobic conditions, the ethanol production rates of the strain expressing the mutant xylose isomerase were considerably higher than previously reported values for yeast harboring a xylose isomerase pathway and were also comparable to those of the strains harboring an oxidoreductase pathway. Consequently, this study shows the potential to evolve a xylose isomerase pathway for more efficient xylose utilization.

Dynamic metabolomics differentiates between carbon and energy starvation in recombinant Saccharomyces cerevisiae fermenting xylose

Directory of Open Access Journals (Sweden)

Bergdahl Basti

2012-05-01

Full Text Available Abstract Background The concerted effects of changes in gene expression due to changes in the environment are ultimately reflected in the metabolome. Dynamics of metabolite concentrations under a certain condition can therefore give a description of the cellular state with a high degree of functional information. We used this potential to evaluate the metabolic status of two recombinant strains of Saccharomyces cerevisiae during anaerobic batch fermentation of a glucose/xylose mixture. Two isogenic strains were studied, differing only in the pathways used for xylose assimilation: the oxidoreductive pathway with xylose reductase (XR and xylitol dehydrogenase (XDH or the isomerization pathway with xylose isomerase (XI. The isogenic relationship between the two strains ascertains that the observed responses are a result of the particular xylose pathway and not due to unknown changes in regulatory systems. An increased understanding of the physiological state of these strains is important for further development of efficient pentose-utilizing strains for bioethanol production. Results Using LC-MS/MS we determined the dynamics in the concentrations of intracellular metabolites in central carbon metabolism, nine amino acids, the purine nucleotides and redox cofactors. The general response to the transition from glucose to xylose was increased concentrations of amino acids and TCA-cycle intermediates, and decreased concentrations of sugar phosphates and redox cofactors. The two strains investigated had significantly different uptake rates of xylose which led to an enhanced response in the XI-strain. Despite the difference in xylose uptake rate, the adenylate energy charge remained high and stable around 0.8 in both strains. In contrast to the adenylate pool, large changes were observed in the guanylate pool. Conclusions The low uptake of xylose by the XI-strain led to several distinguished responses: depletion of key metabolites in glycolysis and NADPH

Engineering genome-reduced Bacillus subtilis for acetoin production from xylose.

Science.gov (United States)

Yan, Panpan; Wu, Yuanqing; Yang, Li; Wang, Zhiwen; Chen, Tao

2018-02-01

To investigate the capacity of a genome-reduced Bacillus subtilis strain as chassis cell for acetoin production from xylose. To endow the genome-reduced Bacillus subtilis strain BSK814 with the ability to utilize xylose, we inserted a native xyl operon into its genome and deleted the araR gene. The resulting strain BSK814A2 produced 2.94 g acetoin/l from 10 g xylose/l, which was 39% higher than control strain BSK19A2. The deletion of the bdhA and acoA genes further improved xylose utilization efficiency and increased acetoin production to 3.71 g/l in BSK814A4. Finally, BSK814A4 produced up to 23.3 g acetoin/l from 50 g xylose/l, with a yield of 0.46 g/g xylose. Both the titer and yield were 39% higher than those of control strain BSK19A4. As a chassis cell, genome-reduced B. subtilis showed significantly improved capacity for the production of the overflow product acetoin from xylose compared with wild-type strain.

Impact of xylose and mannose on central metabolism of yeast Saccharomyces cerevisiae

Energy Technology Data Exchange (ETDEWEB)

Pitkaenen, J.P.

2005-07-01

In this study, understanding of the central metabolism was improved by quantification of metabolite concentrations, enzyme activities, protein abundances, and gene transcript concentrations. Intracellular fluxes were estimated by applying stoichiometric models of metabolism. The methods were applied in the study of yeast Saccharomyces cerevisiae in two separate projects. A xylose project aimed at improved utilization of D- xylose as a substrate for, e.g., producing biomaterial- based fuel ethanol. A mannose project studied the production of GDP-mannose from D-mannose in a strain lacking the gene for phosphomannose isomerase (PMI40 deletion). Hexose, D-glucose is the only sugar more abundant than pentose D-xylose. D-xylose is common in hardwoods (e.g. birch) and crop residues (ca. 25% of dry weight). However, S. cerevisiae is unable to utilize D- xylose without a recombinant pathway where D-xylose is converted to Dxylulose. In this study D-xylose was converted in two steps via xylitol: by D-xylose reductase and xylitol dehydrogenase encoded by XYL1 and XYL2 from Pichia stipitis, respectively. Additionally, endogenous xylulokinase (XKS1) was overexpressed in order to increase the consumption of D-xylose by enhancing the phosphorylation of D-xylulose. Despite of the functional recombinant pathway the utilization rates of D xylose still remained low. This study proposes a set of limitations that are responsible for the low utilization rates of D-xylose under microaerobic conditions. Cells compensated for the cofactor imbalance, caused by the conversion of D-xylose to D- xylulose, by increasing the flux through the oxidative pentose phosphate pathway and by shuttling NADH redox potential to mitochondrion to be oxidized in oxidative phosphorylation. However, mitochondrial NADH inhibits citrate synthase in citric acid cycle, and consequently lower flux through citric acid cycle limits oxidative phosphorylation. Further, limitations in the uptake of D- xylose, in the

Saccharomyces cerevisiae engineered for xylose metabolism exhibits a respiratory response

Science.gov (United States)

Yong-Su Jin; Jose M. Laplaza; Thomas W. Jeffries

2004-01-01

Native strains of Saccharomyces cerevisiae do not assimilate xylose. S. cerevisiae engineered for D-xylose utilization through the heterologous expression of genes for aldose reductase ( XYL1), xylitol dehydrogenase (XYL2), and D-xylulokinase ( XYL3 or XKS1) produce only limited amounts of ethanol in xylose medium. In recombinant S. cerevisiae expressing XYL1, XYL2,...

Bioprospecting and evolving alternative xylose and arabinose pathway enzymes for use in Saccharomyces cerevisiae.

Science.gov (United States)

Lee, Sun-Mi; Jellison, Taylor; Alper, Hal S

2016-03-01

Bioprospecting is an effective way to find novel enzymes from strains with desirable phenotypes. Such bioprospecting has enabled organisms such as Saccharomyces cerevisiae to utilize nonnative pentose sugars. Yet, the efficiency of this pentose catabolism (especially for the case of arabinose) remains suboptimal. Thus, further pathway optimization or identification of novel, optimal pathways is needed. Previously, we identified a novel set of xylan catabolic pathway enzymes from a superior pentose-utilizing strain of Ustilago bevomyces. These enzymes were used to successfully engineer a xylan-utilizing S. cerevisiae through a blended approach of bioprospecting and evolutionary engineering. Here, we expanded this approach to xylose and arabinose catabolic pathway engineering and demonstrated that bioprospected xylose and arabinose catabolic pathways from U. bevomyces offer alternative choices for enabling efficient pentose catabolism in S. cerevisiae. By introducing a novel set of xylose catabolic genes from U. bevomyces, growth rates were improved up to 85 % over a set of traditional Scheffersomyces stipitis pathway genes. In addition, we suggested an alternative arabinose catabolic pathway which, after directed evolution and pathway engineering, enabled S. cerevisiae to grow on arabinose as a sole carbon source in minimal medium with growth rates upwards of 0.05 h(-1). This pathway represents the most efficient growth of yeast on pure arabinose minimal medium. These pathways provide great starting points for further strain development and demonstrate the utility of bioprospecting from U. bevomyces.

New techniques for growing anaerobic bacteria: experiments with Clostridium butyricum and Clostridium acetobutylicum

International Nuclear Information System (INIS)

Adler, H.I.; Crow, W.D.; Hadden, C.T.; Hall, J.; Machanoff, R.

1983-01-01

Stable membrane fragments derived from Escherichia coli produce and maintain strict anaerobic conditions when added to liquid or solid bacteriological media. Techniques for growing Clostridium butyricum and Clostridium acetobutylicum in membrane-containing media are described. Liquid cultures initiated by very small inocula can be grown in direct contact with air. In solid media, colonies develop rapidly from individual cells even without incubation in anaerobic jars or similar devices. Observations on growth rates, spontaneous mutations, radiation, and oxygen sensitivity of anaerobic bacteria have been made using these new techniques

[Isolation and identification of seven thermophilic and anaerobic bacteria from hot springs in Tengchong Rehai].

Science.gov (United States)

Lu, Yueqing; Chen, Bo; Liu, Xiaoli; Ji, Xiuling; Wei, Yunlin; Lin, Lianbing

2009-09-01

In order to study the taxonomic characteristic and physiological, biochemical properties of anaerobic bacteria from hot springs in Tengchong Rehai, Yunnan Province, China. Using Hungate anaerobic technique We isolated seven strains from hot springs in Tengchong Rehai, Yunnan province, and analyzed their 16S rRNA gene sequences. The seven isolates were rod-shaped, Gram-negative, obligate anaerobe, and spores formation was not observed. All strains could grow well at 70 degrees C. Growth of strain RH0802 occurred between 60 and 80 degrees C, optimally around 70 degrees C. The pH range for its growth was between 5.5 and 8.5, with an optimum around 7.0. Strain RH0802 grew on a wide range of carbon sources, including glucose, starch, mannitol, mannose, ribose, maltose, cellobiose, xylose, fructose, galactose, xylan and glycerol, but it could not utilize sucrose or pyruvate. 16S rRNA gene phylogenetic analysis showed that the maximum similarity between the five strains and the strains of genus Caldanaerobacter was up to 98%, except RH0804 and RH0806, which reached to 96% and 93%, respectively. The two isolates were presumed to be potential novel species. The GenBank accession numbers of RH0802 to RH0808 were FJ748766, FJ748762, FJ748761, FJ748763, FJ748765, FJ748764 and FJ748767. The results showed that the seven thermophilic anaerobes belonged to the genus Caldanaerobacter.

Transposon mutagenesis to improve the growth of recombinant Saccharomyces cerevisiae on D-xylose

Science.gov (United States)

Haiying Ni; Jose M. Laplaza; Thomas W. Jeffries

2007-01-01

Saccharomyces cerevisiae L2612 transformed with genes for xylose reductase and xylitol dehydrogenase (XYL1 and XYL2) grows well on glucose but very poorly on D-xylose. When a gene for D-xylulokinase (XYL3 or XKS1) is overexpressed, growth on glucose is unaffected, but growth on xylose is blocked. Spontaneous or chemically induced mutants of this engineered yeast that...

Production of xylitol by a Coniochaeta ligniaria strain tolerant of inhibitors and defective in growth on xylose.

Science.gov (United States)

Nichols, Nancy N; Saha, Badal C

2016-05-01

In conversion of biomass to fuels or chemicals, inhibitory compounds arising from physical-chemical pretreatment of the feedstock can interfere with fermentation of the sugars to product. Fungal strain Coniochaeta ligniaria NRRL30616 metabolizes the furan aldehydes furfural and 5-hydroxymethylfurfural, as well as a number of aromatic and aliphatic acids and aldehydes. Use of NRRL30616 to condition biomass sugars by metabolizing the inhibitors improves their fermentability. Wild-type C. ligniaria has the ability to grow on xylose as sole source of carbon and energy, with no accumulation of xylitol. Mutants of C. ligniaria unable to grow on xylose were constructed. Xylose reductase and xylitol dehydrogenase activities were reduced by approximately two thirds in mutant C8100. The mutant retained ability to metabolize inhibitors in biomass hydrolysates. Although C. ligniaria C8100 did not grow on xylose, the strain converted a portion of xylose to xylitol, producing 0.59 g xylitol/g xylose in rich medium and 0.48 g xylitol/g xylose in corn stover dilute acid hydrolysate. 2016 American Institute of Chemical Engineers Biotechnol. Prog., 2016 © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:606-612, 2016. © 2016 American Institute of Chemical Engineers.

Ethanol production from xylose in engineered Saccharomyces cerevisiae strains. Current state and perspectives

Energy Technology Data Exchange (ETDEWEB)

Matsushika, Akinori; Inoue, Hiroyuki; Sawayama, Shigeki [National Inst. of Advanced Industrial Science and Technology (AIST), Hiroshima (JP). Biomass Technology Research Center (BTRC); Kodaki, Tsutomu [Kyoto Univ. (Japan). Inst. of Advanced Energy

2009-08-15

Bioethanol production from xylose is important for utilization of lignocellulosic biomass as raw materials. The research on yeast conversion of xylose to ethanol has been intensively studied especially for genetically engineered Saccharomyces cerevisiae during the last 20 years. S. cerevisiae, which is a very safe microorganism that plays a traditional and major role in industrial bioethanol production, has several advantages due to its high ethanol productivity, as well as its high ethanol and inhibitor tolerance. However, this yeast cannot ferment xylose, which is the dominant pentose sugar in hydrolysates of lignocellulosic biomass. A number of different strategies have been applied to engineer yeasts capable of efficiently producing ethanol from xylose, including the introduction of initial xylose metabolism and xylose transport, changing the intracellular redox balance, and overexpression of xylulokinase and pentose phosphate pathways. In this review, recent progress with regard to these studies is discussed, focusing particularly on xylose-fermenting strains of S. cerevisiae. Recent studies using several promising approaches such as host strain selection and adaptation to obtain further improved xylose-utilizing S. cerevisiae are also addressed. (orig.)

Microaerobic conversion of xylose to ethanol in recombinant Saccharomyces cerevisiae SX6(MUT) expressing cofactor-balanced xylose metabolic enzymes and deficient in ALD6.

Science.gov (United States)

Jo, Sung-Eun; Seong, Yeong-Je; Lee, Hyun-Soo; Lee, Soo Min; Kim, Soo-Jung; Park, Kyungmoon; Park, Yong-Cheol

2016-06-10

Xylose is a major monosugar in cellulosic biomass and should be utilized for cost-effective ethanol production. In this study, xylose-converting ability of recombinant Saccharomyces cerevisiae SX6(MUT) expressing NADH-preferring xylose reductase mutant (R276H) and other xylose-metabolic enzymes, and deficient in aldehyde dehydrogenase 6 (Ald6p) were characterized at microaerobic conditions using various sugar mixtures. The reduction of air supply from 0.5vvm to 0.1vvm increased specific ethanol production rate by 75% and did not affect specific xylose consumption rate. In batch fermentations using various concentrations of xylose (50-104g/L), higher xylose concentration enhanced xylose consumption rate and ethanol productivity but reduced ethanol yield, owing to the accumulation of xylitol and glycerol from xylose. SX6(MUT) consumed monosugars in pitch pine hydrolysates and produced 23.1g/L ethanol from 58.7g/L sugars with 0.39g/g ethanol yield, which was 14% higher than the host strain of S. cerevisiae D452-2 without the xylose assimilating enzymes. In conclusion, S. cerevisiae SX6(MUT) was characterized to possess high xylose-consuming ability in microaerobic conditions and a potential for ethanol production from cellulosic biomass. Copyright © 2016 Elsevier B.V. All rights reserved.

Establishment of oxidative D-xylose metabolism in Pseudomonas putida S12

NARCIS (Netherlands)

Meijnen, J.P.; Winde, J.H. de; Ruijssenaars, H.J.

2009-01-01

The oxidative D-xylose catabolic pathway of Caulobacter crescentus, encoded by the xylXABCD operon, was expressed in the gram-negative bacterium Pseudomonas putida S12. This engineered transformant strain was able to grow on D-xylose as a sole carbon source with a biomass yield of 53% (based on g

Separate and Simultaneous enzymatic hydrolysis and fermentation of wheat hemicellulose with recombinant xylose utilizing Saccharomyces cerevisiae

DEFF Research Database (Denmark)

Olsson, Lisbeth; Sørensen, H. R.; Dam, B. P

2006-01-01

Fermentations with three different xylose-utilizing recombinant Saccharomyces cerevisiae strains (F12, CR4, and CB4) were performed using two different wheat hemicellulose substrates, unfermented starch free fibers, and an industrial ethanol fermentation residue, vinasse. With CR4 and F12......, the maximum ethanol concentrations obtained were 4.3 and 4 g/L, respectively, but F12 converted xylose 15% faster than CR4 during the first 24 h. The comparison of separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) with F12 showed that the highest, maximum...... ethanol concentrations were obtained with SSF. In general, the volumetric ethanol productivity was initially, highest in the SHF, but the overall volumetric ethanol productivity ended up being maximal in the SSF, at 0.013 and 0.010 g/Lh, with starch free fibers and vinasse, respectively....

Pichia stipitis xylose reductase helps detoxifying lignocellulosic hydrolysate by reducing 5-hydroxymethyl-furfural (HMF

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Röder Anja

2008-06-01

Full Text Available Abstract Background Pichia stipitis xylose reductase (Ps-XR has been used to design Saccharomyces cerevisiae strains that are able to ferment xylose. One example is the industrial S. cerevisiae xylose-consuming strain TMB3400, which was constructed by expression of P. stipitis xylose reductase and xylitol dehydrogenase and overexpression of endogenous xylulose kinase in the industrial S. cerevisiae strain USM21. Results In this study, we demonstrate that strain TMB3400 not only converts xylose, but also displays higher tolerance to lignocellulosic hydrolysate during anaerobic batch fermentation as well as 3 times higher in vitro HMF and furfural reduction activity than the control strain USM21. Using laboratory strains producing various levels of Ps-XR, we confirm that Ps-XR is able to reduce HMF both in vitro and in vivo. Ps-XR overexpression increases the in vivo HMF conversion rate by approximately 20%, thereby improving yeast tolerance towards HMF. Further purification of Ps-XR shows that HMF is a substrate inhibitor of the enzyme. Conclusion We demonstrate for the first time that xylose reductase is also able to reduce the furaldehyde compounds that are present in undetoxified lignocellulosic hydrolysates. Possible implications of this newly characterized activity of Ps-XR on lignocellulosic hydrolysate fermentation are discussed.

Ethanol production in fermentation of mixed sugars containing xylose

Science.gov (United States)

Viitanen, Paul V [West Chester, PA; Mc Cutchen, Carol M [Wilmington, DE; Li,; Xu, [Newark, DE; Emptage, Mark [Wilmington, DE; Caimi, Perry G [Kennett Square, PA; Zhang, Min [Lakewood, CO; Chou, Yat-Chen [Lakewood, CO; Franden, Mary Ann [Centennial, CO

2009-12-08

Xylose-utilizing Z. mobilis strains were found to have improved ethanol production when grown in medium containing mixed sugars including xylose if sorbitol or mannitol was included in the medium. The effect was seen in concentrations of mixed sugars where no growth lag period occurs, as well as in higher sugars concentrations.

Production of Xylitol from D-Xylose by Overexpression of Xylose Reductase in Osmotolerant Yeast Candida glycerinogenes WL2002-5.

Science.gov (United States)

Zhang, Cheng; Zong, Hong; Zhuge, Bin; Lu, Xinyao; Fang, Huiying; Zhuge, Jian

2015-07-01

Efficient bioconversion of D-xylose into various biochemicals is critical for the developing lignocelluloses application. In this study, we compared D-xylose utilization in Candida glycerinogenes WL2002-5 transformants expressing xylose reductase (XYL1) in D-xylose metabolism. C. glycerinogenes WL2002-5 expressing XYL1 from Schefferomyces stipitis can produce xylitol. Xylitol production by the recombinant strains was evaluated using a xylitol fermentation medium with glucose as a co-substrate. As glucose was found to be an insufficient co-substrate, various carbon sources were screened for efficient cofactor regeneration, and glycerol was found to be the best co-substrate. The effects of glycerol on the xylitol production rate by a xylose reductase gene (XYL1)-overexpressed mutant of C. glycerinogenes WL2002-5 were investigated. The XYL1-overexpressed mutant produced xylitol from D-xylose using glycerol as a co-substrate for cell growth and NAD (P) H regeneration: 100 g/L D-xylose was completely converted into xylitol when at least 20 g/L glycerol was used as a co-substrate. XYL1 overexpressed mutant grown on glycerol as co-substrate accumulated 2.1-fold increased xylitol concentration over those cells grown on glucose as co-substrate. XYL1 overexpressed mutant produced xylitol with a volumetric productivity of 0.83 g/L/h, and a xylitol yield of 98 % xylose. Recombinant yeast strains obtained in this study are promising candidates for xylitol production. This is the first report of XYL1 gene overexpression of C. glycerinogenes WL2002-5 for enhancing the efficiency of xylitol production.

Impact of overexpressing NADH kinase on glucose and xylose metabolism in recombinant xylose-utilizing Saccharomyces cerevisiae

DEFF Research Database (Denmark)

Hou, Jin; Vemuri, G. N.; Bao, X. M.

2009-01-01

of overexpressing the native NADH kinase (encoded by the POS5 gene) in xylose-consuming recombinant S. cerevisiae directed either into the cytosol or to the mitochondria was evaluated. The physiology of the NADH kinase containing strains was also evaluated during growth on glucose. Overexpressing NADH kinase...

Process for assembly and transformation into Saccharomyces cerevisiae of a synthetic yeast artificial chromosome containing a multigene cassette to express enzymes that enhance xylose utilization designed for an automated pla

Science.gov (United States)

A yeast artificial chromosome (YAC) containing a multigene cassette for expression of enzymes that enhance xylose utilization (xylose isomerase [XI] and xylulokinase [XKS]) was constructed and transformed into Saccharomyces cerevisiae to demonstrate feasibility as a stable protein expression system ...

Isolation and characterization of a sulfur-oxidizing chemolithotroph growing on crude oil under anaerobic conditions.

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Kodama, Yumiko; Watanabe, Kazuya

2003-01-01

Molecular approaches have shown that a group of bacteria (called cluster 1 bacteria) affiliated with the epsilon subclass of the class Proteobacteria constituted major populations in underground crude-oil storage cavities. In order to unveil their physiology and ecological niche, this study isolated bacterial strains (exemplified by strain YK-1) affiliated with the cluster 1 bacteria from an oil storage cavity at Kuji in Iwate, Japan. 16S rRNA gene sequence analysis indicated that its closest relative was Thiomicrospira denitrificans (90% identity). Growth experiments under anaerobic conditions showed that strain YK-1 was a sulfur-oxidizing obligate chemolithotroph utilizing sulfide, elemental sulfur, thiosulfate, and hydrogen as electron donors and nitrate as an electron acceptor. Oxygen also supported its growth only under microaerobic conditions. Strain YK-1 could not grow on nitrite, and nitrite was the final product of nitrate reduction. Neither sugars, organic acids (including acetate), nor hydrocarbons could serve as carbon and energy sources. A typical stoichiometry of its energy metabolism followed an equation: S(2-) + 4NO(3)(-) --> SO(4)(2-) + 4NO(2)(-) (Delta G(0) = -534 kJ mol(-1)). In a difference from other anaerobic sulfur-oxidizing bacteria, this bacterium was sensitive to NaCl; growth in medium containing more than 1% NaCl was negligible. When YK-1 was grown anaerobically in a sulfur-depleted inorganic medium overlaid with crude oil, sulfate was produced, corresponding to its growth. On the contrary, YK-1 could not utilize crude oil as a carbon source. These results suggest that the cluster 1 bacteria yielded energy for growth in oil storage cavities by oxidizing petroleum sulfur compounds. Based on its physiology, ecological interactions with other members of the groundwater community are discussed.

Effects of NADH-preferring xylose reductase expression on ethanol production from xylose in xylose-metabolizing recombinant Saccharomyces cerevisiae.

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Lee, Sung-Haeng; Kodaki, Tsutomu; Park, Yong-Cheol; Seo, Jin-Ho

2012-04-30

Efficient conversion of xylose to ethanol is an essential factor for commercialization of lignocellulosic ethanol. To minimize production of xylitol, a major by-product in xylose metabolism and concomitantly improve ethanol production, Saccharomyces cerevisiae D452-2 was engineered to overexpress NADH-preferable xylose reductase mutant (XR(MUT)) and NAD⁺-dependent xylitol dehydrogenase (XDH) from Pichia stipitis and endogenous xylulokinase (XK). In vitro enzyme assay confirmed the functional expression of XR(MUT), XDH and XK in recombinant S. cerevisiae strains. The change of wild type XR to XR(MUT) along with XK overexpression led to reduction of xylitol accumulation in microaerobic culture. More modulation of the xylose metabolism including overexpression of XR(MUT) and transaldolase, and disruption of the chromosomal ALD6 gene encoding aldehyde dehydrogenase (SX6(MUT)) improved the performance of ethanol production from xylose remarkably. Finally, oxygen-limited fermentation of S. cerevisiae SX6(MUT) resulted in 0.64 g l⁻¹ h⁻¹ xylose consumption rate, 0.25 g l⁻¹ h⁻¹ ethanol productivity and 39% ethanol yield based on the xylose consumed, which were 1.8, 4.2 and 2.2 times higher than the corresponding values of recombinant S. cerevisiae expressing XR(MUT), XDH and XK only. Copyright © 2011 Elsevier B.V. All rights reserved.

Alcoholic glucose and xylose fermentations by the coculture process: Compatability and typing of associated strains

Energy Technology Data Exchange (ETDEWEB)

Laplace, J.M.; Delgenes, J.P.; Moletta, R. (Institut national de la recherche agronomique, Narbonne (France)); Navarro, J.M. (Universite de Montpellier (France))

1992-01-01

As part of the simulaneous fermentation of both glucose and xylose to ethanol by a coculture process, compatibilities between xylose-fermenting yeasts and glucose-fermenting species were investigated. Among the Saccharomyces species tested, none inhibited growth of the xylose-fermenting yeasts. By contrast, many xylose-fermenting yeasts, among the 11 tested, exerted an inhibitory effect on growth of the selected Saccharomyces species. Killer character was demonstrated in three strains of Pichia stipitis. Such strains, despite their high fermentative performances, cannot be used to ferment D-xylose in association with the selected Saccharomyces species. From compatibility tests between xylose-fermenting yeasts and Saccharomyces species, pairs of microorganisms suitable for simultaneous xylose and glucose fermentations by coculture are proposed. Strains associated in the coculture process are distinguished by their resistance to mitochondrial inhibitors. The xylose-fermenting yeasts are able to grow on media containing erythromycin (1 g/l) or diuron (50 mg/l), whereas, the Saccharomyces species are inhibited by these mitochondrial inhibitors. 15 refs., 2 figs., 3 tabs.

Single zymomonas mobilis strain for xylose and arabinose fermentation

Science.gov (United States)

Zhang, Min; Chou, Yat-Chen; Picataggio, Stephen K.; Finkelstein, Mark

1998-01-01

This invention relates to single microorganisms which normally do not ferment pentose sugars which are genetically altered to ferment the pentose sugars, xylose and arabinose, to produce ethanol, and a fermentation process utilizing the same. Examples include Zymomonas mobilis which has been transformed with a combination of E. coli genes for xylose isomerase, xylulokinase, L-arabinose isomerase, L-ribulokinase, L-ribulose 5-phosphate 4-epimerase, transaldolase and transketolase. Expression of added genes are under the control of Z. mobilis promoters. These newly created microorganisms are useful for fermenting glucose, xylose and arabinose, produced by hydrolysis of hemicellulose and cellulose or starch, to produce ethanol.

Intrinsic kinetic parameters of substrate utilization by immobilized anaerobic sludge.

Science.gov (United States)

Zaiat, M; Vieira, L G; Foresti, E

1997-01-20

This article presents a method for evaluating the intrinsic kinetic parameters of the specific substrate utilization rate (r) equation and discusses the results obtained for anaerobic sludge-bed samples taken from a horizontal-flow anaerobic immobilized sludge (HAIS) reactor. This method utilizes a differential reactor filled with polyurethane foam matrices containing immobilized anaerobic sludge which is subjected to a range of feeding substrate flow rates. The range of liquid superficial velocities thus obtained are used for generating data of observed specific substrate utilization rates (r(obs)) under a diversity of external mass transfer resistance conditions. The r(obs) curves are then adjusted to permit their extrapolation for the condition of no external mass transfer resistance, and the values determined are used as a test for the condition of absence of limitation of internal mass transfer. The intrinsic parameters r(max), the maximum specific substrate utilization rate, and K(s), the half-velocity coefficient, are evaluated from the r values under no external mass transfer resistance and no internal mass transfer limitation. The application of such a method for anaerobic sludge immobilized in polyurethane foam particles treating a glucose substrate at 30 degrees C resulted in intrinsic r(max) and K(s), respectively, of 0.330 mg chemical oxygen demand (COD) . mg(-1) volatile suspended solids (VSS) . h(-1) and 72 mg COD . L(-1). In comparison with the values found in the literature, intrinsic r(max) is significantly high and intrinsic K(s) is relatively low. (c) 1997 John Wiley & Sons, Inc.

Ethanol production from lignocellulosic hydrolysates using engineered Saccharomyces cerevisiae harboring xylose isomerase-based pathway.

Science.gov (United States)

Ko, Ja Kyong; Um, Youngsoon; Woo, Han Min; Kim, Kyoung Heon; Lee, Sun-Mi

2016-06-01

The efficient co-fermentation of glucose and xylose is necessary for the economically feasible bioethanol production from lignocellulosic biomass. Even with xylose utilizing Saccharomyces cerevisiae, the efficiency of the lignocellulosic ethanol production remains suboptimal mainly due to the low conversion yield of xylose to ethanol. In this study, we evaluated the co-fermentation performances of SXA-R2P-E, a recently engineered isomerase-based xylose utilizing strain, in mixed sugars and in lignocellulosic hydrolysates. In a high-sugar fermentation with 70g/L of glucose and 40g/L of xylose, SXA-R2P-E produced 50g/L of ethanol with an yield of 0.43gethanol/gsugars at 72h. From dilute acid-pretreated hydrolysates of rice straw and hardwood (oak), the strain produced 18-21g/L of ethanol with among the highest yield of 0.43-0.46gethanol/gsugars ever reported. This study shows a highly promising potential of a xylose isomerase-expressing strain as an industrially relevant ethanol producer from lignocellulosic hydrolysates. Copyright © 2016 Elsevier Ltd. All rights reserved.

An efficient xylose-fermenting recombinant Saccharomyces cerevisiae strain obtained through adaptive evolution and its global transcription profile

Energy Technology Data Exchange (ETDEWEB)

Shen, Yu; Chen, Xiao; Peng, Bingyin; Chen, Liyuan; Hou, Jin; Bao, Xiaoming [Shandong Univ., Jinan (China). State Key Lab. of Microbial Technology

2012-11-15

Factors related to ethanol production from xylose in engineered Saccharomyces cerevisiae that contain an exogenous initial metabolic pathway are still to be elucidated. In the present study, a strain that expresses the xylose isomerase gene of Piromyces sp. Pi-xylA and overexpresses XKS1, RPE1, RKI1, TAL1, and TKL1, with deleted GRE3 and COX4 genes was constructed. The xylose utilization capacity of the respiratory deficiency strain was poor but improved via adaptive evolution in xylose. The {mu}{sub max} of the evolved strain in 20 gl{sup -1} xylose is 0.11 {+-} 0.00 h{sup -1}, and the evolved strain consumed 17.83 gl{sup -1} xylose within 72 h, with an ethanol yield of 0.43 gg{sup -1} total consumed sugars during glucose-xylose cofermentation. Global transcriptional changes and effect of several specific genes were studied. The result revealed that the increased xylose isomerase activity, the upregulation of enzymes involved in glycolysis and glutamate synthesis, and the downregulation of trehalose and glycogen synthesis, may have contributed to the improved xylose utilization of the strain. Furthermore, the deletion of PHO13 decreased the xylose growth in the respiration deficiency strain although deleting PHO13 can improve the xylose metabolism in other strains. (orig.)

Genetic analysis of D-xylose metabolism by endophytic yeast strains of Rhodotorula graminis and Rhodotorula mucilaginosa

Directory of Open Access Journals (Sweden)

Ping Xu

2011-01-01

Full Text Available Two novel endophytic yeast strains, WP1 and PTD3, isolated from within the stems of poplar (Populus trees, were genetically characterized with respect to their xylose metabolism genes. These two strains, belonging to the species Rhodotorula graminis and R. mucilaginosa, respectively, utilize both hexose and pentose sugars, including the common plant pentose sugar, D-xylose. The xylose reductase (XYL1 and xylitol dehydrogenase (XYL2 genes were cloned and characterized. The derived amino acid sequences of xylose reductase (XR and xylose dehydrogenase (XDH were 32%~41% homologous to those of Pichia stipitis and Candida. spp., two species known to utilize xylose. The derived XR and XDH sequences of WP1 and PTD3 had higher homology (73% and 69% identity with each other. WP1 and PTD3 were grown in single sugar and mixed sugar media to analyze the XYL1 and XYL2 gene regulation mechanisms. Our results revealed that for both strains, the gene expression is induced by D-xylose, and that in PTD3 the expression was not repressed by glucose in the presence of xylose.

Production of Bioethanol From Lignocellulosic Biomass Using Thermophilic Anaerobic Bacteria

DEFF Research Database (Denmark)

Georgieva, Tania I.

2006-01-01

and xylose and to tolerate the inhibitory compounds present in lignocellulosic hydrolysates is therefore apparent. Several thermophilic anaerobic xylan degrading bacteria from our culture collection (EMB group at BioCentrum-DTU) have been screened for a potential ethanol producer from hemicellulose...... hydrolysates, and out of the screening test, one particular strain (A10) was selected for the best performance. The strain was morphologically and physiologically characterized as Thermoanaerobacter mathranii strain A10. Unlike other thermophilic anaerobic bacteria, the wild-type strain Thermoanaerobacter...... Thermoanaerobacter BG1L1 was further studied. The experiments were carried out in a continuous immobilized reactor system (a fluidized bed reactor), which is likely to be the process design configuration for xylose fermentation in a Danish biorefinery concept for production of fuel ethanol. The immobilization...

Mutations in iron-sulfur cluster proteins that improve xylose utilization

Science.gov (United States)

Froehlich, Allan; Henningsen, Brooks; Covalla, Sean; Zelle, Rintze M.

2018-03-20

There is provided an engineered host cells comprising (a) one or more mutations in one or more endogenous genes encoding a protein associated with iron metabolism; and (b) at least one gene encoding a polypeptide having xylose isomerase activity, and methods of their use thereof.

Application of Methanobrevibacter acididurans in anaerobic digestion.

Science.gov (United States)

Savant, D V; Ranade, D R

2004-01-01

To operate anaerobic digesters successfully under acidic conditions, hydrogen utilizing methanogens which can grow efficiently at low pH and tolerate high volatile fatty acids (VFA) are desirable. An acid tolerant hydrogenotrophic methanogen viz. Methanobrevibacter acididurans isolated from slurry of an anaerobic digester running on alcohol distillery wastewater has been described earlier by this lab. This organism could grow optimally at pH 6.0. In the experiments reported herein, M. acididurans showed better methanogenesis under acidic conditions with high VFA, particularly acetate, than Methanobacterium bryantii, a common hydrogenotrophic inhabitant of anaerobic digesters. Addition of M. acididurans culture to digesting slurry of acidogenic as well as methanogenic digesters running on distillery wastewater showed increase in methane production and decrease in accumulation of volatile fatty acids. The results proved the feasibility of application of M. acididurans in anaerobic digesters.

Engineering of xylose reductase and overexpression of xylitol dehydrogenase and xylulokinase improves xylose alcoholic fermentation in the thermotolerant yeast Hansenula polymorpha

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Voronovsky Andriy Y

2008-07-01

Full Text Available Abstract Background The thermotolerant methylotrophic yeast Hansenula polymorpha is capable of alcoholic fermentation of xylose at elevated temperatures (45 – 48°C. Such property of this yeast defines it as a good candidate for the development of an efficient process for simultaneous saccharification and fermentation. However, to be economically viable, the main characteristics of xylose fermentation of H. polymorpha have to be improved. Results Site-specific mutagenesis of H. polymorpha XYL1 gene encoding xylose reductase was carried out to decrease affinity of this enzyme toward NADPH. The modified version of XYL1 gene under control of the strong constitutive HpGAP promoter was overexpressed on a Δxyl1 background. This resulted in significant increase in the KM for NADPH in the mutated xylose reductase (K341 → R N343 → D, while KM for NADH remained nearly unchanged. The recombinant H. polymorpha strain overexpressing the mutated enzyme together with native xylitol dehydrogenase and xylulokinase on Δxyl1 background was constructed. Xylose consumption, ethanol and xylitol production by the constructed strain were determined for high-temperature xylose fermentation at 48°C. A significant increase in ethanol productivity (up to 7.3 times was shown in this recombinant strain as compared with the wild type strain. Moreover, the xylitol production by the recombinant strain was reduced considerably to 0.9 mg × (L × h-1 as compared to 4.2 mg × (L × h-1 for the wild type strain. Conclusion Recombinant strains of H. polymorpha engineered for improved xylose utilization are described in the present work. These strains show a significant increase in ethanol productivity with simultaneous reduction in the production of xylitol during high-temperature xylose fermentation.

Impact of zinc supplementation on the improved fructose/xylose utilization and butanol production during acetone-butanol-ethanol fermentation.

Science.gov (United States)

Wu, You-Duo; Xue, Chuang; Chen, Li-Jie; Bai, Feng-Wu

2016-01-01

Lignocellulosic biomass and dedicated energy crops such as Jerusalem artichoke are promising alternatives for biobutanol production by solventogenic clostridia. However, fermentable sugars such as fructose or xylose released from the hydrolysis of these feedstocks were subjected to the incomplete utilization by the strains, leading to relatively low butanol production and productivity. When 0.001 g/L ZnSO4·7H2O was supplemented into the medium containing fructose as sole carbon source, 12.8 g/L of butanol was achieved with butanol productivity of 0.089 g/L/h compared to only 4.5 g/L of butanol produced with butanol productivity of 0.028 g/L/h in the control without zinc supplementation. Micronutrient zinc also led to the improved butanol production up to 8.3 g/L derived from 45.2 g/L xylose as sole carbon source with increasing butanol productivity by 31.7%. Moreover, the decreased acids production was observed under the zinc supplementation condition, resulting in the increased butanol yields of 0.202 g/g-fructose and 0.184 g/g-xylose, respectively. Similar improvements were also observed with increasing butanol production by 130.2 % and 8.5 %, butanol productivity by 203.4% and 18.4%, respectively, in acetone-butanol-ethanol fermentations from sugar mixtures of fructose/glucose (4:1) and xylose/glucose (1:2) simulating the hydrolysates of Jerusalem artichoke tubers and corn stover. The results obtained from transcriptional analysis revealed that zinc may have regulatory mechanisms for the sugar transport and metabolism of Clostridium acetobutylicum L7. Therefore, micronutrient zinc supplementation could be an effective way for economic development of butanol production derived from these low-cost agricultural feedstocks. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Increased ethanol production by deletion of HAP4 in recombinant xylose-assimilating Saccharomyces cerevisiae.

Science.gov (United States)

Matsushika, Akinori; Hoshino, Tamotsu

2015-12-01

The Saccharomyces cerevisiae HAP4 gene encodes a transcription activator that plays a key role in controlling the expression of genes involved in mitochondrial respiration and reductive pathways. This work examines the effect of knockout of the HAP4 gene on aerobic ethanol production in a xylose-utilizing S. cerevisiae strain. A hap4-deleted recombinant yeast strain (B42-DHAP4) showed increased maximum concentration, production rate, and yield of ethanol compared with the reference strain MA-B42, irrespective of cultivation medium (glucose, xylose, or glucose/xylose mixtures). Notably, B42-DHAP4 was capable of producing ethanol from xylose as the sole carbon source under aerobic conditions, whereas no ethanol was produced by MA-B42. Moreover, the rate of ethanol production and ethanol yield (0.44 g/g) from the detoxified hydrolysate of wood chips was markedly improved in B42-DHAP4 compared to MA-B42. Thus, the results of this study support the view that deleting HAP4 in xylose-utilizing S. cerevisiae strains represents a useful strategy in ethanol production processes.

Improved Xylose Metabolism by a CYC8 Mutant of Saccharomyces cerevisiae.

Science.gov (United States)

Nijland, Jeroen G; Shin, Hyun Yong; Boender, Leonie G M; de Waal, Paul P; Klaassen, Paul; Driessen, Arnold J M

2017-06-01

Engineering Saccharomyces cerevisiae for the utilization of pentose sugars is an important goal for the production of second-generation bioethanol and biochemicals. However, S. cerevisiae lacks specific pentose transporters, and in the presence of glucose, pentoses enter the cell inefficiently via endogenous hexose transporters (HXTs). By means of in vivo engineering, we have developed a quadruple hexokinase deletion mutant of S. cerevisiae that evolved into a strain that efficiently utilizes d-xylose in the presence of high d-glucose concentrations. A genome sequence analysis revealed a mutation (Y353C) in the general corepressor CYC8 , or SSN6 , which was found to be responsible for the phenotype when introduced individually in the nonevolved strain. A transcriptome analysis revealed altered expression of 95 genes in total, including genes involved in (i) hexose transport, (ii) maltose metabolism, (iii) cell wall function (mannoprotein family), and (iv) unknown functions (seripauperin multigene family). Of the 18 known HXTs, genes for 9 were upregulated, especially the low or nonexpressed HXT10 , HXT13 , HXT15 , and HXT16 Mutant cells showed increased uptake rates of d-xylose in the presence of d-glucose, as well as elevated maximum rates of metabolism ( V max ) for both d-glucose and d-xylose transport. The data suggest that the increased expression of multiple hexose transporters renders d-xylose metabolism less sensitive to d-glucose inhibition due to an elevated transport rate of d-xylose into the cell. IMPORTANCE The yeast Saccharomyces cerevisiae is used for second-generation bioethanol formation. However, growth on xylose is limited by pentose transport through the endogenous hexose transporters (HXTs), as uptake is outcompeted by the preferred substrate, glucose. Mutant strains were obtained with improved growth characteristics on xylose in the presence of glucose, and the mutations mapped to the regulator Cyc8. The inactivation of Cyc8 caused increased

Biological hydrogen production by moderately thermophilic anaerobic bacteria

International Nuclear Information System (INIS)

HP Goorissen; AJM Stams

2006-01-01

This study focuses on the biological production of hydrogen at moderate temperatures (65-75 C) by anaerobic bacteria. A survey was made to select the best (moderate) thermophiles for hydrogen production from cellulolytic biomass. From this survey we selected Caldicellulosiruptor saccharolyticus (a gram-positive bacterium) and Thermotoga elfii (a gram-negative bacterium) as potential candidates for biological hydrogen production on mixtures of C 5 -C 6 sugars. Xylose and glucose were used as model substrates to describe growth and hydrogen production from hydrolyzed biomass. Mixed substrate utilization in batch cultures revealed differences in the sequence of substrate consumption and in catabolites repression of the two microorganisms. The regulatory mechanisms of catabolites repression in these microorganisms are not known yet. (authors)

Xylose fermentation to ethanol

Energy Technology Data Exchange (ETDEWEB)

McMillan, J.D.

1993-01-01

The past several years have seen tremendous progress in the understanding of xylose metabolism and in the identification, characterization, and development of strains with improved xylose fermentation characteristics. A survey of the numerous microorganisms capable of directly fermenting xylose to ethanol indicates that wild-type yeast and recombinant bacteria offer the best overall performance in terms of high yield, final ethanol concentration, and volumetric productivity. The best performing bacteria, yeast, and fungi can achieve yields greater than 0.4 g/g and final ethanol concentrations approaching 5%. Productivities remain low for most yeast and particularly for fungi, but volumetric productivities exceeding 1.0 g/L-h have been reported for xylose-fermenting bacteria. In terms of wild-type microorganisms, strains of the yeast Pichia stipitis show the most promise in the short term for direct high-yield fermentation of xylose without byproduct formation. Of the recombinant xylose-fermenting microorganisms developed, recombinant E. coli ATTC 11303 (pLOI297) exhibits the most favorable performance characteristics reported to date.

D-xylose absorption

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... this page: //medlineplus.gov/ency/article/003606.htm D-xylose absorption To use the sharing features on this page, please enable JavaScript. D-xylose absorption is a laboratory test to determine ...

An evolved xylose transporter from Zymomonas mobilis enhances sugar transport in Escherichia coli

Directory of Open Access Journals (Sweden)

Zhang Jingqing

2009-12-01

Full Text Available Abstract Background Xylose is a second most abundant sugar component of lignocellulose besides glucose. Efficient fermentation of xylose is important for the economics of biomass-based biorefineries. However, sugar mixtures are sequentially consumed in xylose co-fermentation with glucose due to carbon catabolite repression (CCR in microorganisms. As xylose transmembrance transport is one of the steps repressed by CCR, it is therefore of interest to develop a transporter that is less sensitive to the glucose inhibition or CCR. Results The glucose facilitator protein Glf transporter from Zymomonas mobilis, also an efficient transporter for xylose, was chosen as the target transporter for engineering to eliminate glucose inhibition on xylose uptake. The evolution of Glf transporter was carried out with a mixture of glucose and xylose in E. coli. Error-prone PCR and random deletion were employed respectively in two rounds of evolution. Aided by a high-throughput screening assay using xylose analog p-nitrophenyl-β-D-xylopyranoside (pNPX in 96-well plates, a best mutant 2-RD5 was obtained that contains several mutations, and a deletion of 134 residues (about 28% of total residues, or three fewer transmembrane sections (TMSs. It showed a 10.8-fold improvement in terms of pNPX transport activity in the presence of glucose. The fermentation performance results showed that this mutant improved xylose consumption by 42% with M9 minimal medium containing 20 g L-1 xylose only, while with the mixture sugar of xylose and glucose, 28% more glucose was consumed, but no obvious co-utilization of xylose was observed. Further glucose fed-batch experiments suggested that the intracellular metabolism of xylose was repressed by glucose. Conclusions Through random mutagenesis and partial deletion coupled with high-throughput screening, a mutant of the Glf transporter (2-RD5 was obtained that relieved the inhibition of xylose transport by glucose. The fermentation

The effect of initial cell concentration on xylose fermentation by Pichia stipitis

Science.gov (United States)

Frank K. Agbogbo; Guillermo Coward-Kelly; Mads Torry-Smith; Kevin Wenger; Thomas W. Jeffries

2007-01-01

Xylose was fermented using Pichia stipitis CBS 6054 at different initial cell concentrations. A high initial cell concentration increased the rate of xylose utilization, ethanol formation, and the ethanol yield. The highest ethanol concentration of 41.0 g/L and a yield of 0.38 g/g was obtained using an initial cell concentration of 6.5 g/L. Even though more xylitol was...

Metabolic Engineering of Escherichia coli K12 for Homofermentative Production of L-Lactate from Xylose.

Science.gov (United States)

Jiang, Ting; Zhang, Chen; He, Qin; Zheng, Zhaojuan; Ouyang, Jia

2018-02-01

The efficient utilization of xylose is regarded as a technical barrier to the commercial production of bulk chemicals from biomass. Due to the desirable mechanical properties of polylactic acid (PLA) depending on the isomeric composition of lactate, biotechnological production of lactate with high optical pure has been increasingly focused in recent years. The main objective of this work was to construct an engineered Escherichia coli for the optically pure L-lactate production from xylose. Six chromosomal deletions (pflB, ldhA, ackA, pta, frdA, adhE) and a chromosomal integration of L-lactate dehydrogenase-encoding gene (ldhL) from Bacillus coagulans was involved in construction of E. coli KSJ316. The recombinant strain could produce L-lactate from xylose resulting in a yield of 0.91 g/g xylose. The chemical purity of L-lactate was 95.52%, and the optical purity was greater than 99%. Moreover, three strategies, including overexpression of L-lactate dehydrogenase, intensification of xylose catabolism, and addition of additives to medium, were designed to enhance the production. The results showed that they could increase the concentration of L-lactate by 32.90, 20.13, and 233.88% relative to the control, respectively. This was the first report that adding formate not only could increase the xylose utilization but also led to the fewer by-product levels.

Genomic analysis of a xylose operon and characterization of novel xylose isomerase and xylulokinase from Bacillus coagulans NL01.

Science.gov (United States)

Zheng, Zhaojuan; Lin, Xi; Jiang, Ting; Ye, Weihua; Ouyang, Jia

2016-08-01

To investigate the xylose operon and properties of xylose isomerase and xylulokinase in Bacillus coagulans that can effectively ferment xylose to lactic acid. The xylose operon is widely present in B. coagulans. It is composed of four putative ORFs. Novel xylA and xylB from B. coagulans NL01 were cloned and expressed in Escherichia coli. Sequence of xylose isomerase was more conserved than that of xylulokinase. Both the enzymes exhibited maximum activities at pH 7-8 but with a high temperature maximum of 80-85 °C, divalent metal ion was prerequisite for their activation. Xylose isomerase and xylulokinase were most effectively activated by Ni(2+) and Co(2+), respectively. Genomic analysis of xylose operon has contributed to understanding xylose metabolism in B. coagulans and the novel xylose isomerase and xylulokinase might provide new alternatives for metabolic engineering of other strains to improve their fermentation performance on xylose.

Time-based comparative transcriptomics in engineered xylose-utilizing Saccharomyces cerevisiae identifies temperature-responsive genes during ethanol production.

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Ismail, Ku Syahidah Ku; Sakamoto, Takatoshi; Hasunuma, Tomohisa; Kondo, Akihiko

2013-09-01

Agricultural residues comprising lignocellulosic materials are excellent sources of pentose sugar, which can be converted to ethanol as fuel. Ethanol production via consolidated bioprocessing requires a suitable microorganism to withstand the harsh fermentation environment of high temperature, high ethanol concentration, and exposure to inhibitors. We genetically enhanced an industrial Saccharomyces cerevisiae strain, sun049, enabling it to uptake xylose as the sole carbon source at high fermentation temperature. This strain was able to produce 13.9 g/l ethanol from 50 g/l xylose at 38 °C. To better understand the xylose consumption ability during long-term, high-temperature conditions, we compared by transcriptomics two fermentation conditions: high temperature (38 °C) and control temperature (30 °C) during the first 12 h of fermentation. This is the first long-term, time-based transcriptomics approach, and it allowed us to discover the role of heat-responsive genes when xylose is the sole carbon source. The results suggest that genes related to amino acid, cell wall, and ribosomal protein synthesis are down-regulated under heat stress. To allow cell stability and continuous xylose uptake in order to produce ethanol, hexose transporter HXT5, heat shock proteins, ubiquitin proteins, and proteolysis were all induced at high temperature. We also speculate that the strong relationship between high temperature and increased xylitol accumulation represents the cell's mechanism to protect itself from heat degradation.

Xylose isomerase improves growth and ethanol production rates from biomass sugars for both Saccharomyces pastorianus and Saccharomyces cerevisiae.

Science.gov (United States)

Miller, Kristen P; Gowtham, Yogender Kumar; Henson, J Michael; Harcum, Sarah W

2012-01-01

The demand for biofuel ethanol made from clean, renewable nonfood sources is growing. Cellulosic biomass, such as switch grass (Panicum virgatum L.), is an alternative feedstock for ethanol production; however, cellulosic feedstock hydrolysates contain high levels of xylose, which needs to be converted to ethanol to meet economic feasibility. In this study, the effects of xylose isomerase on cell growth and ethanol production from biomass sugars representative of switch grass were investigated using low cell density cultures. The lager yeast species Saccharomyces pastorianus was grown with immobilized xylose isomerase in the fermentation step to determine the impact of the glucose and xylose concentrations on the ethanol production rates. Ethanol production rates were improved due to xylose isomerase; however, the positive effect was not due solely to the conversion of xylose to xylulose. Xylose isomerase also has glucose isomerase activity, so to better understand the impact of the xylose isomerase on S. pastorianus, growth and ethanol production were examined in cultures provided fructose as the sole carbon. It was observed that growth and ethanol production rates were higher for the fructose cultures with xylose isomerase even in the absence of xylose. To determine whether the positive effects of xylose isomerase extended to other yeast species, a side-by-side comparison of S. pastorianus and Saccharomyces cerevisiae was conducted. These comparisons demonstrated that the xylose isomerase increased ethanol productivity for both the yeast species by increasing the glucose consumption rate. These results suggest that xylose isomerase can contribute to improved ethanol productivity, even without significant xylose conversion. Copyright © 2012 American Institute of Chemical Engineers (AIChE).

Reconstruction and analysis of a genome-scale metabolic model for Scheffersomyces stipitis

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Balagurunathan Balaji

2012-02-01

Full Text Available Abstract Background Fermentation of xylose, the major component in hemicellulose, is essential for economic conversion of lignocellulosic biomass to fuels and chemicals. The yeast Scheffersomyces stipitis (formerly known as Pichia stipitis has the highest known native capacity for xylose fermentation and possesses several genes for lignocellulose bioconversion in its genome. Understanding the metabolism of this yeast at a global scale, by reconstructing the genome scale metabolic model, is essential for manipulating its metabolic capabilities and for successful transfer of its capabilities to other industrial microbes. Results We present a genome-scale metabolic model for Scheffersomyces stipitis, a native xylose utilizing yeast. The model was reconstructed based on genome sequence annotation, detailed experimental investigation and known yeast physiology. Macromolecular composition of Scheffersomyces stipitis biomass was estimated experimentally and its ability to grow on different carbon, nitrogen, sulphur and phosphorus sources was determined by phenotype microarrays. The compartmentalized model, developed based on an iterative procedure, accounted for 814 genes, 1371 reactions, and 971 metabolites. In silico computed growth rates were compared with high-throughput phenotyping data and the model could predict the qualitative outcomes in 74% of substrates investigated. Model simulations were used to identify the biosynthetic requirements for anaerobic growth of Scheffersomyces stipitis on glucose and the results were validated with published literature. The bottlenecks in Scheffersomyces stipitis metabolic network for xylose uptake and nucleotide cofactor recycling were identified by in silico flux variability analysis. The scope of the model in enhancing the mechanistic understanding of microbial metabolism is demonstrated by identifying a mechanism for mitochondrial respiration and oxidative phosphorylation. Conclusion The genome

Iterative optimization of xylose catabolism in Saccharomyces cerevisiae using combinatorial expression tuning.

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Latimer, Luke N; Dueber, John E

2017-06-01

A common challenge in metabolic engineering is rapidly identifying rate-controlling enzymes in heterologous pathways for subsequent production improvement. We demonstrate a workflow to address this challenge and apply it to improving xylose utilization in Saccharomyces cerevisiae. For eight reactions required for conversion of xylose to ethanol, we screened enzymes for functional expression in S. cerevisiae, followed by a combinatorial expression analysis to achieve pathway flux balancing and identification of limiting enzymatic activities. In the next round of strain engineering, we increased the copy number of these limiting enzymes and again tested the eight-enzyme combinatorial expression library in this new background. This workflow yielded a strain that has a ∼70% increase in biomass yield and ∼240% increase in xylose utilization. Finally, we chromosomally integrated the expression library. This library enriched for strains with multiple integrations of the pathway, which likely were the result of tandem integrations mediated by promoter homology. Biotechnol. Bioeng. 2017;114: 1301-1309. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

Evaluation of UV-C mutagenized Scheffersomyces stipitis strains for ethanol production.

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Geiger, Melanie; Gibbons, Jaimie; West, Thomas; Hughes, Stephen R; Gibbons, William

2012-12-01

We evaluated fermentation capabilities of five strains of Scheffersomyces stipitis (WT-2-1, WT-1-11, 14-2-6, 22-1-1, and 22-1-12) that had been produced by UV-C mutagenesis and selection for improved xylose fermentation to ethanol using an integrated automated robotic work cell. They were incubated under both facultative and anaerobic conditions to evaluate ethanol production on glucose, xylose, cellobiose, and a combination of all three sugars. The medium contained 50 g/L total sugar and 5 g/L yeast extract. The strains performed significantly better under facultative compared with anaerobic conditions. As expected, glucose was the most readily fermented sugar with ~100% fermentation efficiency (FE) under facultative conditions but only 5% to 16% FE anaerobically. Xylose utilization was 20% to 40% FE under facultative conditions but 9% to 25% FE anaerobically. Cellobiose was the least fermented sugar, at 18% to 27% FE facultatively and 8% to 11% anaerobically. Similar trends occurred in the sugar mixture. Under facultative conditions, strain 22-1-12 produced 19.6 g/L ethanol on glucose, but strain 14-2-6 performed best on xylose (4.5 g/L ethanol) and the sugar combination (8.0 g/L ethanol). Ethanol titers from glucose under anaerobic conditions were again highest with strain 22-1-12, but none of the strains produced ethanol from xylose. Future trials will evaluate nutrient addition to boost microaerophilic xylose fermentation.

Lactic acid production from xylose by engineered Saccharomyces cerevisiae without PDC or ADH deletion.

Science.gov (United States)

Turner, Timothy L; Zhang, Guo-Chang; Kim, Soo Rin; Subramaniam, Vijay; Steffen, David; Skory, Christopher D; Jang, Ji Yeon; Yu, Byung Jo; Jin, Yong-Su

2015-10-01

Production of lactic acid from renewable sugars has received growing attention as lactic acid can be used for making renewable and bio-based plastics. However, most prior studies have focused on production of lactic acid from glucose despite that cellulosic hydrolysates contain xylose as well as glucose. Microbial strains capable of fermenting both glucose and xylose into lactic acid are needed for sustainable and economic lactic acid production. In this study, we introduced a lactic acid-producing pathway into an engineered Saccharomyces cerevisiae capable of fermenting xylose. Specifically, ldhA from the fungi Rhizopus oryzae was overexpressed under the control of the PGK1 promoter through integration of the expression cassette in the chromosome. The resulting strain exhibited a high lactate dehydrogenase activity and produced lactic acid from glucose or xylose. Interestingly, we observed that the engineered strain exhibited substrate-dependent product formation. When the engineered yeast was cultured on glucose, the major fermentation product was ethanol while lactic acid was a minor product. In contrast, the engineered yeast produced lactic acid almost exclusively when cultured on xylose under oxygen-limited conditions. The yields of ethanol and lactic acid from glucose were 0.31 g ethanol/g glucose and 0.22 g lactic acid/g glucose, respectively. On xylose, the yields of ethanol and lactic acid were substrates.

Xylose fermentation to ethanol. A review

Energy Technology Data Exchange (ETDEWEB)

McMillan, J D

1993-01-01

The past several years have seen tremendous progress in the understanding of xylose metabolism and in the identification, characterization, and development of strains with improved xylose fermentation characteristics. A survey of the numerous microorganisms capable of directly fermenting xylose to ethanol indicates that wild-type yeast and recombinant bacteria offer the best overall performance in terms of high yield, final ethanol concentration, and volumetric productivity. The best performing bacteria, yeast, and fungi can achieve yields greater than 0.4 g/g and final ethanol concentrations approaching 5%. Productivities remain low for most yeast and particularly for fungi, but volumetric productivities exceeding 1.0 g/L-h have been reported for xylose-fermenting bacteria. In terms of wild-type microorganisms, strains of the yeast Pichia stipitis show the most promise in the short term for direct high-yield fermentation of xylose without byproduct formation. Of the recombinant xylose-fermenting microorganisms developed, recombinant E. coli ATTC 11303 (pLOI297) exhibits the most favorable performance characteristics reported to date.

Moorella stamsii sp. nov., a new anaerobic thermophilic hydrogenogenic carboxydotroph isolated from digester sludge

NARCIS (Netherlands)

Alves, J.I.; Gelder, van A.H.; Alves, M.M.; Sousa, D.Z.; Plugge, C.M.

2013-01-01

A novel anaerobic, thermophilic, carbon monoxide-utilizing bacterium, strain E3-O, was isolated from anaerobic sludge of a municipal solid waste digester. Cells were straight rods, 0.6 to 1µm in diameter and 2 to 3 µm in length, growing as single cells or in pairs. Cells formed round terminal

High ethanol tolerance of the thermophilic anaerobic ethanol producer Thermoanaerobacter BG1L1

DEFF Research Database (Denmark)

Georgieva, Tania I.; Mikkelsen, Marie Just; Ahring, Birgitte Kiær

2007-01-01

The low ethanol tolerance of thermophilic anaerobic bacteria, generally less than 2% (v/v) ethanol, is one of the main limiting factors for their potential use for second generation fuel ethanol production. In this work, the tolerance of thermophilic anaerobic bacterium Thermoanaerobacter BG 1L1...... to exogenously added ethanol was studied in a continuous immobilized reactor system at a growth temperature of 70 degrees C. Ethanol tolerance was evaluated based on inhibition of fermentative performance e.g.. inhibition of substrate conversion. At the highest ethanol concentration tested (8.3% v/v), the strain...... was able to convert 42% of the xylose initially present, indicating that this ethanol concentration is not the upper limit tolerated by the strain. Long-term strain adaptation to high ethanol concentrations (6 - 8.3%) resulted in an improvement of xylose conversion by 25% at an ethanol concentration of 5...

Mutants of Pachysolen tannophilus with Improved Production of Ethanol from d-Xylose †

OpenAIRE

Lee, Hung; James, Allen P.; Zahab, Diana M.; Mahmourides, George; Maleszka, Ryszard; Schneider, Henry

1986-01-01

The conversion of d-xylose to ethanol by the yeast Pachysolen tannophilus is relatively inefficient in batch culture. The inefficiency has been attributed in part to concurrent utilization of ethanol in the presence of appreciable concentrations of d-xylose and to the formation of xylitol and other by-products. To increase the concentration of ethanol accumulated in batch cultures, UV-induced mutants of P. tannophilus were selected on the basis of diminished growth on ethanol. Eleven independ...

Breeding and fermentation characterization of Pachysolen Tannophilus mutant with high ethanol productivity from xylose

International Nuclear Information System (INIS)

Pan Lijun; Chu Kaiqing; Yang Peizhou

2011-01-01

Currently, few strains can utilize xylose to produce ethanol with very low productivity. By the method of mutation breeding to these strains the rate of lignocellulosic utilization could be improved. In this study, the initial Pachysolen tannophilus As 2.1585 was treated by N + ions implantation of 15 keV. The survival curve showed a saddle model. Considering the survival rate and range of positive mutation, the N + ions implantation of 12.5 × 10 14 ions/cm for mutation breeding of Pachysolen tannophilus was selected. A Pachysolen tannophilus mutant mut-54, which had perfect genetic stability of producing ethanol was screened out after continuous 7 passages. The mut-54 had a higher xylose consumption rate, biomass accumulation and ability of ethanol-resistant than the parent strain. Compared with the parent strain, the ethanol concentration fermented by the mut-54 for 72 h increased by 12.74%, which was more suitable for producing ethanol from xylose than the parent strain. (authors)

Creation of a synthetic xylose-inducible promoter for Saccharomyces cerevisiae

Science.gov (United States)

Saccharomyces cerevisiae is currently used to produce ethanol from glucose, but it cannot utilize five-carbon sugars contained in the hemicellulose component of biomass feedstocks. S. cerevisiae strains engineered for xylose fermentation have been made using constitutive promoters to express the req...

Production of 3-hydroxypropionic acid from glucose and xylose by metabolically engineered Saccharomyces cerevisiae

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Kanchana R. Kildegaard

2015-12-01

Full Text Available Biomass, the most abundant carbon source on the planet, may in the future become the primary feedstock for production of fuels and chemicals, replacing fossil feedstocks. This will, however, require development of cell factories that can convert both C6 and C5 sugars present in lignocellulosic biomass into the products of interest. We engineered Saccharomyces cerevisiae for production of 3-hydroxypropionic acid (3HP, a potential building block for acrylates, from glucose and xylose. We introduced the 3HP biosynthetic pathways via malonyl-CoA or β-alanine intermediates into a xylose-consuming yeast. Using controlled fed-batch cultivation, we obtained 7.37±0.17 g 3HP L−1 in 120 hours with an overall yield of 29±1% Cmol 3HP Cmol−1 xylose. This study is the first demonstration of the potential of using S. cerevisiae for production of 3HP from the biomass sugar xylose. Keywords: Metabolic engineering, Biorefineries, 3-hydroxypropionic acid, Saccharomyces cerevisiae, Xylose utilization

Anaerobes in Industrial- and Environmental Biotechnology.

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Hatti-Kaul, Rajni; Mattiasson, Bo

Anaerobic microorganisms present in diverse ecological niches employ alternative strategies for energy conservation in the absence of oxygen which enables them to play a key role in maintaining the global cycles of carbon, nitrogen, and sulfur, and the breakdown of persistent compounds. Thereby they become useful tools in industrial and environmental biotechnology. Although anaerobes have been relatively neglected in comparison to their aerobic counterparts, with increasing knowledge about their diversity and metabolic potential and the development of genetic tools and process technologies to utilize them, we now see a rapid expansion of their applications in the society. This chapter summarizes some of the developments in the use of anaerobes as tools for biomass valorization, in production of energy carriers and chemicals, wastewater treatment, and the strong potential in soil remediation. The ability of several autotrophic anaerobes to reduce carbon dioxide is attracting growing attention as a means for developing a platform for conversion of waste gases to chemicals, materials, and biofuels.

Anaerobic bacteria

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Anaerobic bacteria are bacteria that do not live or grow when oxygen is present. In humans, these bacteria ... Brook I. Diseases caused by non-spore-forming anaerobic bacteria. In: Goldman L, Schafer AI, eds. Goldman-Cecil ...

Development of Efficient Xylose Fermentation in Saccharomyces cerevisiae : Xylose Isomerase as a Key Component

NARCIS (Netherlands)

Van Maris, A.J.A.; Winkler, A.A.; Kuyper, M.; De Laat, W.T.; Van Dijken, J.P.; Pronk, J.T.

2007-01-01

Metabolic engineering of Saccharomyces cerevisiae for ethanol production from d-xylose, an abundant sugar in plant biomass hydrolysates, has been pursued vigorously for the past 15 years. Whereas wild-type S. cerevisiae cannot ferment d-xylose, the ketoisomer d-xylulose can be metabolised slowly.

Breeding of a xylose-fermenting hybrid strain by mating genetically engineered haploid strains derived from industrial Saccharomyces cerevisiae.

Science.gov (United States)

Inoue, Hiroyuki; Hashimoto, Seitaro; Matsushika, Akinori; Watanabe, Seiya; Sawayama, Shigeki

2014-12-01

The industrial Saccharomyces cerevisiae IR-2 is a promising host strain to genetically engineer xylose-utilizing yeasts for ethanol fermentation from lignocellulosic hydrolysates. Two IR-2-based haploid strains were selected based upon the rate of xylulose fermentation, and hybrids were obtained by mating recombinant haploid strains harboring heterogeneous xylose dehydrogenase (XDH) (wild-type NAD(+)-dependent XDH or engineered NADP(+)-dependent XDH, ARSdR), xylose reductase (XR) and xylulose kinase (XK) genes. ARSdR in the hybrids selected for growth rates on yeast extract-peptone-dextrose (YPD) agar and YP-xylose agar plates typically had a higher activity than NAD(+)-dependent XDH. Furthermore, the xylose-fermenting performance of the hybrid strain SE12 with the same level of heterogeneous XDH activity was similar to that of a recombinant strain of IR-2 harboring a single set of genes, XR/ARSdR/XK. These results suggest not only that the recombinant haploid strains retain the appropriate genetic background of IR-2 for ethanol production from xylose but also that ARSdR is preferable for xylose fermentation.

Efficient non-sterilized fermentation of biomass-derived xylose to lactic acid by a thermotolerant Bacillus coagulans NL01.

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Ouyang, Jia; Cai, Cong; Chen, Hai; Jiang, Ting; Zheng, Zhaojuan

2012-12-01

Xylose is the major pentose and the second most abundant sugar in lignocellulosic feedstock. Its efficient utilization is regarded as a technical barrier to the commercial production of bulk chemicals from lignocellulosic biomass. This work aimed at evaluating the lactic acid production from the biomass-derived xylose using non-sterilized fermentation by Bacillus coagulans NL01. A maximum lactic acid concentration of about 75 g/L was achieved from xylose of 100 g/L after 72 h batch fermentation. Acetic acid and levulinic acid were identified as important inhibitors in xylose fermentation, which markedly reduced lactic acid productivity at 15 and 1.0 g/L, respectively. But low concentrations of formic acid (coagulans NL01, the same preference for glucose, xylose, and arabinose was observed and18.2 g/L lactic acid was obtained after 48 h fermentation. These results proved that B. coagulans NL01 was potentially well-suited for producing lactic acid from underutilized xylose-rich prehydrolysates.

Utilization of vinasse effluents from an anaerobic reactor

Energy Technology Data Exchange (ETDEWEB)

Costa, F J.C.B.; Rocha, B B.M.; Viana, C E; Toledo, A C

1986-01-01

An anaerobic reactor was developed to biodigest alcohol distillery wastes. A further post-treatment of the effluent reduced the level of pollution to the point of eventually discharging into streams and rivers. The present work also analyses the use of biodigested vinasse as a source of food for fish. Very high efficiencies were obtained during primary and secondary treatment of vinasse effluent, as demonstrated by the greatly reduced organic load. The utilization of the treated effluent as a source of fish food presents an excellent alternative for the Brazilian alcohol industry. (Refs. 6).

Oxidative production of xylonic acid using xylose in distillation stillage of cellulosic ethanol fermentation broth by Gluconobacter oxydans.

Science.gov (United States)

Zhang, Hongsen; Han, Xushen; Wei, Chengxiang; Bao, Jie

2017-01-01

An oxidative production process of xylonic acid using xylose in distillation stillage of cellulosic ethanol fermentation broth was designed, experimentally investigated, and evaluated. Dry dilute acid pretreated and biodetoxified corn stover was simultaneously saccharified and fermented into 59.80g/L of ethanol (no xylose utilization). 65.39g/L of xylose was obtained in the distillation stillage without any concentrating step after ethanol was distillated. Then the xylose was completely converted into 66.42g/L of xylonic acid by Gluconobacter oxydans. The rigorous Aspen Plus modeling shows that the wastewater generation and energy consumption was significantly reduced comparing to the previous xylonic acid production process using xylose in pretreatment liquid. This study provided a practical process option for xylonic acid production from lignocellulose feedstock with significant reduction of wastewater and energy consumption. Copyright © 2016 Elsevier Ltd. All rights reserved.

Coutilization of D-Glucose, D-Xylose, and L-Arabinose in Saccharomyces cerevisiae by Coexpressing the Metabolic Pathways and Evolutionary Engineering

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Chengqiang Wang

2017-01-01

Full Text Available Efficient and cost-effective fuel ethanol production from lignocellulosic materials requires simultaneous cofermentation of all hydrolyzed sugars, mainly including D-glucose, D-xylose, and L-arabinose. Saccharomyces cerevisiae is a traditional D-glucose fermenting strain and could utilize D-xylose and L-arabinose after introducing the initial metabolic pathways. The efficiency and simultaneous coutilization of the two pentoses and D-glucose for ethanol production in S. cerevisiae still need to be optimized. Previously, we constructed an L-arabinose-utilizing S. cerevisiae BSW3AP. In this study, we further introduced the XI and XR-XDH metabolic pathways of D-xylose into BSW3AP to obtain D-glucose, D-xylose, and L-arabinose cofermenting strain. Benefits of evolutionary engineering: the resulting strain BSW4XA3 displayed a simultaneous coutilization of D-xylose and L-arabinose with similar consumption rates, and the D-glucose metabolic capacity was not decreased. After 120 h of fermentation on mixed D-glucose, D-xylose, and L-arabinose, BSW4XA3 consumed 24% more amounts of pentoses and the ethanol yield of mixed sugars was increased by 30% than that of BSW3AP. The resulting strain BSW4XA3 was a useful chassis for further enhancing the coutilization efficiency of mixed sugars for bioethanol production.

Lactic acid production from xylose by engineered Saccharomyces cerevisiae without PDC or ADH deletion

Science.gov (United States)

Production of lactic acid from renewable sugars has received growing attention as lactic acid can be used for making renewable and bio-based plastics. However, most prior studies have focused on production of lactic acid from glucose despite cellulosic hydrolysates contain xylose as well as glucose....

Xylose Fermentation by Saccharomyces cerevisiae: Challenges and Prospects

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Danuza Nogueira Moysés

2016-02-01

Full Text Available Many years have passed since the first genetically modified Saccharomyces cerevisiae strains capable of fermenting xylose were obtained with the promise of an environmentally sustainable solution for the conversion of the abundant lignocellulosic biomass to ethanol. Several challenges emerged from these first experiences, most of them related to solving redox imbalances, discovering new pathways for xylose utilization, modulation of the expression of genes of the non-oxidative pentose phosphate pathway, and reduction of xylitol formation. Strategies on evolutionary engineering were used to improve fermentation kinetics, but the resulting strains were still far from industrial application. Lignocellulosic hydrolysates proved to have different inhibitors derived from lignin and sugar degradation, along with significant amounts of acetic acid, intrinsically related with biomass deconstruction. This, associated with pH, temperature, high ethanol, and other stress fluctuations presented on large scale fermentations led the search for yeasts with more robust backgrounds, like industrial strains, as engineering targets. Some promising yeasts were obtained both from studies of stress tolerance genes and adaptation on hydrolysates. Since fermentation times on mixed-substrate hydrolysates were still not cost-effective, the more selective search for new or engineered sugar transporters for xylose are still the focus of many recent studies. These challenges, as well as under-appreciated process strategies, will be discussed in this review.

Enhanced production of extracellular inulinase by the yeast Kluyveromyces marxianus in xylose catabolic state.

Science.gov (United States)

Hoshida, Hisashi; Kidera, Kenta; Takishita, Ryuta; Fujioka, Nobuhisa; Fukagawa, Taiki; Akada, Rinji

2018-06-01

The production of extracellular proteins by the thermotolerant yeast Kluyveromyces marxianus, which utilizes various sugars, was investigated using media containing sugars such as glucose, galactose, and xylose. SDS-PAGE analysis of culture supernatants revealed abundant production of an extracellular protein when cells were grown in xylose medium. The N-terminal sequence of the extracellular protein was identical to a part of the inulinase encoded by INU1 in the genome. Inulinase is an enzyme hydrolyzing β-2,1-fructosyl bond in inulin and sucrose and is not required for xylose assimilation. Disruption of INU1 in the strain DMKU 3-1042 lost the production of the extracellular protein and resulted in growth defect in sucrose and inulin media, indicating that the extracellular protein was inulinase (sucrase). In addition, six K. marxianus strains among the 16 strains that were analyzed produced more inulinase in xylose medium than in glucose medium. However, expression analysis indicated that the INU1 promoter activity was lower in the xylose medium than in the glucose medium, suggesting that enhanced production of inulinase is controlled in a post-transcriptional manner. The production of inulinase was also higher in cultures with more agitation, suggesting that oxygen supply affects the production of inulinase. Taken together, these results suggest that both xylose and oxygen supply shift cellular metabolism to enhance the production of extracellular inulinase. Copyright © 2018 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Metabolic control analysis of xylose catabolism in Aspergillus

NARCIS (Netherlands)

Prathumpai, W.; Gabelgaard, J.B.; Wanchanthuek, P.; Vondervoort, van de P.J.I.; Groot, de M.J.L.; McIntyre, M.; Nielsen, J.

2003-01-01

A kinetic model for xylose catabolism in Aspergillus is proposed. From a thermodynamic analysis it was found that the intermediate xylitol will accumulate during xylose catabolism. Use of the kinetic model allowed metabolic control analysis (MCA) of the xylose catabolic pathway to be carried out,

Succinic acid production from xylose mother liquor by recombinant Escherichia coli strain.

Science.gov (United States)

Wang, Honghui; Pan, Jiachuan; Wang, Jing; Wang, Nan; Zhang, Jie; Li, Qiang; Wang, Dan; Zhou, Xiaohua

2014-11-02

Succinic acid (1,4-butanedioic acid) is identified as one of important building-block chemicals. Xylose mother liquor is an abundant industrial residue in xylitol biorefining industry. In this study, xylose mother liquor was utilized to produce succinic acid by recombinant Escherichia coli strain SD121, and the response surface methodology was used to optimize the fermentation media. The optimal conditions of succinic acid fermentation were as follows: 82.62 g L -1 total initial sugars, 42.27 g L -1 MgCO 3 and 17.84 g L -1 yeast extract. The maximum production of succinic acid was 52.09 ± 0.21 g L -1 after 84 h with a yield of 0.63 ± 0.03 g g -1 total sugar, approaching the predicted value (53.18 g L -1 ). It was 1.78-fold of the production of that obtained with the basic medium. This was the first report on succinic acid production from xylose mother liquor by recombinant E. coli strains with media optimization using response surface methodology. This work suggested that the xylose mother liquor could be an alternative substrate for the economical production of succinic acid by recombinant E. coli strains.

Metabolic Engineering for Substrate Co-utilization

Science.gov (United States)

Gawand, Pratish

Production of biofuels and bio-based chemicals is being increasingly pursued by chemical industry to reduce its dependence on petroleum. Lignocellulosic biomass (LCB) is an abundant source of sugars that can be used for producing biofuels and bio-based chemicals using fermentation. Hydrolysis of LCB results in a mixture of sugars mainly composed of glucose and xylose. Fermentation of such a sugar mixture presents multiple technical challenges at industrial scale. Most industrial microorganisms utilize sugars in a sequential manner due to the regulatory phenomenon of carbon catabolite repression (CCR). Due to sequential utilization of sugars, the LCB-based fermentation processes suffer low productivities and complicated operation. Performance of fermentation processes can be improved by metabolic engineering of microorganisms to obtain superior characteristics such as high product yield. With increased computational power and availability of complete genomes of microorganisms, use of model-based metabolic engineering is now a common practice. The problem of sequential sugar utilization, however, is a regulatory problem, and metabolic models have never been used to solve such regulatory problems. The focus of this thesis is to use model-guided metabolic engineering to construct industrial strains capable of co-utilizing sugars. First, we develop a novel bilevel optimization algorithm SimUp, that uses metabolic models to identify reaction deletion strategies to force co-utilization of two sugars. We then use SimUp to identify reaction deletion strategies to force glucose-xylose co-utilization in Escherichia coli. To validate SimUp predictions, we construct three mutants with multiple gene knockouts and test them for glucose-xylose utilization characteristics. Two mutants, designated as LMSE2 and LMSE5, are shown to co-utilize glucose and xylose in agreement with SimUp predictions. To understand the molecular mechanism involved in glucose-xylose co-utilization of the

Anaerobic Pseudomonas aeruginosa and other obligately anaerobic bacterial biofilms growing in the thick airway mucus of chronically infected cystic fibrosis patients: an emerging paradigm or "Old Hat"?

Science.gov (United States)

Su, Shengchang; Hassett, Daniel J

2012-09-01

The cystic fibrosis (CF) airway mucus is an ideal niche in which many bacteria can develop antibiotic- and phagocyte-resistance in unique structures known as "mode II biofilms" where bacteria are embedded within the mucus, yet unattached to airway epithelial cells. Pseudomonas aeruginosa is the dominant CF pathogen, yet herein the authors provide burgeoning evidence that obligate anaerobic bacteria (e.g., Prevotella) actually thrive within the CF mucus, a paradigmatic shift that chronic CF is an "aerobic" disease. Interestingly, CF organisms repress virulence factor production (e.g., P. aeruginosa) while others (e.g., S. aureus) increase them under anaerobic conditions. The authors shed additional light on (i) the anoxic nature of the CF airway mucus, (ii) the relative commonality of anaerobic bacteria isolated from CF sputum, (iii) virulence factor production and cross-talk between obligate anaerobes and P. aeruginosa relative to disease progression/remission, (iv) the role of mucoidy in CF, and (v) the role of nitrosative stress in activation of bacteriophage and pyocins within biofilms. The authors conclude with insight as to how we might treat some CF bacteria during mode II biofilm infections that utilizes a metabolite of bacterial anaerobic respiration and an aerobic oxidation product of airway-generated NO, acidified NO(2)(-).

Integrated biogas upgrading and hydrogen utilization in an anaerobic reactor containing enriched hydrogenotrophic methanogenic culture

DEFF Research Database (Denmark)

Luo, Gang; Angelidaki, Irini

2012-01-01

Biogas produced by anaerobic digestion, is mainly used in a gas motor for heat and electricity production. However, after removal of CO2, biogas can be upgraded to natural gas quality, giving more utilization possibilities, such as utilization as autogas, or distant utilization by using...... the existing natural gas grid. The current study presents a new biological method for biogas upgrading in a separate biogas reactor, containing enriched hydrogenotrophic methanogens and fed with biogas and hydrogen. Both mesophilic- and thermophilic anaerobic cultures were enriched to convert CO2 to CH4...... by addition of H2. Enrichment at thermophilic temperature (55°C) resulted in CO2 and H2 bioconversion rate of 320 mL CH4/(gVSS h), which was more than 60% higher than that under mesophilic temperature (37°C). Different dominant species were found at mesophilic- and thermophilic-enriched cultures, as revealed...

Application of dynamic membranes in anaerobic membranes in anaerobic membrane bioreactor systems

NARCIS (Netherlands)

Erşahin, M.E.

2015-01-01

Anaerobic membrane bioreactors (AnMBRs) physically ensure biomass retention by the application of a membrane filtration process. With growing application experiences from aerobic membrane bioreactors (MBRs), the combination of membrane and anaerobic processes has received much attention and become

Characterization of xylose reductase from Candida tropicalis ...

African Journals Online (AJOL)

USER

2010-08-02

Aug 2, 2010 ... production are the possibility of using industrial side- streams as raw ... xylitol production,. D-xylose assimilation in microorganism involves xylose ..... natural biopolymer extracted from brown alga, and in the presence of ...

In Search of functionality-diversity relationships in anaerobic mixed culture fermentations

International Nuclear Information System (INIS)

Kleerebezem, R.; Temudo, M.; Muyzer Van Loosdrecht, M. C. M.

2009-01-01

Based on the work described in this paper we will postulate that in environmental ecosystems with a weak selective pressure no clear relationship exists between the ecosystem functionality and the microbial diversity and microbial composition. In the past years we have been investigating the anaerobic fermentation of glucose, xylose, and glycerol, and mixtures of these substrates in continuously stirred tank reactors (CSTR) inoculated with an activated sludge characterized by a very rich microbial diversity. (Author)

Biotechnological Utilization with a Focus on Anaerobic Treatment of Cheese Whey: Current Status and Prospects

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Aspasia A. Chatzipaschali

2012-09-01

Full Text Available Cheese whey utilization is of major concern nowadays. Its high organic matter content, in combination with the high volumes produced and limited treatment options make cheese whey a serious environmental problem. However, the potential production of biogas (methane, hydrogen or other marketable products with a simultaneous high COD reduction through appropriate treatment proves that cheese whey must be considered as an energy resource rather than a pollutant. The presence of biodegradable components in the cheese whey coupled with the advantages of anaerobic digestion processes over other treatment methods makes anaerobic digestion an attractive and suitable treatment option. This paper intends to review the most representative applications of anaerobic treatment of cheese whey currently being exploited and under research. Moreover, an effort has been made to categorize the common characteristics of the various research efforts and find a comparative basis, as far as their results are concerned. In addition, a number of dairy industries already using such anaerobic digestion systems are presented.

Proteomic analysis of the secretory response of Aspergillus niger to D-maltose and D-xylose.

Directory of Open Access Journals (Sweden)

José Miguel P Ferreira de Oliveira

Full Text Available Fungi utilize polysaccharide substrates through extracellular digestion catalyzed by secreted enzymes. Thus far, protein secretion by the filamentous fungus Aspergillus niger has mainly been studied at the level of individual proteins and by genome and transcriptome analyses. To extend these studies, a complementary proteomics approach was applied with the aim to investigate the changes in secretome and microsomal protein composition resulting from a shift to a high level secretion condition. During growth of A. niger on D-sorbitol, small amounts of D-maltose or D-xylose were used as inducers of the extracellular amylolytic and xylanolytic enzymes. Upon induction, protein compositions in the extracellular broth as well as in enriched secretory organelle (microsomal fractions were analyzed using a shotgun proteomics approach. In total 102 secreted proteins and 1,126 microsomal proteins were identified in this study. Induction by D-maltose or D-xylose resulted in the increase in specific extracellular enzymes, such as glucoamylase A on D-maltose and β-xylosidase D on D-xylose, as well as of microsomal proteins. This reflects the differential expression of selected genes coding for dedicated extracellular enzymes. As expected, the addition of extra D-sorbitol had no effect on the expression of carbohydrate-active enzymes, compared to addition of D-xylose or D-maltose. Furthermore, D-maltose induction caused an increase in microsomal proteins related to translation (e.g., Rpl15 and vesicular transport (e.g., the endosomal-cargo receptor Erv14. Millimolar amounts of the inducers D-maltose and D-xylose are sufficient to cause a direct response in specific protein expression levels. Also, after induction by D-maltose or D-xylose, the induced enzymes were found in microsomes and extracellular. In agreement with our previous findings for D-xylose induction, D-maltose induction leads to recruitment of proteins involved in proteasome-mediated degradation.

Proteomic analysis of the secretory response of Aspergillus niger to D-maltose and D-xylose.

Science.gov (United States)

de Oliveira, José Miguel P Ferreira; van Passel, Mark W J; Schaap, Peter J; de Graaff, Leo H

2011-01-01

Fungi utilize polysaccharide substrates through extracellular digestion catalyzed by secreted enzymes. Thus far, protein secretion by the filamentous fungus Aspergillus niger has mainly been studied at the level of individual proteins and by genome and transcriptome analyses. To extend these studies, a complementary proteomics approach was applied with the aim to investigate the changes in secretome and microsomal protein composition resulting from a shift to a high level secretion condition. During growth of A. niger on D-sorbitol, small amounts of D-maltose or D-xylose were used as inducers of the extracellular amylolytic and xylanolytic enzymes. Upon induction, protein compositions in the extracellular broth as well as in enriched secretory organelle (microsomal) fractions were analyzed using a shotgun proteomics approach. In total 102 secreted proteins and 1,126 microsomal proteins were identified in this study. Induction by D-maltose or D-xylose resulted in the increase in specific extracellular enzymes, such as glucoamylase A on D-maltose and β-xylosidase D on D-xylose, as well as of microsomal proteins. This reflects the differential expression of selected genes coding for dedicated extracellular enzymes. As expected, the addition of extra D-sorbitol had no effect on the expression of carbohydrate-active enzymes, compared to addition of D-xylose or D-maltose. Furthermore, D-maltose induction caused an increase in microsomal proteins related to translation (e.g., Rpl15) and vesicular transport (e.g., the endosomal-cargo receptor Erv14). Millimolar amounts of the inducers D-maltose and D-xylose are sufficient to cause a direct response in specific protein expression levels. Also, after induction by D-maltose or D-xylose, the induced enzymes were found in microsomes and extracellular. In agreement with our previous findings for D-xylose induction, D-maltose induction leads to recruitment of proteins involved in proteasome-mediated degradation.

[Utility of MALDI-TOF MS for the identification of anaerobic bacteria].

Science.gov (United States)

Zárate, Mariela S; Romano, Vanesa; Nievas, Jimena; Smayevsky, Jorgelina

2014-01-01

The analysis by MALDI-TOF MS (Matrix-assited laser desorption/ionization time-of-flight mass spectrometry) has become a reference method for the identification of microorganisms in Clinical Microbiology. However, data on some groups of microorganisms are still controversial. The aim of this study is to determine the utility of MALDI-TOF MS for the identification of clinical isolates of anaerobic bacteria. One-hundred and six anaerobic bacteria isolates were analyzed by MALDI-TOF MS and by conventional biochemical tests. In those cases where identification by conventional methodology was not applicable or in the face of discordance between sequencing methodologies, 16 S rRNA gene sequence analysis was performed. The conventional method and MALDI-TOF MS agreed at genus and species level by 95.3 %. Concordance in gram-negative bacilli was 91.4% and 100% among gram-positive bacilli; there was also concordance both in the 8 isolates studied in gram-positive cocci and in the single gram-negative cocci included. The data obtained in this study demonstrate that MALDI-TOF MS offers the possibility of adequate identification of anaerobic bacteria. Copyright © 2014 Asociación Colombiana de Psiquiatría. Publicado por Elsevier España. All rights reserved.

Co-fermentation of glucose, xylose and/or cellobiose by yeast

Science.gov (United States)

Jeffries, Thomas W.; Willis, Laura B.; Long, Tanya M.; Su, Yi-Kai

2013-09-10

Provided herein are methods of using yeast cells to produce ethanol by contacting a mixture comprising xylose with a Spathaspora yeast cell under conditions suitable to allow the yeast to ferment at least a portion of the xylose to ethanol. The methods allow for efficient ethanol production from hydrolysates derived from lignocellulosic material and sugar mixtures including at least xylose and glucose or xylose, glucose and cellobiose.

Nutritional implications of D-xylose in pigs

NARCIS (Netherlands)

Schutte, J.B.; Jong, J.de; Polziehn, R.; Verstegen, M.W.A.

1991-01-01

Hemicellulose consists primarily of pentose sugars, joined together in a polysaccharide chain with D-xylose as the most abundant component. Ileal digestibility and urinary excretion of D-xylose and associated effects of this pentose sugar on ileal and faecal digestibility of dry matter (DM), organic

Fermentation of Xylose Causes Inefficient Metabolic State Due to Carbon/Energy Starvation and Reduced Glycolytic Flux in Recombinant Industrial Saccharomyces cerevisiae

Science.gov (United States)

Matsushika, Akinori; Nagashima, Atsushi; Goshima, Tetsuya; Hoshino, Tamotsu

2013-01-01

In the present study, comprehensive, quantitative metabolome analysis was carried out on the recombinant glucose/xylose-cofermenting S. cerevisiae strain MA-R4 during fermentation with different carbon sources, including glucose, xylose, or glucose/xylose mixtures. Capillary electrophoresis time-of-flight mass spectrometry was used to determine the intracellular pools of metabolites from the central carbon pathways, energy metabolism pathways, and the levels of twenty amino acids. When xylose instead of glucose was metabolized by MA-R4, glycolytic metabolites including 3- phosphoglycerate, 2- phosphoglycerate, phosphoenolpyruvate, and pyruvate were dramatically reduced, while conversely, most pentose phosphate pathway metabolites such as sedoheptulose 7- phosphate and ribulose 5-phosphate were greatly increased. These results suggest that the low metabolic activity of glycolysis and the pool of pentose phosphate pathway intermediates are potential limiting factors in xylose utilization. It was further demonstrated that during xylose fermentation, about half of the twenty amino acids declined, and the adenylate/guanylate energy charge was impacted due to markedly decreased adenosine triphosphate/adenosine monophosphate and guanosine triphosphate/guanosine monophosphate ratios, implying that the fermentation of xylose leads to an inefficient metabolic state where the biosynthetic capabilities and energy balance are severely impaired. In addition, fermentation with xylose alone drastically increased the level of citrate in the tricarboxylic acid cycle and increased the aromatic amino acids tryptophan and tyrosine, strongly supporting the view that carbon starvation was induced. Interestingly, fermentation with xylose alone also increased the synthesis of the polyamine spermidine and its precursor S-adenosylmethionine. Thus, differences in carbon substrates, including glucose and xylose in the fermentation medium, strongly influenced the dynamic metabolism of MA-R4

Pilot-scale steam explosion for xylose production from oil palm empty fruit bunches and the use of xylose for ethanol production.

Science.gov (United States)

Duangwang, Sairudee; Ruengpeerakul, Taweesak; Cheirsilp, Benjamas; Yamsaengsung, Ram; Sangwichien, Chayanoot

2016-03-01

Pilot-scale steam explosion equipments were designed and constructed, to experimentally solubilize xylose from oil palm empty fruit bunches (OPEFB) and also to enhance an enzyme accessibility of the residual cellulose pulp. The OPEFB was chemically pretreated prior to steam explosion at saturated steam (SS) and superheated steam (SHS) conditions. The acid pretreated OPEFB gave the highest xylose recovery of 87.58 ± 0.21 g/kg dried OPEFB in the liquid fraction after explosion at SHS condition. These conditions also gave the residual cellulose pulp with high enzymatic accessibility of 73.54 ± 0.41%, which is approximately threefold that of untreated OPEFB. This study has shown that the acid pretreatment prior to SHS explosion is an effective method to enhance both xylose extraction and enzyme accessibility of the exploded OPEFB. Moreover, the xylose solution obtained in this manner could directly be fermented by Candida shehatae TISTR 5843 giving high ethanol yield of 0.30 ± 0.08 g/g xylose. Copyright © 2015 Elsevier Ltd. All rights reserved.

Thermochemistry of α-D-xylose(cr)

International Nuclear Information System (INIS)

Ribeiro da Silva, Manuel A.V.; Ribeiro da Silva, Maria D.M.C.; Lobo Ferreira, Ana I.M.C.; Shi, Quan; Woodfield, Brian F.; Goldberg, Robert N.

2013-01-01

Highlights: ► Well-characterized material. ► Oxygen bomb calorimetry. ► Heat capacities obtained by using a Physical Property Measurement System. ► Thermochemical Network Calculations. ► Accurate thermodynamic property values of a key biochemical substance. -- Abstract: The thermochemistry of α-D-xylose(cr) was studied by means of oxygen bomb calorimetry and a Physical Property Measurement System (PPMS) in zero magnetic field. The sample of α-D-xylose(cr) used in this study was one well-characterized by HPLC, Karl Fischer analysis, NMR, and by carbon dioxide analysis. The standard molar enthalpy of combustion was found to be Δ c H m o = −(2342.2 ± 0.8) kJ·mol −1 at T = 298.15 K and at the standard pressure p° = 0.1 MPa. The standard molar heat capacity for α-D-xylose(cr) was measured with the PPMS over the temperature range 1.9001 ⩽ T/K ⩽ 303.66. At T = 298.15 K, C p,m o = (178.1 ± 1.8) J·K −1 ·mol −1 . The values of C p,m o were fit as a function of T by using theoretical and empirical models for appropriate temperature ranges. The results of these fits were used to calculate values of C p,m o , the entropy increment Δ 0 T S m o , Δ 0 T H m o , and Φ m o =(Δ 0 T S m o -Δ 0 T H m o /T) from T = 0.5 K to T = 300 K. Derived quantities for α-D-xylose(cr) are the standard molar enthalpy of formation Δ f H m o = −(1054.5 ± 1.1) kJ·mol −1 , the third law standard molar entropy S m o = (175.3 ± 1.9) J·K −1 ·mol −1 , and the standard molar Gibbs energy of formation Δ f G m o = −(750.5 ± 1.0) kJ·mol −1 . A comparison of values of Δ c H m o and S m o for the five-carbon aldoses demonstrated a striking similarity in the values of these respective properties for α-D-xylose(cr), D-ribose(cr), and D-arabinose(cr). Thermochemical network calculations were performed that led to values of the standard formation properties at T = 298.15 K for a variety of biochemical substances: D-xylose(aq), D-xylose − (aq), D-xylose 2�

Purification and characterization of the d-xylose isomerase gene from Escherichia coli

Energy Technology Data Exchange (ETDEWEB)

Ho, N W.Y.; Rosenfeld, S; Stevis, P; Tsao, G T

1983-11-01

A DNA fragment containing both the Escherichia coli D-xylose isomerase (D-xylose ketol-isomerase, EC 5.3.1.5) gene and the D-xylulokinase (ATP: D-xylulose 5-phosphotransferase, EC 2.7.1.17) gene has been cloned on an E. coli plasmid. The D-xylose isomerase gene was separated from the D-xylulokinase gene by the construction of a new deletion plasmid, pLX7. The D-xylose isomerase gene cloned on pLX7 was found still to be an intact gene. The precise location of the D-xylose isomerase gene on the plasmid pLX7 was further determined by the construction of two more plasmids, pLX8 and pLX9. This is believed to be the first D-xylose isomerase gene that has been isolated and extensively purified from any organism. D-Xylose isomerase, the enzyme product of the D-xylose isomerase gene, is responsible for the conversion of D-xylose to D-xylulose, as well as D-glucose to D-fructose. It is widely believed that yeast cannot ferment D-xylose to ethanol primarily because of the lack of D-xylose isomerase in yeast. D-Xylose isomerase (also known as D-glucose isomerase) is also used for the commercial production of high-fructose syrups. The purification of the D-xylose isomerase gene may lead to the following industrial applications: (1) cloning and expression of the gene in yeast to make the latter organism capable of directly fermenting D-xylose to ethanol, and (2) cloning of the gene on a high-copy-number plasmid in a proper host to overproduce the enzyme, which should have a profound impact on the high-fructose syrup technology. 14 references.

Changing flux of xylose metabolites by altering expression of xylose reductase and xylitol dehydrogenase in recombinant Saccharomyces cerevisiae

Science.gov (United States)

Yong-Su Jin; Thomas W. Jeffries

2003-01-01

We changed the fluxes of xylose metabolites in recombinant Saccharomyces cerevisiae by manipulating expression of Pichia stipitis genes(XYL1 and XYL2) coding for xylose reductase (XR) and xylitol dehydrogenase (XDH), respectively. XYL1 copy number was kept constant by integrating it into the chromosome. Copy numbers of XYL2 were varied either by integrating XYL2 into...

Cell growth and hydrogen production on the mixture of xylose and glucose using a novel strain of Clostridium sp. HR-1 isolated from cow dung compost

Energy Technology Data Exchange (ETDEWEB)

Xu, Ji-Fei; Ren, Nan-Qi; Wang, Ai-Jie; Qiu, Jie; Zhao, Qing-Liang; Feng, Yu-Jie; Liu, Bing-Feng [State Key Laboratory of Urban Water Resource and Environment (SKLUWRE), School of Municipal and Environmental Engineering, Harbin Institute of Technology, Harbin 150090 (China)

2010-12-15

A novel mesophilic hydrogen-producing bacterium was isolated from cow dung compost and designated as Clostridium sp. HR-1 by 16S rRNA gene sequence. The optimum condition for hydrogen production by strain HR-1 was pH of 6.5, temperature of 37 C and yeast extract as nitrogen sources. The strain HR-1 has the ability to utilize kinds of hexose and pentose as carbon sources for growth and H{sub 2} production. Cell growth and hydrogen productivity were investigated for batch fermentation on media containing different ratios of xylose and glucose. Glucose was the preferred substrate in the glucose and xylose mixtures. The high glucose fraction had higher cell biomass production rate. The rate of glucose consumption was higher than xylose consumption, and remained essentially constant independent of xylose content of the mixture. The rate of xylose utilization was decreased with increasing of the glucose fraction. The average H{sub 2} yield and specific H{sub 2} production rates with xylose and glucose are 1.63 mol-H{sub 2}/mol xylose and 11.14-H{sub 2} mmol/h g-cdw, and 2.02 mol-H{sub 2}/mol-glucose and 9.37 mmol-H{sub 2}/h g-cdw, respectively. Using the same initial substrate concentration, the maximum average H{sub 2} yield and specific H{sub 2} production rates with the mixtures of 9 g/l xylose and 3 g/l glucose was 2.01 mol-H{sub 2}/mol-mixed sugar and 12.56 mmol-H{sub 2}/h g-cdw, respectively. During the fermentation, the main soluble microbial products were ethanol and acetate which showed trends with the different ratios of xylose and glucose. (author)

Stoichiometric network constraints on xylose metabolism by recombinant Saccharomyces cerevisiae

Science.gov (United States)

Yong-Su Jin; Thomas W. Jeffries

2004-01-01

Metabolic pathway engineering is constrained by the thermodynamic and stoichiometric feasibility of enzymatic activities of introduced genes. Engineering of xylose metabolism in Saccharomyces cerevisiae has focused on introducing genes for the initial xylose assimilation steps from Pichia stipitis, a xylose-fermenting yeast, into S. cerevisiae, a yeast raditionally...

Improved ethanol production from xylose in the presence of acetic acid by the overexpression of the HAA1 gene in Saccharomyces cerevisiae.

Science.gov (United States)

Sakihama, Yuri; Hasunuma, Tomohisa; Kondo, Akihiko

2015-03-01

The hydrolysis of lignocellulosic biomass liberates sugars, primarily glucose and xylose, which are subsequently converted to ethanol by microbial fermentation. The rapid and efficient fermentation of xylose by recombinant Saccharomyces cerevisiae strains is limited by weak acids generated during biomass pretreatment processes. In particular, acetic acid negatively affects cell growth, xylose fermentation rate, and ethanol production. The ability of S. cerevisiae to efficiently utilize xylose in the presence of acetic acid is an essential requirement for the cost-effective production of ethanol from lignocellulosic hydrolysates. Here, an acetic acid-responsive transcriptional activator, HAA1, was overexpressed in a recombinant xylose-fermenting S. cerevisiae strain to yield BY4741X/HAA1. This strain exhibited improved cell growth and ethanol production from xylose under aerobic and oxygen limited conditions, respectively, in the presence of acetic acid. The HAA1p regulon enhanced transcript levels in BY4741X/HAA1. The disruption of PHO13, a p-nitrophenylphosphatase gene, in BY4741X/HAA1 led to further improvement in both yeast growth and the ability to ferment xylose, indicating that HAA1 overexpression and PHO13 deletion act by different mechanisms to enhance ethanol production. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Enhanced L-lactic acid production from biomass-derived xylose by a mutant Bacillus coagulans.

Science.gov (United States)

Zheng, Zhaojuan; Cai, Cong; Jiang, Ting; Zhao, Mingyue; Ouyang, Jia

2014-08-01

Xylose effective utilization is crucial for production of bulk chemicals from low-cost lignocellulosic substrates. In this study, an efficient L-lactate production process from xylose by a mutant Bacillus coagulans NL-CC-17 was demonstrated. The nutritional requirements for L-lactate production by B. coagulans NL-CC-17 were optimized statistically in shake flask fermentations. Corn steep liquor powder and yeast exact were identified as the most significant factors by the two-level Plackett-Burman design. Steepest ascent experiments were applied to approach the optimal region of the two factors, and a central composite design was employed to determine their optimal levels. The optimal medium was used to perform batch fermentation in a 3-l bioreactor. A maximum of 90.29 g l(-1)  L-lactic acid was obtained from 100 g l(-1) xylose in 120 h. When using corn stove prehydrolysates as substrates, 23.49 g l(-1)  L-lactic acid was obtained in 36 h and the yield was 83.09 %.

Improvement of Xylose Fermentation Ability under Heat and Acid Co-Stress in Saccharomyces cerevisiae Using Genome Shuffling Technique

Directory of Open Access Journals (Sweden)

Kentaro Inokuma

2017-12-01

Full Text Available Xylose-assimilating yeasts with tolerance to both fermentation inhibitors (such as weak organic acids and high temperature are required for cost-effective simultaneous saccharification and cofermentation (SSCF of lignocellulosic materials. Here, we demonstrate the construction of a novel xylose-utilizing Saccharomyces cerevisiae strain with improved fermentation ability under heat and acid co-stress using the drug resistance marker-aided genome shuffling technique. The mutagenized genome pools derived from xylose-utilizing diploid yeasts with thermotolerance or acid tolerance were shuffled by sporulation and mating. The shuffled strains were then subjected to screening under co-stress conditions of heat and acids, and the hybrid strain Hyb-8 was isolated. The hybrid strain displayed enhanced xylose fermentation ability in comparison to both parental strains under co-stress conditions of heat and acids. Hyb-8 consumed 33.1 ± 0.6 g/L xylose and produced 11.1 ± 0.4 g/L ethanol after 72 h of fermentation at 38°C with 20 mM acetic acid and 15 mM formic acid. We also performed transcriptomic analysis of the hybrid strain and its parental strains to screen for key genes for multiple stress tolerances. We found that 13 genes, including 5 associated with cellular transition metal ion homeostasis, were significantly upregulated in Hyb-8 compared to levels in both parental strains under co-stress conditions. The hybrid strain Hyb-8 has strong potential for cost-effective SSCF of lignocellulosic materials. Moreover, the transcriptome data gathered in this study will be useful for understanding the mechanisms of multiple tolerance to high temperature and acids in yeast and facilitate the development of robust yeast strains for SSCF.

Saccharomyces cerevisiae engineered for xylose metabolism requires gluconeogenesis and the oxidative branch of the pentose phosphate pathway for aerobic xylose assimilation

Science.gov (United States)

Saccharomyces strains engineered to ferment xylose using Scheffersomyces stipitis xylose reductase (XR) and xylitol dehydrogenase (XDH) genes appear to be limited by metabolic imbalances due to differing cofactor specificities of XR and XDH. The S. stipitis XR, which uses nicotinamide adenine dinucl...

The effect of CreA in glucose and xylose catabolism in Aspergillus nidulans

DEFF Research Database (Denmark)

Prathumpai, Wai; Mcintyre, Mhairi; Nielsen, Jens

2004-01-01

The catabolism of glucose and xylose was studied in a wild type and creA deleted (carbon catabolite de-repressed) strain of Aspergillus nidulans. Both strains were cultivated in bioreactors with either glucose or xylose as the sole carbon source, or in the presence of both sugars. In the cultivat......The catabolism of glucose and xylose was studied in a wild type and creA deleted (carbon catabolite de-repressed) strain of Aspergillus nidulans. Both strains were cultivated in bioreactors with either glucose or xylose as the sole carbon source, or in the presence of both sugars...... on the sugar mixture, glucose repression of xylose utilisation was observed; with xylose utilisation occurring only after glucose was depleted. This phenomenon was not seen in the creA deleted strain, where glucose and xylose were catabolised simultaneously. Measurement of key metabolites and the activities...... of key enzymes in the xylose utilisation pathway revealed that xylose metabolism was occurring in the creA deleted strain, even at high glucose concentrations. Conversely, in the wild type strain, activities of the key enzymes for xylose metabolism increased only when the effects of glucose repression...

Metabolic characterization and transformation of the non-dairy Lactococcus lactis strain KF147, for production of ethanol from xylose

DEFF Research Database (Denmark)

Petersen, Kia Vest; Liu, Jianming; Chen, Jun

2017-01-01

producing ethanol as the sole fermentation product with a high yield corresponding to 83% of the theoretical maximum. The results clearly indicate the great potential of using the more metabolically diverse non-dairy L. lactis strains for bio-production based on xylose containing feedstocks.......The non-dairy lactic acid bacterium Lactococcus lactis KF147 can utilize xylose as the sole energy source. To assess whether KF147 could serve as a platform organism for converting second generation sugars into useful chemicals, we characterized growth and product formation for KF147 when grown...... the arcA gene encoding the arginine deiminase. The fermentation product profile suggested two routes for xylose degradation, the phosphoketolase pathway and the pentose phosphate pathway. Inactivation of the phosphoketolase pathway redirected the entire flux through the pentose phosphate pathway whereas...

Comparative evaluation of anoxomat and conventional anaerobic GasPak jar systems for the isolation of anaerobic bacteria.

Science.gov (United States)

Shahin, May; Jamal, Wafaa; Verghese, Tina; Rotimi, V O

2003-01-01

To evaluate the performance of the Anoxomat, in comparison with the conventional anaerobic GasPak jar system, for the isolation of obligate anaerobes. Anoxomat, model WS800, and anaerobic GasPak jar system (Oxoid) were evaluated. Anoxomat system utilized a gas mixture of 80% N(2), 10% CO(2) and 10% H(2), while the GasPak used a gas mixture of 90% H(2) and 10% CO(2). An anaerobic indicator within the jars monitored anaerobiosis. A total of 227 obligate anaerobic bacteria comprising 116 stock strains, 5 ATCC reference strains and 106 fresh strains, representing different genera, were investigated for growth on anaerobic agar plates and scored for density, colony sizes, susceptibility zones of antibiotic inhibition and the speed of anaerobiosis (reducing the indicator). The results demonstrate that the growth of anaerobic bacteria is faster inside the Anoxomat jar than in the anaerobic GasPak jar system. Of the 227 strains tested, the colonies of 152 (67%) were larger (by size range of 0.2-2.4 mm) in the Anoxomat at 48 h than in the GasPak jar compared with only 21% (range 0.1-0.3 mm) that were larger in the GasPak than in the Anoxomat. The remaining 12% were equal in their sizes. There was no measurable difference in the colony sizes of the reference strains. The Porphyromonas asaccharolytica strains failed to grow within the GasPak system but grew inside the Anoxomat. With the Anoxomat, anaerobiosis was achieved about 35 min faster than in the GasPak system. The density of growth recorded for 177 (78%) strains was heavier in the Anoxomat than in the GasPak jar. The zones of inhibition of the antibiotics tested were not different in the two systems. The Anoxomat system provided superior growth, in terms of density and colony size, and achieved anaerobiosis more rapidly. Evidently, the Anoxomat method is more reliable and appears to support the growth of strict anaerobes better. Copyright 2003 S. Karger AG, Basel

Decomposition of Alternative Chirality Amino Acids by Alkaliphilic Anaerobe from Owens Lake, California

Science.gov (United States)

Townsend, Alisa; Pikuta, Elena V.; Guisler, Melissa; Hoover, Richard B.

2009-01-01

The study of alkaliphilic microbial communities from anaerobic sediments of Owens and Mono Lakes in California led to the isolation of a bacterial strain capable of metabolizing amino acids with alternative chirality. According to the phylogenetic analysis, the anaerobic strain BK1 belongs to the genus Tindallia; however, despite the characteristics of other described species of this genus, the strain BK1 was able to grow on D-arginine and Dlysine. Cell morphology of this strain showed straight, motile, non-spore-forming rods with sizes 0.45 x 1.2-3 microns. Physiological characteristics of the strain showed that it is catalase negative, obligately anaerobic, mesophilic, and obligately alkaliphilic. This isolate is unable to grow at pH 7 and requires CO3 (2-) ions for growth. The strain has chemo-heterotrophic metabolism and is able to ferment various proteolysis products and some sugars. It plays the role of a primary anaerobe within the trophic chain of an anaerobic microbial community by the degradation of complex protein molecules to smaller and less energetic molecules. The new isolate requires NaCl for growth, and can grow within the range of 0.5-13 %, with the optimum at 1 % NaCl (w/v). The temperature range for the growth of the new isolate is 12-40 C with optimum at 35 C. The pH range for the growth of strain BK1 occurs between 7.8 and 11.0 with optimum at 9.5. This paper presents detailed physiological characteristics of the novel isolate from Owens Lake, a unique relic ecosystem of Astrobiological significance, and makes an accent on the ability of this strain to utilize L-amino acids.

Optimization of CDT-1 and XYL1 Expression for Balanced Co-Production of Ethanol and Xylitol from Cellobiose and Xylose by Engineered Saccharomyces cerevisiae

Science.gov (United States)

Zha, Jian; Li, Bing-Zhi; Shen, Ming-Hua; Hu, Meng-Long; Song, Hao; Yuan, Ying-Jin

2013-01-01

Production of ethanol and xylitol from lignocellulosic hydrolysates is an alternative to the traditional production of ethanol in utilizing biomass. However, the conversion efficiency of xylose to xylitol is restricted by glucose repression, causing a low xylitol titer. To this end, we cloned genes CDT-1 (encoding a cellodextrin transporter) and gh1-1 (encoding an intracellular β-glucosidase) from Neurospora crassa and XYL1 (encoding a xylose reductase that converts xylose into xylitol) from Scheffersomyces stipitis into Saccharomyces cerevisiae, enabling simultaneous production of ethanol and xylitol from a mixture of cellobiose and xylose (main components of lignocellulosic hydrolysates). We further optimized the expression levels of CDT-1 and XYL1 by manipulating their promoters and copy-numbers, and constructed an engineered S. cerevisiae strain (carrying one copy of PGK1p-CDT1 and two copies of TDH3p-XYL1), which showed an 85.7% increase in xylitol production from the mixture of cellobiose and xylose than that from the mixture of glucose and xylose. Thus, we achieved a balanced co-fermentation of cellobiose (0.165 g/L/h) and xylose (0.162 g/L/h) at similar rates to co-produce ethanol (0.36 g/g) and xylitol (1.00 g/g). PMID:23844185

Optimization of CDT-1 and XYL1 expression for balanced co-production of ethanol and xylitol from cellobiose and xylose by engineered Saccharomyces cerevisiae.

Directory of Open Access Journals (Sweden)

Jian Zha

Full Text Available Production of ethanol and xylitol from lignocellulosic hydrolysates is an alternative to the traditional production of ethanol in utilizing biomass. However, the conversion efficiency of xylose to xylitol is restricted by glucose repression, causing a low xylitol titer. To this end, we cloned genes CDT-1 (encoding a cellodextrin transporter and gh1-1 (encoding an intracellular β-glucosidase from Neurospora crassa and XYL1 (encoding a xylose reductase that converts xylose into xylitol from Scheffersomyces stipitis into Saccharomyces cerevisiae, enabling simultaneous production of ethanol and xylitol from a mixture of cellobiose and xylose (main components of lignocellulosic hydrolysates. We further optimized the expression levels of CDT-1 and XYL1 by manipulating their promoters and copy-numbers, and constructed an engineered S. cerevisiae strain (carrying one copy of PGK1p-CDT1 and two copies of TDH3p-XYL1, which showed an 85.7% increase in xylitol production from the mixture of cellobiose and xylose than that from the mixture of glucose and xylose. Thus, we achieved a balanced co-fermentation of cellobiose (0.165 g/L/h and xylose (0.162 g/L/h at similar rates to co-produce ethanol (0.36 g/g and xylitol (1.00 g/g.

Performance testing of Zymomonas mobilis metabolically engineered for cofermentation of glucose, xylose, and arabinose.

Science.gov (United States)

Lawford, Hugh G; Rousseau, Joyce D

2002-01-01

IOGEN Corporation of Ottawa, Canada, has recently built a 40t/d biomass-to-ethanol demonstration plant adjacent to its enzyme production facility. It has partnered with the University of Toronto to test the C6/C5 cofermenta-tion performance characteristics of the National Renewable Energy Labora-tory's metabolically engineered Zymomonas mobilis using various biomass hydrolysates. IOGEN's feedstocks are primarily agricultural wastes such as corn stover and wheat straw. Integrated recombinant Z. mobilis strain AX101 grows on D-xylose and/or L-arabinose as the sole carbon/energy sources and ferments these pentose sugars to ethanol in high yield. Strain AX101 lacks the tetracycline resistance gene that was a common feature of other recombinant Zm constructs. Genomic integration provides reliable cofermentation performance in the absence of antibiotics, another characteristic making strain AX101 attractive for industrial cellulosic ethanol production. In this work, IOGEN's biomass hydrolysate was simulated by a pure sugar medium containing 6% (w/v) glucose, 3% xylose, and 0.35% arabinose. At a level of 3 g/L (dry solids), corn steep liquor with inorganic nitrogen (0.8 g/L of ammonium chloride or 1.2 g/L of diammonium phosphate) was a cost-effective nutritional supplement. In the absence of acetic acid, the maximum volumetric ethanol productivity of a continuous fermentation at pH 5.0 was 3.54 g/L x h. During prolonged continuous fermentation, the efficiency of sugar-to-ethanol conversion (based on total sugar load) was maintained at >85%. At a level of 0.25% (w/v) acetic acid, the productivity decreased to 1.17 g/L x h at pH 5.5. Unlike integrated, xylose-utilizing rec Zm strain C25, strain AX101 produces less lactic acid as byproduct, owing to the fact that the Escherichia coli arabinose genes are inserted into a region of the host chromosome tentatively assigned to the gene for D-lactic acid dehydrogenase. In pH-controlled batch fermentations with sugar mixtures, the

Efficient Hydrolysis of Rice Straw into Xylose and Glucose by a Two-step Process

Directory of Open Access Journals (Sweden)

YAN Lu-lu

2016-07-01

Full Text Available The hydrolysis of rice straw into xylose and glucose in dilute sulfuric acid aqueous solution was studied with a two-step process in batch autoclave reactor. The results showed that compared with the traditional one-step acid hydrolysis, both xylose and glucose could be produced in high yields from rice straw by using the two-step acid hydrolysis process. The effects of reaction temperature, reaction time, the amount of rice straw and acid concentration on the hydrolysis of rice straw were systematically studied, and showed that except initial rice straw loading amount, the other parameters had remarkable influence on the products distribution and yields. In the first-step of the hydrolysis process, a high xylose yield of 162.6 g·kg-1 was obtained at 140℃ after 120 min reaction time. When the solid residues from the first step were subjected to a second-step hydrolysis, a glucose yield as high as 216.5 g·kg-1 could be achieved at 180℃ after 120 min. This work provides a promising strategy for the efficient and value-added utilization of agricultural wastes such as rice straw.

Xylose fermentation efficiency and inhibitor tolerance of the recombinant industrial Saccharomyces cerevisiae strain NAPX37.

Science.gov (United States)

Li, Yun-Cheng; Mitsumasu, Kanako; Gou, Zi-Xi; Gou, Min; Tang, Yue-Qin; Li, Guo-Ying; Wu, Xiao-Lei; Akamatsu, Takashi; Taguchi, Hisataka; Kida, Kenji

2016-02-01

Industrial yeast strains with good xylose fermentation ability and inhibitor tolerance are important for economical lignocellulosic bioethanol production. The flocculating industrial Saccharomyces cerevisiae strain NAPX37, harboring the xylose reductase-xylitol dehydrogenase (XR-XDH)-based xylose metabolic pathway, displayed efficient xylose fermentation during batch and continuous fermentation. During batch fermentation, the xylose consumption rates at the first 36 h were similar (1.37 g/L/h) when the initial xylose concentrations were 50 and 75 g/L, indicating that xylose fermentation was not inhibited even when the xylose concentration was as high as 75 g/L. The presence of glucose, at concentrations of up to 25 g/L, did not affect xylose consumption rate at the first 36 h. Strain NAPX37 showed stable xylose fermentation capacity during continuous ethanol fermentation using xylose as the sole sugar, for almost 1 year. Fermentation remained stable at a dilution rate of 0.05/h, even though the xylose concentration in the feed was as high as 100 g/L. Aeration rate, xylose concentration, and MgSO4 concentration were found to affect xylose consumption and ethanol yield. When the xylose concentration in the feed was 75 g/L, a high xylose consumption rate of 6.62 g/L/h and an ethanol yield of 0.394 were achieved under an aeration rate of 0.1 vvm, dilution rate of 0.1/h, and 5 mM MgSO4. In addition, strain NAPX37 exhibited good tolerance to inhibitors such as weak acids, furans, and phenolics during xylose fermentation. These findings indicate that strain NAPX37 is a promising candidate for application in the industrial production of lignocellulosic bioethanol.

Identification of a novel acetate-utilizing bacterium belonging to Synergistes group 4 in anaerobic digester sludge

OpenAIRE

Ito, Tsukasa; Yoshiguchi, Kazumi; Ariesyady, Herto Dwi; Okabe, Satoshi

2011-01-01

Major acetate-utilizing bacterial and archaeal populations in methanogenic anaerobic digester sludge were identified and quantified by radioisotope- and stable-isotope-based functional analyses, microautoradiography-fluorescence in situ hybridization (MAR-FISH) and stable-isotope probing of 16S rRNA (RNA-SIP) that can directly link 16S rRNA phylogeny with in situ metabolic function. First, MAR-FISH with 14C-acetate indicated the significant utilization of acetate by only two major groups, uni...

Genome-scale consequences of cofactor balancing in engineered pentose utilization pathways in Saccharomyces cerevisiae.

Directory of Open Access Journals (Sweden)

Amit Ghosh

Full Text Available Biofuels derived from lignocellulosic biomass offer promising alternative renewable energy sources for transportation fuels. Significant effort has been made to engineer Saccharomyces cerevisiae to efficiently ferment pentose sugars such as D-xylose and L-arabinose into biofuels such as ethanol through heterologous expression of the fungal D-xylose and L-arabinose pathways. However, one of the major bottlenecks in these fungal pathways is that the cofactors are not balanced, which contributes to inefficient utilization of pentose sugars. We utilized a genome-scale model of S. cerevisiae to predict the maximal achievable growth rate for cofactor balanced and imbalanced D-xylose and L-arabinose utilization pathways. Dynamic flux balance analysis (DFBA was used to simulate batch fermentation of glucose, D-xylose, and L-arabinose. The dynamic models and experimental results are in good agreement for the wild type and for the engineered D-xylose utilization pathway. Cofactor balancing the engineered D-xylose and L-arabinose utilization pathways simulated an increase in ethanol batch production of 24.7% while simultaneously reducing the predicted substrate utilization time by 70%. Furthermore, the effects of cofactor balancing the engineered pentose utilization pathways were evaluated throughout the genome-scale metabolic network. This work not only provides new insights to the global network effects of cofactor balancing but also provides useful guidelines for engineering a recombinant yeast strain with cofactor balanced engineered pathways that efficiently co-utilizes pentose and hexose sugars for biofuels production. Experimental switching of cofactor usage in enzymes has been demonstrated, but is a time-consuming effort. Therefore, systems biology models that can predict the likely outcome of such strain engineering efforts are highly useful for motivating which efforts are likely to be worth the significant time investment.

Expanding xylose metabolism in yeast for plant cell wall conversion to biofuels

Science.gov (United States)

Li, Xin; Yu, Vivian Yaci; Lin, Yuping; Chomvong, Kulika; Estrela, Raíssa; Park, Annsea; Liang, Julie M; Znameroski, Elizabeth A; Feehan, Joanna; Kim, Soo Rin; Jin, Yong-Su; Glass, N Louise; Cate, Jamie HD

2015-01-01

Sustainable biofuel production from renewable biomass will require the efficient and complete use of all abundant sugars in the plant cell wall. Using the cellulolytic fungus Neurospora crassa as a model, we identified a xylodextrin transport and consumption pathway required for its growth on hemicellulose. Reconstitution of this xylodextrin utilization pathway in Saccharomyces cerevisiae revealed that fungal xylose reductases act as xylodextrin reductases, producing xylosyl-xylitol oligomers as metabolic intermediates. These xylosyl-xylitol intermediates are generated by diverse fungi and bacteria, indicating that xylodextrin reduction is widespread in nature. Xylodextrins and xylosyl-xylitol oligomers are then hydrolyzed by two hydrolases to generate intracellular xylose and xylitol. Xylodextrin consumption using a xylodextrin transporter, xylodextrin reductases and tandem intracellular hydrolases in cofermentations with sucrose and glucose greatly expands the capacity of yeast to use plant cell wall-derived sugars and has the potential to increase the efficiency of both first-generation and next-generation biofuel production. DOI: http://dx.doi.org/10.7554/eLife.05896.001 PMID:25647728

Development of a strain of saccharomyces cereviase to utilize hemicellulosic biomass

International Nuclear Information System (INIS)

Batt, C.A.

1991-01-01

The current status of yeast conversion to utilize pentose sugar is discussed in this paper. The development of processes for the production of ethanol from agricultural wastes provides both a beneficial utilization of the resources presently available and an alternate source of liquid transportation fuel. The efficient conversion of agricultural bio mass is in part dependent on utilization of all the potential sugars, including the pentoses in the hemicellulosic fraction. A number of approaches have been investigated, including the engineering of strain of S. cerevisiae which express a xylose isomerase activity. Despite the apparent lack of success with respect to expressing an active xylose isomerase, a great deal of knowledge has been gained on the metabolism of pentoses by yeast and the genetics, structure/function of the enzyme xylose isomerase. Hopefully this cumulative knowledge base will lead to the design of a xylose isomerase with the appropriate structure to allow it retain activity in S. cerevisiae. This coupled with the elegant efforts in a number of laboratories to develop cellulose utilizing strains of S. cerevisiae might yield a single yeast capable of fermenting all of the major carbon substrates in agricultural to fuel grade ethanol. (Orig./A.B.)

Genomic sequence of the xylose fermenting, insect-inhabitingyeast, Pichia stipitis

Energy Technology Data Exchange (ETDEWEB)

Jeffries, Thomas W.; Grigoriev, Igor; Grimwood, Jane; Laplaza,Jose M.; Aerts, Andrea; Salamov, Asaf; Schmutz, Jeremy; Lindquist, Erika; Dehal, Paramvir; Shapiro, Harris; Jin, Yong-Su; Passoth, Volkmar; Richardson, Paul M.

2007-06-25

Xylose is a major constituent of angiosperm lignocellulose,so its fermentation is important for bioconversion to fuels andchemicals. Pichia stipitis is the best-studied native xylose fermentingyeast. Genes from P. stipitis have been used to engineer xylosemetabolism in Saccharomycescerevisiae, and the regulation of the P.stipitis genome offers insights into the mechanisms of xylose metabolismin yeasts. We have sequenced, assembled and finished the genome ofP.stipitis. As such, it is one of only a handful of completely finishedeukaryotic organisms undergoing analysis and manual curation. Thesequence has revealed aspects of genome organization, numerous genes forbiocoversion, preliminary insights into regulation of central metabolicpathways, numerous examples of co-localized genes with related functions,and evidence of how P. stipitis manages to achieve redox balance whilegrowing on xylose under microaerobic conditions.

Reaction mechanisms and kinetics of processing glucose, xylose and glucose-xylose mixtures under hot compressed water conditions for predicting bio-crude composition

DEFF Research Database (Denmark)

Grigoras, Ionela; Toor, Saqib Sohail; Rosendahl, Lasse Aistrup

Mechanisms for bio-crude formation during the conversion of glucose, xylose and glucose-xylose mixtures as biomass model compounds under hot compressed water conditions are investigated. Studies in literature have shown that the diverse products formed at the early stages of glucose or xylose...... conversion are 5-HMF, erythrose, glyceraldehyde, dihydroxyacetone, pyruvaldehyde, and saccharinic acids resulted through reactions such as dehydration, retro-aldol condensation and isomerization. However, these compounds are mostly water soluble compounds and lack the final steps towards formation of water...... insoluble components at longer reaction times. The effects of pressure, pH, catalyst and reaction time on the main products are examined thoroughly. The possible routes for the formation of oil compounds are developed....

Multi-stage Continuous Culture Fermentation of Glucose-Xylose Mixtures to Fuel Ethanol using Genetically Engineered Saccharomyces cerevisiae 424A

Science.gov (United States)

Multi-stage continuous (chemostat) culture fermentation (MCCF) with variable fermentor volumes was carried out to study utilizing glucose and xylose for ethanol production by means of mixed sugar fermentation (MSF). Variable fermentor volumes were used to enable enhanced sugar u...

Utilization of steam- and explosion-decompressed aspen wood by some anaerobes

Energy Technology Data Exchange (ETDEWEB)

Khan, A W; Asther, M; Giuliano, C

1984-01-01

Tests made to study the suitability of using steam- and explosion-decompressed aspen wood as a substrate in anaerobic fermentations indicated that after washing with dilute NaOH it becomes less than 80% accessible to both mesophilic and thermophilic cellulolytic anaerobes and cellulases, compared with delignified, ball-milled pulp. After washing, this material was also suitable for the single-step conversion of cellulose to EtOH using cocultures consisting of cellulolytic and EtOH-producing saccharolytic anaerobes; and without and after washing by the use of cellulolytic enzymes and ethanologenic anaerobes.

Gene expression cross-profiling in genetically modified industrial Saccharomyces cerevisiae strains during high-temperature ethanol production from xylose.

Science.gov (United States)

Ismail, Ku Syahidah Ku; Sakamoto, Takatoshi; Hatanaka, Haruyo; Hasunuma, Tomohisa; Kondo, Akihiko

2013-01-10

Production of ethanol from xylose at high temperature would be an economical approach since it reduces risk of contamination and allows both the saccharification and fermentation steps in SSF to be running at elevated temperature. Eight recombinant xylose-utilizing Saccharomyces cerevisiae strains developed from industrial strains were constructed and subjected to high-temperature fermentation at 38 °C. The best performing strain was sun049T, which produced up to 15.2 g/L ethanol (63% of the theoretical production), followed by sun048T and sun588T, both with 14.1 g/L ethanol produced. Via transcriptomic analysis, expression profiling of the top three best ethanol producing strains compared to a negative control strain, sun473T, led to the discovery of genes in common that were regulated in the same direction. Identification of the 20 most highly up-regulated and the 20 most highly down-regulated genes indicated that the cells regulate their central metabolism and maintain the integrity of the cell walls in response to high temperature. We also speculate that cross-protection in the cells occurs, allowing them to maintain ethanol production at higher concentration under heat stress than the negative controls. This report provides further transcriptomics information in the interest of producing a robust microorganism for high-temperature ethanol production utilizing xylose. Copyright © 2012 Elsevier B.V. All rights reserved.

Report on a survey in fiscal 1999. Survey on industrial utilization of microorganism reaction mechanisms under anaerobic condition; 1999 nendo kenki jokenka ni okeru biseibutsu hanno kiko no kogyoteki riyo ni kansuru chosa hokokusho

Energy Technology Data Exchange (ETDEWEB)

NONE

2000-03-01

Industrial utilization of reaction mechanisms of microorganisms under anaerobic condition permits structuring energy saving type production processes. The present survey has investigated features of new microorganisms under anaerobic condition and the status of researches thereon inside and outside the country, and discussed their future applications. Chapter 1 compares anaerobic microorganisms and functions of microorganism under anaerobic condition with those aerobic to describe their general features, and describes the purpose of this survey and the summary of the investigations. Chapter 2 surveys the current status of technologies to utilize microorganisms under anaerobic condition. Chapter 3 outlines metabolic characteristics of the anaerobic microorganisms, and extracts functions effective for material production by different anaerobic microorganisms to describe their applicability. Chapter 4 evaluates the system classification for the anaerobic microorganisms utilizing the basic arrangement of 16S rRNA genes, and extracts technical problems therein. Chapter 5 proposes structuring a total methane fermentation system including a raw material collecting process, and enhancing alcohol productivity of Zymomonas bacteria. (NEDO)

Ethanol production from xylose by recombinant Saccharomyces cerevisiae expressing protein-engineered NADH-preferring xylose reductase from Pichia stipitis.

Science.gov (United States)

Watanabe, Seiya; Abu Saleh, Ahmed; Pack, Seung Pil; Annaluru, Narayana; Kodaki, Tsutomu; Makino, Keisuke

2007-09-01

A recombinant Saccharomyces cerevisiae strain transformed with xylose reductase (XR) and xylitol dehydrogenase (XDH) genes from Pichia stipitis (PsXR and PsXDH, respectively) has the ability to convert xylose to ethanol together with the unfavourable excretion of xylitol, which may be due to intercellular redox imbalance caused by the different coenzyme specificity between NADPH-preferring XR and NAD(+)-dependent XDH. In this study, we focused on the effect(s) of mutated NADH-preferring PsXR in fermentation. The R276H and K270R/N272D mutants were improved 52- and 146-fold, respectively, in the ratio of NADH/NADPH in catalytic efficiency [(k(cat)/K(m) with NADH)/(k(cat)/K(m) with NADPH)] compared with the wild-type (WT), which was due to decrease of k(cat) with NADPH in the R276H mutant and increase of K(m) with NADPH in the K270R/N272D mutant. Furthermore, R276H mutation led to significant thermostabilization in PsXR. The most positive effect on xylose fermentation to ethanol was found by using the Y-R276H strain, expressing PsXR R276H mutant and PsXDH WT: 20 % increase of ethanol production and 52 % decrease of xylitol excretion, compared with the Y-WT strain expressing PsXR WT and PsXDH WT. Measurement of intracellular coenzyme concentrations suggested that maintenance of the of NADPH/NADP(+) and NADH/NAD(+) ratios is important for efficient ethanol fermentation from xylose by recombinant S. cerevisiae.

Densities, molar volumes, and isobaric expansivities of (d-xylose+hydrochloric acid+water) systems

International Nuclear Information System (INIS)

Zhang Qiufen; Yan Zhenning; Wang Jianji; Zhang Hucheng

2006-01-01

Densities of (d-xylose+HCl+water) have been measured at temperature in the range (278.15 to 318.15) K as a function of concentration of both d-xylose and hydrochloric acid. The densities have been used to estimate the molar volumes and isobaric expansivity of the ternary solutions. The molar volumes of the ternary solutions vary linearly with mole fraction of d-xylose. The standard partial molar volumes V 2,φ - bar for d-xylose in aqueous solutions of molality (0.2, 0.4, 0.7, 1.1, 1.6, and 2.1) mol.kg -1 HCl have been determined. In the investigated temperature range, the relation: V 2,φ - bar =c 1 +c 2 {(T/K)-273.15} 1/2 , can be used to describe the temperature dependence of the standard partial molar volumes. These results have, in conjunction with the results obtained in water, been used to deduce the standard volumes of transfer, Δ t V - bar , of d-xylose from water to aqueous HCl solutions. An increase in the transfer volume of d-xylose with increasing HCl concentrations has been explained by the stronger interactions of H + with the hydrophilic groups of d-xylose

Engineering industrial Saccharomyces cerevisiae strains for xylose fermentation and comparison for switchgrass conversion

Science.gov (United States)

Saccharomyces physiology and fermentation related properties vary broadly among industrial strains. In this study, six industrial strains of varied genetic background were engineered to ferment xylose. Aerobic growth rates on xylose were 0.040 h**-1 to 0.167 h**-1. Fermentation of xylose, glucose/xy...

Xylose-fermenting Pichia stipitis by genome shuffling for improved ethanol production.

Science.gov (United States)

Shi, Jun; Zhang, Min; Zhang, Libin; Wang, Pin; Jiang, Li; Deng, Huiping

2014-03-01

Xylose fermentation is necessary for the bioconversion of lignocellulose to ethanol as fuel, but wild-type Saccharomyces cerevisiae strains cannot fully metabolize xylose. Several efforts have been made to obtain microbial strains with enhanced xylose fermentation. However, xylose fermentation remains a serious challenge because of the complexity of lignocellulosic biomass hydrolysates. Genome shuffling has been widely used for the rapid improvement of industrially important microbial strains. After two rounds of genome shuffling, a genetically stable, high-ethanol-producing strain was obtained. Designated as TJ2-3, this strain could ferment xylose and produce 1.5 times more ethanol than wild-type Pichia stipitis after fermentation for 96 h. The acridine orange and propidium iodide uptake assays showed that the maintenance of yeast cell membrane integrity is important for ethanol fermentation. This study highlights the importance of genome shuffling in P. stipitis as an effective method for enhancing the productivity of industrial strains. © 2013 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

Utilization of biogas produced by anaerobic digestion of agro-industrial waste: Energy, economic and environmental effects.

Science.gov (United States)

Hublin, Andrea; Schneider, Daniel Rolph; Džodan, Janko

2014-07-01

Anaerobic digestion of agro-industrial waste is of significant interest in order to facilitate a sustainable development of energy supply. Using of material and energy potentials of agro-industrial waste, in the framework of technical, economic, and ecological possibilities, contributes in increasing the share of energy generated from renewable energy sources. The paper deals with the benefits arising from the utilization of biogas produced by co-digestion of whey and cow manure. The advantages of this process are the profitability of the plant and the convenience in realizing an anaerobic digestion plant to produce biogas that is enabled by the benefits from the sale of electric energy at favorable prices. Economic aspects are related to the capital cost (€ 2,250,000) of anaerobic digestion treatment in a biogas plant with a 300 kW power and 510 kW heating unit in a medium size farm (450 livestock units). Considering the optimum biogas yield of 20.7 dm(3) kg(-1) of wet substrate and methane content in the biogas obtained of 79%, the anaerobic process results in a daily methane production of 2,500 kg, with the maximum power generation of 2,160,000 kWh y(-1) and heat generation of 2,400,000 kWh y(-1) The net present value (NPV), internal rate of return (IRR) and payback period for implementation of profitable anaerobic digestion process is evaluated. Ecological aspects related to carbon dioxide (CO2) and methane (CH4) emission reduction are assessed. © The Author(s) 2014.

Improving the cyanide toxicity tolerance of anaerobic reactor: Microbial interactions and toxin reduction

International Nuclear Information System (INIS)

Gupta, Pragya; Ahammad, S.Z.; Sreekrishnan, T.R.

2016-01-01

Highlights: • Anaerobic batch study of 110 days. • Acclimatization for cyanide biodegradation. • Understanding inhibitory effects of cyanide on methane generation and VFA production. • Identification of microorganisms tolerant to cyanide. • Community analysis using DGGE and qPCR analyses. - Abstract: Anaerobic biological treatment of high organics containing wastewater is amongst the preferred treatment options but poor tolerance to toxins makes its use prohibitive. In this study, efforts have been made to understand the key parameters for developing anaerobic reactor, resilient to cyanide toxicity. A laboratory scale anaerobic batch reactor was set up to treat cyanide containing wastewater. The reactor was inoculated with anaerobic sludge obtained from a wastewater treatment plant and fresh cow dung in the ratio of 3:1. The focus was on acclimatization and development of cyanide-degrading biomass and to understand the toxic effects of cyanide on the dynamic equilibrium between various microbial groups. The sludge exposed to cyanide was found to have higher bacterial diversity than the control. It was observed that certain hydrogenotrophic methanogens and bacterial groups were able to grow and produce methane in the presence of cyanide. Also, it was found that hydrogen utilizing methanogens were more cyanide tolerant than acetate utilizing methanogens. So, effluents from various industries like electroplating, coke oven plant, petroleum refining, explosive manufacturing, and pesticides industries which are having high concentrations of cyanide can be treated by favoring the growth of the tolerant microbes in the reactors. It will provide much better treatment efficiency by overcoming the inhibitory effects of cyanide to certain extent.

Improving the cyanide toxicity tolerance of anaerobic reactor: Microbial interactions and toxin reduction

Energy Technology Data Exchange (ETDEWEB)

Gupta, Pragya; Ahammad, S.Z.; Sreekrishnan, T.R., E-mail: sree@iitd.ac.in

2016-09-05

Highlights: • Anaerobic batch study of 110 days. • Acclimatization for cyanide biodegradation. • Understanding inhibitory effects of cyanide on methane generation and VFA production. • Identification of microorganisms tolerant to cyanide. • Community analysis using DGGE and qPCR analyses. - Abstract: Anaerobic biological treatment of high organics containing wastewater is amongst the preferred treatment options but poor tolerance to toxins makes its use prohibitive. In this study, efforts have been made to understand the key parameters for developing anaerobic reactor, resilient to cyanide toxicity. A laboratory scale anaerobic batch reactor was set up to treat cyanide containing wastewater. The reactor was inoculated with anaerobic sludge obtained from a wastewater treatment plant and fresh cow dung in the ratio of 3:1. The focus was on acclimatization and development of cyanide-degrading biomass and to understand the toxic effects of cyanide on the dynamic equilibrium between various microbial groups. The sludge exposed to cyanide was found to have higher bacterial diversity than the control. It was observed that certain hydrogenotrophic methanogens and bacterial groups were able to grow and produce methane in the presence of cyanide. Also, it was found that hydrogen utilizing methanogens were more cyanide tolerant than acetate utilizing methanogens. So, effluents from various industries like electroplating, coke oven plant, petroleum refining, explosive manufacturing, and pesticides industries which are having high concentrations of cyanide can be treated by favoring the growth of the tolerant microbes in the reactors. It will provide much better treatment efficiency by overcoming the inhibitory effects of cyanide to certain extent.

Continuous Ethanol Fermentation of Pretreated Lignocellulosic Biomasses, Waste Biomasses, Molasses and Syrup Using the Anaerobic, Thermophilic Bacterium Thermoanaerobacter italicus Pentocrobe 411

Science.gov (United States)

Andersen, Rasmus Lund; Jensen, Karen Møller; Mikkelsen, Marie Just

2015-01-01

Lignocellosic ethanol production is now at a stage where commercial or semi-commercial plants are coming online and, provided cost effective production can be achieved, lignocellulosic ethanol will become an important part of the world bio economy. However, challenges are still to be overcome throughout the process and particularly for the fermentation of the complex sugar mixtures resulting from the hydrolysis of hemicellulose. Here we describe the continuous fermentation of glucose, xylose and arabinose from non-detoxified pretreated wheat straw, birch, corn cob, sugar cane bagasse, cardboard, mixed bio waste, oil palm empty fruit bunch and frond, sugar cane syrup and sugar cane molasses using the anaerobic, thermophilic bacterium Thermoanaerobacter Pentocrobe 411. All fermentations resulted in close to maximum theoretical ethanol yields of 0.47–0.49 g/g (based on glucose, xylose, and arabinose), volumetric ethanol productivities of 1.2–2.7 g/L/h and a total sugar conversion of 90–99% including glucose, xylose and arabinose. The results solidify the potential of Thermoanaerobacter strains as candidates for lignocellulose bioconversion. PMID:26295944

Design of Xylose-Based Semisynthetic Polyurethane Tissue Adhesives with Enhanced Bioactivity Properties.

Science.gov (United States)

Balcioglu, Sevgi; Parlakpinar, Hakan; Vardi, Nigar; Denkbas, Emir Baki; Karaaslan, Merve Goksin; Gulgen, Selam; Taslidere, Elif; Koytepe, Suleyman; Ates, Burhan

2016-02-01

Developing biocompatible tissue adhesives with high adhesion properties is a highly desired goal of the tissue engineering due to adverse effects of the sutures. Therefore, our work involves synthesis, characterization, adhesion properties, protein adsorption, in vitro biodegradation, in vitro and in vivo biocompatibility properties of xylose-based semisynthetic polyurethane (NPU-PEG-X) bioadhesives. Xylose-based semisynthetic polyurethanes were developed by the reaction among 4,4'-methylenebis(cyclohexyl isocyanate) (MCI), xylose and polyethylene glycol 200 (PEG). Synthesized polyurethanes (PUs) showed good thermal stability and high adhesion strength. The highest values in adhesion strength were measured as 415.0 ± 48.8 and 94.0 ± 2.8 kPa for aluminum substrate and muscle tissue in 15% xylose containing PUs (NPU-PEG-X-15%), respectively. The biodegradation of NPU-PEG-X-15% was also determined as 19.96 ± 1.04% after 8 weeks of incubation. Relative cell viability of xylose containing PU was above 86%. Moreover, 10% xylose containing NPU-PEG-X (NPU-PEG-X-10%) sample has favorable tissue response, and inflammatory reaction between 1 and 6 weeks implantation period. With high adhesiveness and biocompatibility properties, NPU-PEG-X can be used in the medical field as supporting materials for preventing the fluid leakage after abdominal surgery or wound closure.

The production of anaerobic bacteria and biogas from dairy cattle waste in various growth mediums

Science.gov (United States)

Hidayati, Y. A.; Kurnani, T. B. A.; Marlina, E. T.; Rahmah, K. N.; Harlia, E.; Joni, I. M.

2018-02-01

The growth of anaerobic bacteria except the ruminal fluid quailty is strongly influenced by the media formulations. Previous researchers have set a standard media formulation for anaerobic bacteria from rumen, however the use of standard media formulations require chemicals with high cost. Moreover, other constraint of using standard media formulations is requires large quantities of media for anaerobic bacteria to grow. Therefore, it is necessary to find media with a new culture media formulation. Media used in this research were minimalist media consist of Nutrient Agar (NA), Lactose broth and rumen fluid; enriched media Rumen Fluid-Glucose-Agar (RGCA); and enriched media 98-5. The dairy cattle waste is utilized as source of anaerobic bacteria. The obtained data was analyzed by descriptive approach. The results showed that minimalist media produced anaerobic bacteria 2148 × 104 cfu/ml and biogas production: 1.06% CH4, 9.893% CO2; enriched media Rumen Fluid-Glucose-Agar (RGCA) produced anaerobic bacteria 1848 × 104 cfu/ml and biogas production 4.644% CH4, 9.5356% CO2; enriched media 98-5 produced anaerobic bacteria growth 15400 × 104 cfu/ml and biogas production 0.83% of CH4, 42.2% of CO2. It is conclude that the minimalist media was showed the best performance for the dairy cattle waste as source of anaerobic bacteria.

Effects of anaerobic growth conditions on biomass accumulation, root morphology, and efficiencies of nutrient uptake and utilization in seedlings of some southern coastal plain pine species

International Nuclear Information System (INIS)

Topa, M.A.

1984-01-01

Seedlings of pond (Pinus serotina (Michx.)), sand (P. clausa (Engelm.) Sarg.), and loblolly pines (P. taeda L., drought-hardy and wet site seed sources) were grown in a non-circulating, continuously-flowing solution culture under anaerobic or aerobic conditions to determine the effects of anaerobics on overall growth, root morphology and efficiencies of nutrient uptake and utilization. Although shoot growth of the 11-week old loblolly and pond pines was not affected by anaerobic treatment, it did significantly reduce root biomass. Sand pine suffered the largest biomass reduction. Flooding tolerance was positively correlated with specific morphological changes which enhanced root internal aeration. Oxygen transport from shoot to the root in anaerobically-grown loblolly and pond pine seedlings was demonstrated via rhizosphere oxidation experiments. Tissue elemental analyses showed that anaerobic conditions interfered with nutrient absorption and utilization. Short-term 32 p uptake experiments with intact seedlings indicated that net absorption decreased because of the reduction in root biomass, since H 2 PO 4 - influx in the anaerobically-grown seedlings was more than twice that of their aerobic counterparts. Sand pine possessed the physiological but not morphological capacity to increase P uptake under anaerobic growth conditions. Pond and wet-site loblolly pine seedlings maintained root growth, perhaps through enhanced internal root aeration - an advantage in field conditions where the phosphorus supply may be limited or highly localized

A novel method to prepare L-Arabinose from xylose mother liquor by yeast-mediated biopurification

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Lin Shuangjun

2011-06-01

Full Text Available Abstract Background L-arabinose is an important intermediate for anti-virus drug synthesis and has also been used in food additives for diets-controlling in recent years. Commercial production of L-arabinose is a complex progress consisting of acid hydrolysis of gum arabic, followed by multiple procedures of purification, thus making high production cost. Therefore, there is a biotechnological and commercial interest in the development of new cost-effective and high-performance methods for obtaining high purity grade L-arabinose. Results An alternative, economical method for purifying L-arabinose from xylose mother liquor was developed in this study. After screening 306 yeast strains, a strain of Pichia anomala Y161 was selected as it could effectively metabolize other sugars but not L-arabinose. Fermentation in a medium containing xylose mother liquor permitted enrichment of L-arabinose by a significant depletion of other sugars. Biochemical analysis of this yeast strain confirmed that its poor capacity for utilizing L-arabinose was due to low activities of the enzymes required for the metabolism of this sugar. Response surface methodology was employed for optimization the fermentation conditions in shake flask cultures. The optimum conditions were: 75 h fermentation time, at 32.5°C, in a medium containing 21% (v/v xylose mother liquor. Under these conditions, the highest purity of L-arabinose reached was 86.1% of total sugar, facilitating recovery of white crystalline L-arabinose from the fermentation medium by simple methods. Conclusion Yeast-mediated biopurification provides a dynamic method to prepare high purity of L-arabinose from the feedstock solution xylose mother liqour, with cost-effective and high-performance properties.

Xylose Isomerization with Zeolites in a Two-Step Alcohol–Water Process

DEFF Research Database (Denmark)

Paniagua, Marta; Shunmugavel, Saravanamurugan; Melián Rodriguez, Mayra

2015-01-01

Isomerization of xylose to xylulose was efficiently catalyzed by large-pore zeolites in a two-step methanol–water process that enhanced the product yield significantly. The reaction pathway involves xylose isomerization to xylulose, which, in part, subsequently reacts with methanol to form methyl...

Characterization and detection of a widely distributed gene cluster that predicts anaerobic choline utilization by human gut bacteria.

Science.gov (United States)

Martínez-del Campo, Ana; Bodea, Smaranda; Hamer, Hilary A; Marks, Jonathan A; Haiser, Henry J; Turnbaugh, Peter J; Balskus, Emily P

2015-04-14

Elucidation of the molecular mechanisms underlying the human gut microbiota's effects on health and disease has been complicated by difficulties in linking metabolic functions associated with the gut community as a whole to individual microorganisms and activities. Anaerobic microbial choline metabolism, a disease-associated metabolic pathway, exemplifies this challenge, as the specific human gut microorganisms responsible for this transformation have not yet been clearly identified. In this study, we established the link between a bacterial gene cluster, the choline utilization (cut) cluster, and anaerobic choline metabolism in human gut isolates by combining transcriptional, biochemical, bioinformatic, and cultivation-based approaches. Quantitative reverse transcription-PCR analysis and in vitro biochemical characterization of two cut gene products linked the entire cluster to growth on choline and supported a model for this pathway. Analyses of sequenced bacterial genomes revealed that the cut cluster is present in many human gut bacteria, is predictive of choline utilization in sequenced isolates, and is widely but discontinuously distributed across multiple bacterial phyla. Given that bacterial phylogeny is a poor marker for choline utilization, we were prompted to develop a degenerate PCR-based method for detecting the key functional gene choline TMA-lyase (cutC) in genomic and metagenomic DNA. Using this tool, we found that new choline-metabolizing gut isolates universally possessed cutC. We also demonstrated that this gene is widespread in stool metagenomic data sets. Overall, this work represents a crucial step toward understanding anaerobic choline metabolism in the human gut microbiota and underscores the importance of examining this microbial community from a function-oriented perspective. Anaerobic choline utilization is a bacterial metabolic activity that occurs in the human gut and is linked to multiple diseases. While bacterial genes responsible for

Tandem mass spectrometric characterization of the conversion of xylose to furfural

International Nuclear Information System (INIS)

Vinueza, Nelson R.; Kim, Eurick S.; Gallardo, Vanessa A.; Mosier, Nathan S.; Abu-Omar, Mahdi M.; Carpita, Nicholas C.; Kenttämaa, Hilkka I.

2015-01-01

Thermal decomposition of xylose into furfural under acidic conditions has been studied using tandem mass spectrometry. Two different Brønsted acids, maleic and sulfuric acids, were used to demonstrate that varying the Brønsted acid does not affect the mechanism of the reaction. Two selectively labeled xylose molecules, 1- 13 C and 5- 13 C-xyloses, were examined to determine which carbon atom is converted to the aldehyde carbon in furfural. This can be done by using tandem mass spectrometry since collision-activated dissociation (CAD) of protonated unlabeled furfural results in the loss of CO from the aldehyde moiety. The loss of a neutral molecule with MW of 29 Da ( 13 CO) was observed for protonated furfural derived from 1- 13 C-labeled xylose while the loss of a neutral molecule with MW of 28 Da (CO) was observed for protonated furfural derived from 5- 13 C labeled xylose. These results support the hypothesis that the mechanism of formation of furfural under mildly hot acidic conditions involves an intramolecular rearrangement of protonated xylose into the pyranose form rather than into an open-chain form. - Highlights: • Mechanism of catalytic conversion of Xyl to furfural under acidic conditions was studied by MS/MS and partially labeled Xyl. • The type of acid does not have a strong influence on the mechanism of catalytic conversion of Xyl to furfural. • The mechanism of formation of furfural under mildly hot acidic conditions involves an intramolecular rearrangement of Xyl

Pretreatment of wheat straw and conversion of xylose and xylan to ethanol by thermophilic anaerobic bacteria

DEFF Research Database (Denmark)

Ahring, Birgitte Kiær; Jensen, K.; Nielsen, P.

1996-01-01

solubilization was investigated. The two process parameters had little effect on the solubilization of hemicellulose. However alkaline conditions affected the furfural formation whereas oxygen had no effect. After pretreatment, the filtrate was used as a fermentation medium for thermophilic anaerobic bacterin...

Analysis of anaerobic blood cultures in burned patients.

Science.gov (United States)

Regules, Jason A; Carlson, Misty D; Wolf, Steven E; Murray, Clinton K

2007-08-01

The utility of anaerobic blood culturing is often debated in the general population, but there is limited data on the modern incidence, microbiology, and utility of obtaining routine anaerobic blood cultures for burned patients. We performed a retrospective review of the burned patients electronic medical records database for all blood cultures drawn between January 1997 and September 2005. We assessed blood cultures for positivity, organisms identified, and growth in aerobic or anaerobic media. 85,103 blood culture sets were drawn, with 4059 sets from burned patients. Three hundred and forty-five single species events (619 total blood culture isolates) were noted in 240 burned patients. For burned patients, four isolates were obligate anaerobic bacteria (all Propionibacterium acnes). Anaerobic versus aerobic culture growth was recorded in 310 of 619 (50.1%) burned patient blood culture sets. 46 (13.5%) of the identified organisms, most of which were not obligate anaerobic bacteria, were identified from solely anaerobic media. The results of our study suggest that the detection of significant anaerobic bacteremia in burned patients is very rare and that anaerobic bottles are not needed in this population for that indication. However anaerobic blood cultures systems are also able to detect facultative and obligate aerobic bacteria; therefore, the deletion of the anaerobic culture medium may have deleterious clinical impact.

Optimized Production of Xylitol from Xylose Using a Hyper-Acidophilic Candida tropicalis.

Science.gov (United States)

Tamburini, Elena; Costa, Stefania; Marchetti, Maria Gabriella; Pedrini, Paola

2015-08-19

The yeast Candida tropicalis DSM 7524 produces xylitol, a natural, low-calorie sweetener, by fermentation of xylose. In order to increase xylitol production rate during the submerged fermentation process, some parameters-substrate (xylose) concentration, pH, aeration rate, temperature and fermentation strategy-have been optimized. The maximum xylitol yield reached at 60-80 g/L initial xylose concentration, pH 5.5 at 37 °C was 83.66% (w/w) on consumed xylose in microaerophilic conditions (kLa = 2·h(-1)). Scaling up on 3 L fermenter, with a fed-batch strategy, the best xylitol yield was 86.84% (w/w), against a 90% of theoretical yield. The hyper-acidophilic behaviour of C. tropicalis makes this strain particularly promising for industrial application, due to the possibility to work in non-sterile conditions.

A Novel Technique that Enables Efficient Conduct of Simultaneous Isomerization and Fermentation (SIF) of Xylose

Science.gov (United States)

Rao, Kripa; Chelikani, Silpa; Relue, Patricia; Varanasi, Sasidhar

Of the sugars recovered from lignocellulose, D-glucose can be readily converted into ethanol by baker's or brewer's yeast (Saccharomyces cerevisiae). However, xylose that is obtained by the hydrolysis of the hemicellulosic portion is not fermentable by the same species of yeasts. Xylose fermentation by native yeasts can be achieved via isomerization of xylose to its ketose isomer, xylulose. Isomerization with exogenous xylose isomerase (XI) occurs optimally at a pH of 7-8, whereas subsequent fermentation of xylulose to ethanol occurs at a pH of 4-5. We present a novel scheme for efficient isomerization of xylose to xylulose at conditions suitable for the fermentation by using an immobilized enzyme system capable of sustaining two different pH microenvironments in a single vessel. The proof-of-concept of the two-enzyme pellet is presented, showing conversion of xylose to xylulose even when the immobilized enzyme pellets are suspended in a bulk solution whose pH is sub-optimal for XI activity. The co-immobilized enzyme pellets may prove extremely valuable in effectively conducting "simultaneous isomerization and fermentation" (SIF) of xylose. To help further shift the equilibrium in favor of xylulose formation, sodium tetraborate (borax) was added to the isomerization solution. Binding of tetrahydroxyborate ions to xylulose effectively reduces the concentration of xylulose and leads to increased xylose isomerization. The formation of tetrahydroxyborate ions and the enhancement in xylulose production resulting from the complexation was studied at two different bulk pH values. The addition of 0.05 M borax to the isomerization solution containing our co-immobilized enzyme pellets resulted in xylose to xylulose conversion as high as 86% under pH conditions that are suboptimal for XI activity. These initial findings, which can be optimized for industrial conditions, have significant potential for increasing the yield of ethanol from xylose in an SIF approach.

Separation of xylose oligomers using centrifugal partition chromatography with a butanol-methanol-water system.

Science.gov (United States)

Lau, Ching-Shuan; Clausen, Edgar C; Lay, Jackson O; Gidden, Jennifer; Carrier, Danielle Julie

2013-01-01

Xylose oligomers are the intermediate products of xylan depolymerization into xylose monomers. An understanding of xylan depolymerization kinetics is important to improve the conversion of xylan into monomeric xylose and to minimize the formation of inhibitory products, thereby reducing ethanol production costs. The study of xylan depolymerization requires copious amount of xylose oligomers, which are expensive if acquired commercially. Our approach consisted of producing in-house oligomer material. To this end, birchwood xylan was used as the starting material and hydrolyzed in hot water at 200 °C for 60 min with a 4 % solids loading. The mixture of xylose oligomers was subsequently fractionated by a centrifugal partition chromatography (CPC) with a solvent system of butanol:methanol:water in a 5:1:4 volumetric ratio. Operating in an ascending mode, the butanol-rich upper phase (the mobile phase) eluted xylose oligomers from the water-rich stationary phase at a 4.89 mL/min flow rate for a total fractionation time of 300 min. The elution of xylose oligomers occurred between 110 and 280 min. The yields and purities of xylobiose (DP 2), xylotriose (DP 3), xylotetraose (DP 4), and xylopentaose (DP 5) were 21, 10, 14, and 15 mg/g xylan and 95, 90, 89, and 68 %, respectively. The purities of xylose oligomers from this solvent system were higher than those reported previously using tetrahydrofuran:dimethyl sulfoxide:water in a 6:1:3 volumetric ratio. Moreover, the butanol-based solvent system improved overall procedures by facilitating the evaporation of the solvents from the CPC fractions, rendering the purification process more efficient.

Furfural and glucose can enhance conversion of xylose to xylitol by Candida magnoliae TISTR 5663.

Science.gov (United States)

Wannawilai, Siwaporn; Lee, Wen-Chien; Chisti, Yusuf; Sirisansaneeyakul, Sarote

2017-01-10

Xylitol production from xylose by the yeast Candida magnoliae TISTR 5663 was enhanced by supplementing the fermentation medium with furfural (300mg/L) and glucose (3g/L with an initial mass ratio of glucose to xylose of 1:10) together under oxygen limiting conditions. In the presence of furfural and glucose, the final concentration of xylitol was unaffected relative to control cultures but the xylitol yield on xylose increased by about 5%. Supplementation of the culture medium with glucose alone at an initial concentration of 3g/L, stimulated the volumetric and specific rates of xylose consumption and the rate of xylitol production from xylose. In a culture medium containing 30g/L xylose, 300mg/L furfural and 3g/L glucose, the volumetric production rate of xylitol was 1.04g/L h and the specific production rate was 0.169g/g h. In the absence of furfural and glucose, the volumetric production rate of xylitol was ∼35% lower and the specific production rate was nearly 30% lower. In view of these results, xylose-containing lignocellulosic hydrolysates contaminated with furfural can be effectively used for producing xylitol by fermentation so long as the glucose-to-xylose mass ratio in the hydrolysate does not exceed 1:10 and the furfural concentration is ≤300mg/L. Copyright © 2016 Elsevier B.V. All rights reserved.

Isolation and characterization of yeasts capable of efficient utilization of hemicellulosic hydrolyzate as the carbon source.

Science.gov (United States)

Cassa-Barbosa, L A; Procópio, R E L; Matos, I T S R; Filho, S A

2015-09-28

Few yeasts have shown the potential to efficiently utilize hemicellulosic hydrolyzate as the carbon source. In this study, microorganisms isolated from the Manaus region in Amazonas, Brazil, were characterized based on their utilization of the pentoses, xylose, and arabinose. The yeasts that showed a potential to assimilate these sugars were selected for the better utilization of lignocellulosic biomass. Two hundred and thirty seven colonies of unicellular microorganisms grown on hemicellulosic hydrolyzate, xylose, arabinose, and yeast nitrogen base selective medium were analyzed. Of these, 231 colonies were subjected to sugar assimilation tests. One hundred and twenty five of these were shown to utilize hydrolyzed hemicellulose, xylose, or arabinose as the carbon source for growth. The colonies that showed the best growth (N = 57) were selected, and their internal transcribed spacer-5.8S rDNA was sequenced. The sequenced strains formed four distinct groups in the phylogenetic tree, and showed a high percentage of similarity with Meyerozyma caribbica, Meyerozyma guilliermondii, Trichosporon mycotoxinivorans, Trichosporon loubieri, Pichia kudriavzevii, Candida lignohabitans, and Candida ethanolica. The discovery of these xylose-fermenting yeasts could attract widespread interest, as these can be used in the cost-effective production of liquid fuel from lignocellulosic materials.

[Isolation and characterization of Thermopirellula anaerolimosa gen. nov., sp. nov., an obligate anaerobic hydrogen-producing bacterium of the phylum Planctomycetes].

Science.gov (United States)

Liu, Dongying; Liu, Yi; Men, Xuehui; Guo, Qunqun; Guo, Rongbo; Qiu, Yanling

2012-08-04

To cultivate various yet-to-be cultured heterotrophs from anaerobic granule sludge, we used a selective culture medium with low concentrations of substrates supplemented a variety of antibiotics. An obligate anaerobic, thermophilic, hydrogen-producing bacterium, strain VM20-7(T), was isolated from an upflow anaerobic sludge blanket (UASB) reactor treating high-strength organic wastewater from isomerized sugar production processes. Cells of strain VM20-7(T) are non-motile, spherical, pear or teardrop shaped, occurring singly(o)r as aggregates (0.7 - 2.0 microm x 0.7 - 2.0 microm). Spore formation was not observed. Growth temperature ranges from 35 - 50 degrees C (optimum 45 degrees C), pH ranges from 6.0 - 8.3 (optimum 7.0 - 7.5) , NaCl tolerant concentration ranges from 0% - 0.5% (w/v, optimum 0% ). Nitrate, sulfate, thiosulfate, sulfite, elemental sulfur and Fe (III)-NTA were not used as terminal electron acceptors. Strain VM20-7(T) utilizes a wide range of carbohydrates, including glucose, maltose, ribose, xylose, sucrose, galactose, mannose, raffinose, pectin, yeast extract and xylan. Acetate and H2 are the main end products of glucose fermentation. The G + C content of the genomic DNA was 60.9 mol%. 16S rRNA gene sequence analysis revealed that it is related to the Pirellula-Rhodopirellula-Blastopirellula (PRB) clade within the order Planctomycetales (82.7 - 84.3% similarity with 16S rRNA genes of other known related species). The first obligate anaerobic bacterium within the phylum Planctomycetes was isolated with low concentration of carbohydrates and antibiotics. On the basis of the physiological and phylogenetic data, the name Thermopirellula anaerolimosa gen. nov. , sp. nov. is proposed for strain VM20-7(T) (= CGMCC 1.5169(T) = JCM 17478(T) = DSM 24165(T)).

Optimized Production of Xylitol from Xylose Using a Hyper-Acidophilic Candida tropicalis

Directory of Open Access Journals (Sweden)

Elena Tamburini

2015-08-01

Full Text Available The yeast Candida tropicalis DSM 7524 produces xylitol, a natural, low-calorie sweetener, by fermentation of xylose. In order to increase xylitol production rate during the submerged fermentation process, some parameters-substrate (xylose concentration, pH, aeration rate, temperature and fermentation strategy-have been optimized. The maximum xylitol yield reached at 60–80 g/L initial xylose concentration, pH 5.5 at 37 °C was 83.66% (w/w on consumed xylose in microaerophilic conditions (kLa = 2·h−1. Scaling up on 3 L fermenter, with a fed-batch strategy, the best xylitol yield was 86.84% (w/w, against a 90% of theoretical yield. The hyper-acidophilic behaviour of C. tropicalis makes this strain particularly promising for industrial application, due to the possibility to work in non-sterile conditions.

Xylose donor transport is critical for fungal virulence.

Directory of Open Access Journals (Sweden)

Lucy X Li

2018-01-01

Full Text Available Cryptococcus neoformans, an AIDS-defining opportunistic pathogen, is the leading cause of fungal meningitis worldwide and is responsible for hundreds of thousands of deaths annually. Cryptococcal glycans are required for fungal survival in the host and for pathogenesis. Most glycans are made in the secretory pathway, although the activated precursors for their synthesis, nucleotide sugars, are made primarily in the cytosol. Nucleotide sugar transporters are membrane proteins that solve this topological problem, by exchanging nucleotide sugars for the corresponding nucleoside phosphates. The major virulence factor of C. neoformans is an anti-phagocytic polysaccharide capsule that is displayed on the cell surface; capsule polysaccharides are also shed from the cell and impede the host immune response. Xylose, a neutral monosaccharide that is absent from model yeast, is a significant capsule component. Here we show that Uxt1 and Uxt2 are both transporters specific for the xylose donor, UDP-xylose, although they exhibit distinct subcellular localization, expression patterns, and kinetic parameters. Both proteins also transport the galactofuranose donor, UDP-galactofuranose. We further show that Uxt1 and Uxt2 are required for xylose incorporation into capsule and protein; they are also necessary for C. neoformans to cause disease in mice, although surprisingly not for fungal viability in the context of infection. These findings provide a starting point for deciphering the substrate specificity of an important class of transporters, elucidate a synthetic pathway that may be productively targeted for therapy, and contribute to our understanding of fundamental glycobiology.

D-xylose test of resorption as a method to determine radiation side effects in small intestine

International Nuclear Information System (INIS)

Koest, S.; Keinert, K.; Glaser, F.H.

1998-01-01

Background: The D-xylose test is the most important method to determine a disorder of carbohydrates resorption in proximal small intestine. The application is based on an impaired resorption due to pathological change of small intestine surface, leading to a decreased blood level or decreased excretion in urine. Patients and Method: D-xylose test was applied in 91 patients before, shortly after, 1/2 and 1 year after radiotherapy. All patients received an abdominal radiotherapy. We determined the blood level of D-xylose by a capillary blood sample 1 hour after oral D-xylose administration. Results: A significant decrease of the mean blood level of D-xylose to 1.88 mmol/l was determined after radiotherapy in comparison with 2.17 mmol/l before radiotherapy. Half a year after radiotherapy the mean blood level of D-xylose returned to normal. Regarding a threshold value of D-xylose blood level of 1.70 mmol/l 29 patients (32%) showed a pathologically decreased D-xylose resorption after radiotherapy. Twenty out of the 29 patients already showed a normal resorption half a year after the determination of the resorption disorder, 5 patients after 1 year and 4 patients after 1 1/2 years. There was no correlation between the detection of a disorder of D-xylose resorption and of a loss of body weight. The acute clinical side effects seemed to be more marked in connection with a disorder of D-xylose resorption, but this correlation is not significant. Eleven or 14 of the 29 patients, respectively, with pathologically decreased D-xylose resorption only had complaints of lower or upper gastrointestinal tract, respectively, and 10 patients did not have abdominal complaints at all. Conclusions: The D-xylose test is an important and simple method for determination of radiogen induced carbohydrate malabsorption in proximal small intestine. By means of its radiation side effects on small intestine can also be determined in patients who are otherwise free of complaints. (orig.) [de

Improved Ethanol Production from Xylose by Candida shehatae Induced by Dielectric Barrier Discharge Air Plasma

International Nuclear Information System (INIS)

Chen Huixia; Xiu Zhilong; Bai Fengwu

2014-01-01

Xylose fermentation is essential for ethanol production from lignocellulosic biomass. Exposure of the xylose-fermenting yeast Candida shehatae (C. shehatae) CICC1766 to atmospheric pressure dielectric barrier discharge (DBD) air plasma yields a clone (designated as C81015) with stability, which exhibits a higher ethanol fermentation rate from xylose, giving a maximal enhancement in ethanol production of 36.2% compared to the control (untreated). However, the biomass production of C81015 is lower than that of the control. Analysis of the NADH (nicotinamide adenine dinucleotide)- and NADPH (nicotinamide adenine dinucleotide phosphate)-linked xylose reductases and NAD + -linked xylitol dehydrogenase indicates that their activities are enhanced by 34.1%, 61.5% and 66.3%, respectively, suggesting that the activities of these three enzymes are responsible for improving ethanol fermentation in C81015 with xylose as a substrate. The results of this study show that DBD air plasma could serve as a novel and effective means of generating microbial strains that can better use xylose for ethanol fermentation

Improved Ethanol Production from Xylose by Candida shehatae Induced by Dielectric Barrier Discharge Air Plasma

Science.gov (United States)

Chen, Huixia; Xiu, Zhilong; Bai, Fengwu

2014-06-01

Xylose fermentation is essential for ethanol production from lignocellulosic biomass. Exposure of the xylose-fermenting yeast Candida shehatae (C. shehatae) CICC1766 to atmospheric pressure dielectric barrier discharge (DBD) air plasma yields a clone (designated as C81015) with stability, which exhibits a higher ethanol fermentation rate from xylose, giving a maximal enhancement in ethanol production of 36.2% compared to the control (untreated). However, the biomass production of C81015 is lower than that of the control. Analysis of the NADH (nicotinamide adenine dinucleotide)- and NADPH (nicotinamide adenine dinucleotide phosphate)-linked xylose reductases and NAD+-linked xylitol dehydrogenase indicates that their activities are enhanced by 34.1%, 61.5% and 66.3%, respectively, suggesting that the activities of these three enzymes are responsible for improving ethanol fermentation in C81015 with xylose as a substrate. The results of this study show that DBD air plasma could serve as a novel and effective means of generating microbial strains that can better use xylose for ethanol fermentation.

Direct production of D-arabinose from D-xylose by a coupling reaction using D-xylose isomerase, D-tagatose 3-epimerase and D-arabinose isomerase.

Science.gov (United States)

Sultana, Ishrat; Mizanur, Rahman Md; Takeshita, Kei; Takada, Goro; Izumori, Ken

2003-01-01

Klebsiella pneumoniae 40bXX, a mutant strain that constitutively produces D-arabinose isomerase (D-AI), was isolated through a series of repeated subcultures from the parent strain on a mineral salt medium supplemented with L-Xylose as the sole carbon source. D-AI could be efficiently immobilized on chitopearl beads. The optimum temperature for the activity of the immobilized enzyme was 40 degrees C and the enzyme was stable up to 50 degrees C. The D-Al was active at pH 10.0 and was stable in the range of pH 6.0-11.0. The enzyme required manganese ions for maximum activity. Three immobilized enzymes, D-xylose isomerase (D-XI), D-tagatose 3-epimerase (D-TE and D-AI were used for the preparation of D-arabinose from D-xylose in a coupling reaction. After completion of the reaction, degradation of D-xylulose was carried out by Saccharomyces cerevisiae. The reaction mixture containing D-Xylose, D-ribulose and the product was then separated by ion exchange column chromatography. After crystallization, the product was checked by HPLC, IR spectroscopy, NMR spectroscopy and optical rotation measurements. Finally, 2.0 g of D-arabinose could be obtained from 5 g of the substrate.

The anaerobic linalool metabolism in Thauera linaloolentis 47 Lol.

Science.gov (United States)

Marmulla, Robert; Cala, Edinson Puentes; Markert, Stephanie; Schweder, Thomas; Harder, Jens

2016-04-27

The betaproteobacterium Thauera linaloolentis 47Lol(T) was isolated on the tertiary monoterpene alcohol (R,S)-linalool as sole carbon and energy source under denitrifying conditions. Growth experiments indicated the formation of geraniol and geranial. Thus, a 3,1-hydroxyl-Δ(1)-Δ(2)-mutase (linalool isomerase) activity may initiate the degradation, followed by enzymes of the acyclic terpene utilization (Atu) and leucine/isovalerate utilization (Liu) pathways that were extensively studied in Pseudomonas spp. growing on citronellol or geraniol. A transposon mutagenesis yielded 39 transconjugants that could not grow anaerobically on linalool and nitrate in liquid medium. The deficiencies were apparently based on gene functions required to overcome the toxicity of linalool, but not due to inactivation of genes in the degradation pathway. Growing cultures formed geraniol and geranial transiently, but also geranic acid. Analysis of expressed proteins detected several enzymes of the Atu and Liu pathways. The draft genome of T. linaloolentis 47Lol(T) had atu and liu genes with homology to those of Pseudomonas spp.. The in comparison to monoterpenes larger toxicity of monoterpene alcohols is defeated by several modifications of the cellular structure and metabolism in Thauera linaloolentis 47Lol(T). The acyclic terpene utilization pathway is used in T. linaloolentis 47Lol(T) during growth on (R,S)-linalool and nitrate under anoxic conditions. This is the first experimental verification of an active Atu pathway outside of the genus Pseudomonas.

Anaerobic Digestion Foaming Causes

OpenAIRE

Ganidi, Nafsika

2008-01-01

Anaerobic digestion foaming has been encountered in several sewage treatment plants in the UK. Foaming has raised major concerns for the water utilities due to significant impacts on process efficiency and operational costs. Several foaming causes have been suggested over the past few years by researchers. However, the supporting experimental information is limited and in some cases site specific. The present report aimed to provide a better understanding of the anaerobic di...

75 FR 8920 - Grant of Authority for Subzone Status; Danisco USA, Inc., Sweeteners Division (Xylitol, Xylose...

Science.gov (United States)

2010-02-26

... Status; Danisco USA, Inc., Sweeteners Division (Xylitol, Xylose, Galactose and Mannose); Thomson, IL... subzone at the xylitol, xylose, galactose and mannose manufacturing facility of Danisco USA, Inc... xylitol, xylose, galactose and mannose at the facility of Danisco USA, Inc., Sweeteners Division, located...

Biohydrogen production from xylose at extreme thermophilic temperatures (70 degrees C) by mixed culture fermentation

DEFF Research Database (Denmark)

Kongjan, Prawit; Min, Booki; Angelidaki, Irini

2009-01-01

/L. Addition of yeast extract in the cultivation medium resulted in significant improvement of hydrogen yield. The main metabolic products during xylose fermentation were acetate, ethanol, and lactate. The specific growth rates were able to fit the experimental points relatively well with Haldane equation...... solid wastes at 70 degrees C. The highest hydrogen yield of 1.62 +/- 0.02 mol-H-2/Mol-xylose(consumed) was obtained at initial xylose concentration of 0.5 g/L with synthetic medium amended with I g/L of yeast extract. Lower hydrogen yield was achieved at initial xylose concentration higher than 2 g...

Biodegradation and reversible inhibitory impact of sulfamethoxazole on the utilization of volatile fatty acids during anaerobic treatment of pharmaceutical industry wastewater

International Nuclear Information System (INIS)

Cetecioglu, Zeynep; Ince, Bahar; Gros, Meritxell; Rodriguez-Mozaz, Sara; Barceló, Damia; Ince, Orhan; Orhon, Derin

2015-01-01

This study evaluated the chronic impact and biodegradability of sulfamethoxazole under anaerobic conditions. For this purpose, a lab-scale anaerobic sequencing batch reactor was operated in a sequence of different phases with gradually increasing sulfamethoxazole doses of 1 to 45 mg/L. Conventional parameters, such as COD, VFA, and methane generation, were monitored with corresponding antimicrobial concentrations in the reactor and the methanogenic activity of the sludge. The results revealed that anaerobic treatment was suitable for pharmaceutical industry wastewater with concentrations of up to 40 mg/L of sulfamethoxazole. Higher levels exerted toxic effects on the microbial community under anaerobic conditions, causing the inhibition of substrate/COD utilization and biogas generation and leading to a total collapse of the reactor. The adverse long-term impact was quite variable for fermentative bacteria and methanogenic achaea fractions of the microbial community based on changes inflicted on the composition of the residual organic substrate and mRNA expression of the key enzymes. - Highlights: • Chronic impact of sulfamethoxazole was lethal at 45 mg/L on the microbial community. • Sulfamethoxazole was highly biodegradable under anaerobic conditions. • While the COD removal stopped, the sorption of sulfamethoxazole into the sludge increased. • Sulfamethoxazole has a reversible inhibitory effect on acetoclastic methanogens

Biodegradation and reversible inhibitory impact of sulfamethoxazole on the utilization of volatile fatty acids during anaerobic treatment of pharmaceutical industry wastewater

Energy Technology Data Exchange (ETDEWEB)

Cetecioglu, Zeynep, E-mail: cetecioglu@itu.edu.tr [Istanbul Technical University, Environmental Engineering Department, 34469 Maslak, Istanbul (Turkey); Catalan Institute for Water Research (ICRA), Emili Grahit 101, 17003 Girona (Spain); Ince, Bahar [Bogazici University, Institute of Environmental Sciences, Rumelihisarustu - Bebek, 34342 Istanbul (Turkey); Gros, Meritxell; Rodriguez-Mozaz, Sara; Barceló, Damia [Catalan Institute for Water Research (ICRA), Emili Grahit 101, 17003 Girona (Spain); Ince, Orhan; Orhon, Derin [Istanbul Technical University, Environmental Engineering Department, 34469 Maslak, Istanbul (Turkey)

2015-12-01

This study evaluated the chronic impact and biodegradability of sulfamethoxazole under anaerobic conditions. For this purpose, a lab-scale anaerobic sequencing batch reactor was operated in a sequence of different phases with gradually increasing sulfamethoxazole doses of 1 to 45 mg/L. Conventional parameters, such as COD, VFA, and methane generation, were monitored with corresponding antimicrobial concentrations in the reactor and the methanogenic activity of the sludge. The results revealed that anaerobic treatment was suitable for pharmaceutical industry wastewater with concentrations of up to 40 mg/L of sulfamethoxazole. Higher levels exerted toxic effects on the microbial community under anaerobic conditions, causing the inhibition of substrate/COD utilization and biogas generation and leading to a total collapse of the reactor. The adverse long-term impact was quite variable for fermentative bacteria and methanogenic achaea fractions of the microbial community based on changes inflicted on the composition of the residual organic substrate and mRNA expression of the key enzymes. - Highlights: • Chronic impact of sulfamethoxazole was lethal at 45 mg/L on the microbial community. • Sulfamethoxazole was highly biodegradable under anaerobic conditions. • While the COD removal stopped, the sorption of sulfamethoxazole into the sludge increased. • Sulfamethoxazole has a reversible inhibitory effect on acetoclastic methanogens.

Xylitol production from xylose mother liquor: a novel strategy that combines the use of recombinant Bacillus subtilis and Candida maltosa

Science.gov (United States)

2011-01-01

Background Xylose mother liquor has high concentrations of xylose (35%-40%) as well as other sugars such as L-arabinose (10%-15%), galactose (8%-10%), glucose (8%-10%), and other minor sugars. Due to the complexity of this mother liquor, further isolation of xylose by simple method is not possible. In China, more than 50,000 metric tons of xylose mother liquor was produced in 2009, and the management of sugars like xylose that present in the low-cost liquor is a problem. Results We designed a novel strategy in which Bacillus subtilis and Candida maltosa were combined and used to convert xylose in this mother liquor to xylitol, a product of higher value. First, the xylose mother liquor was detoxified with the yeast C. maltosa to remove furfural and 5-hydromethylfurfural (HMF), which are inhibitors of B. subtilis growth. The glucose present in the mother liquor was also depleted by this yeast, which was an added advantage because glucose causes carbon catabolite repression in B. subtilis. This detoxification treatment resulted in an inhibitor-free mother liquor, and the C. maltosa cells could be reused as biocatalysts at a later stage to reduce xylose to xylitol. In the second step, a recombinant B. subtilis strain with a disrupted xylose isomerase gene was constructed. The detoxified xylose mother liquor was used as the medium for recombinant B. subtilis cultivation, and this led to L-arabinose depletion and xylose enrichment of the medium. In the third step, the xylose was further reduced to xylitol by C. maltosa cells, and crystallized xylitol was obtained from this yeast transformation medium. C. maltosa transformation of the xylose-enriched medium resulted in xylitol with 4.25 g L-1·h-1 volumetric productivity and 0.85 g xylitol/g xylose specific productivity. Conclusion In this study, we developed a biological method for the purification of xylose from xylose mother liquor and subsequent preparation of xylitol by C. maltosa-mediated biohydrogenation of xylose

Xylitol production from xylose mother liquor: a novel strategy that combines the use of recombinant Bacillus subtilis and Candida maltosa

Directory of Open Access Journals (Sweden)

Jiang Mingguo

2011-02-01

Full Text Available Abstract Background Xylose mother liquor has high concentrations of xylose (35%-40% as well as other sugars such as L-arabinose (10%-15%, galactose (8%-10%, glucose (8%-10%, and other minor sugars. Due to the complexity of this mother liquor, further isolation of xylose by simple method is not possible. In China, more than 50,000 metric tons of xylose mother liquor was produced in 2009, and the management of sugars like xylose that present in the low-cost liquor is a problem. Results We designed a novel strategy in which Bacillus subtilis and Candida maltosa were combined and used to convert xylose in this mother liquor to xylitol, a product of higher value. First, the xylose mother liquor was detoxified with the yeast C. maltosa to remove furfural and 5-hydromethylfurfural (HMF, which are inhibitors of B. subtilis growth. The glucose present in the mother liquor was also depleted by this yeast, which was an added advantage because glucose causes carbon catabolite repression in B. subtilis. This detoxification treatment resulted in an inhibitor-free mother liquor, and the C. maltosa cells could be reused as biocatalysts at a later stage to reduce xylose to xylitol. In the second step, a recombinant B. subtilis strain with a disrupted xylose isomerase gene was constructed. The detoxified xylose mother liquor was used as the medium for recombinant B. subtilis cultivation, and this led to L-arabinose depletion and xylose enrichment of the medium. In the third step, the xylose was further reduced to xylitol by C. maltosa cells, and crystallized xylitol was obtained from this yeast transformation medium. C. maltosa transformation of the xylose-enriched medium resulted in xylitol with 4.25 g L-1·h-1 volumetric productivity and 0.85 g xylitol/g xylose specific productivity. Conclusion In this study, we developed a biological method for the purification of xylose from xylose mother liquor and subsequent preparation of xylitol by C. maltosa

Partial oxidation of D-xylose to maleic anhydride and acrylic acid over vanadyl pyrophosphate

International Nuclear Information System (INIS)

Ghaznavi, Touraj; Neagoe, Cristian; Patience, Gregory S.

2014-01-01

Xylose is the second most abundant sugar after glucose. Despite its tremendous potential to serve as a renewable feedstock, few commercial processes exploit this resource. Here, we report a new technology in which a two-fluid nozzle atomizes a xylose-water solution into a capillary fluidized bed operating above 300 °C. Xylose-water droplets form at the tip of the injector, vaporize then react with a heterogeneous mixed oxide catalyst. A syringe pump metered the solution to the reactor charged with 1 g of catalyst. Product yield over vanadyl pyrophosphate was higher compared to molybdenum trioxide-cobalt oxide and iron molybdate; it reached 25% for maleic anhydride, 17% for acrylic acid and 11% for acrolein. Gas residence time was 0.2 s. The catalyst was free of coke even after operating for 4 h – based on a thermogravimetric analysis of catalyst withdrawn from the reactor. Below 300 °C, powder agglomerated at the tip of the injector at 300 °C; it also agglomerated with a xylose mass fraction of 7% in water. - Highlights: • D-xylose reacts to form maleic anhydride and acrylic acid above 250 °C. • Vanadyl pyrophosphate is both active and selective for maleic and acrylic acid. • Acid and acrolein yield approaches 50% for a xylose mass fraction of 3% in water. • Catalyst agglomerates at low temperatures and high xylose aqueous mass fraction. • Atomization quality is a determining factor to minimize agglomeration

Microwave-Assisted Green Production of Furfural from D-xylose of Sugarcane Bagasse

Directory of Open Access Journals (Sweden)

Sílvio Vaz Jr.

2015-10-01

Full Text Available D-xylose is a component of sugarcane bagasse that can be used as a renewable resource for the production of a variety of chemicals. By means of catalytic reactions in an aqueous medium, it was determined that D-xylose can efficiently be converted into furfural by the application of microwave as a green synthetic methodology. The highest yields of furfural were obtained at a HCl concentration of 4 mg/mL. When the reaction was performed at 200 °C, an optimum yield of 64% of furfural was observed after 10 min of reaction time, with 95% of the D-xylose being converted.

Metabolism of Hydrocarbons in n-Alkane-Utilizing Anaerobic Bacteria.

Science.gov (United States)

Wilkes, Heinz; Buckel, Wolfgang; Golding, Bernard T; Rabus, Ralf

2016-01-01

The glycyl radical enzyme-catalyzed addition of n-alkanes to fumarate creates a C-C-bond between two concomitantly formed stereogenic carbon centers. The configurations of the two diastereoisomers of the product resulting from n-hexane activation by the n-alkane-utilizing denitrifying bacterium strain HxN1, i.e. (1-methylpentyl)succinate, were assigned as (2S,1'R) and (2R,1'R). Experiments with stereospecifically deuterated n-(2,5-2H2)hexanes revealed that exclusively the pro-S hydrogen atom is abstracted from C2 of the n-alkane by the enzyme and later transferred back to C3 of the alkylsuccinate formed. These results indicate that the alkylsuccinate-forming reaction proceeds with an inversion of configuration at the carbon atom (C2) of the n-alkane forming the new C-C-bond, and thus stereochemically resembles a SN2-type reaction. Therefore, the reaction may occur in a concerted manner, which may avoid the highly energetic hex-2-yl radical as an intermediate. The reaction is associated with a significant primary kinetic isotope effect (kH/kD ≥3) for hydrogen, indicating that the homolytic C-H-bond cleavage is involved in the first irreversible step of the reaction mechanism. The (1-methylalkyl)succinate synthases of n-alkane-utilizing anaerobic bacteria apparently have very broad substrate ranges enabling them to activate not only aliphatic but also alkyl-aromatic hydrocarbons. Thus, two denitrifiers and one sulfate reducer were shown to convert the nongrowth substrate toluene to benzylsuccinate and further to the dead-end product benzoyl-CoA. For this purpose, however, the modified β-oxidation pathway known from alkylbenzene-utilizing bacteria was not employed, but rather the pathway used for n-alkane degradation involving CoA ligation, carbon skeleton rearrangement and decarboxylation. Furthermore, various n-alkane- and alkylbenzene-utilizing denitrifiers and sulfate reducers were found to be capable of forming benzyl alcohols from diverse alkylbenzenes

Lactic acid production from xylose by Geobacillus stearothermophilus strain 15

Science.gov (United States)

Kunasundari, B.; Naresh, S.; Chu, J. E.

2017-09-01

Lactic acid is an important compound with a wide range of industrial applications. The present study tested the efficiency of xylose, as a sole carbon source to be converted to lactic acid by Geobacillus stearothermophilus strain 15. To the best of our knowledge, limited information is available on the directed fermentation of xylose to lactic acid by this bacterium. The effects of different parameters such as temperature, pH, incubation time, agitation speed, concentrations of nitrogen and carbon sources on the lactic acid production were investigated statistically. It was found that the bacterium exhibited poor assimilation of xylose to lactic acid. Temperature, agitation rate and incubation time were determined to improve the lactic acid production slightly. The highest lactic acid yield obtained was 8.9% at 45°C, 300 RPM, 96 h, pH of 6.0 with carbon and nitrogen source concentrations were fixed at 5% w/v.

L-Lactic acid production from glucose and xylose with engineered strains of Saccharomyces cerevisiae: aeration and carbon source influence yields and productivities.

Science.gov (United States)

Novy, Vera; Brunner, Bernd; Nidetzky, Bernd

2018-04-11

Saccharomyces cerevisiae, engineered for L-lactic acid production from glucose and xylose, is a promising production host for lignocellulose-to-lactic acid processes. However, the two principal engineering strategies-pyruvate-to-lactic acid conversion with and without disruption of the competing pyruvate-to-ethanol pathway-have not yet resulted in strains that combine high lactic acid yields (Y LA ) and productivities (Q LA ) on both sugar substrates. Limitations seemingly arise from a dependency on the carbon source and the aeration conditions, but the underlying effects are poorly understood. We have recently presented two xylose-to-lactic acid converting strains, IBB14LA1 and IBB14LA1_5, which have the L-lactic acid dehydrogenase from Plasmodium falciparum (pfLDH) integrated at the pdc1 (pyruvate decarboxylase) locus. IBB14LA1_5 additionally has its pdc5 gene knocked out. In this study, the influence of carbon source and oxygen on Y LA and Q LA in IBB14LA1 and IBB14LA1_5 was investigated. In anaerobic fermentation IBB14LA1 showed a higher Y LA on xylose (0.27 g g Xyl -1 ) than on glucose (0.18 g g Glc -1 ). The ethanol yields (Y EtOH , 0.15 g g Xyl -1 and 0.32 g g Glc -1 ) followed an opposite trend. In IBB14LA1_5, the effect of the carbon source on Y LA was less pronounced (~ 0.80 g g Xyl -1 , and 0.67 g g Glc -1 ). Supply of oxygen accelerated glucose conversions significantly in IBB14LA1 (Q LA from 0.38 to 0.81 g L -1  h -1 ) and IBB14LA1_5 (Q LA from 0.05 to 1.77 g L -1  h -1 ) at constant Y LA (IBB14LA1 ~ 0.18 g g Glc -1 ; IBB14LA1_5 ~ 0.68 g g Glc -1 ). In aerobic xylose conversions, however, lactic acid production ceased completely in IBB14LA1 and decreased drastically in IBB14LA1_5 (Y LA aerobic ≤ 0.25 g g Xyl -1 and anaerobic ~ 0.80 g g Xyl -1 ) at similar Q LA (~ 0.04 g L -1  h -1 ). Switching from aerobic to microaerophilic conditions (pO 2  ~ 2%) prevented lactic acid metabolization, observed for

Glucose (xylose) isomerase production from thermotolerant and ...

African Journals Online (AJOL)

Owner

2012-11-13

Nov 13, 2012 ... in the production of the high fructose corn syrup (HFCS) from corn starch. ... Key words: Glucose isomerase, xylose isomerase, enzyme activity, Klebsiella, ... Soil, water, and manure (five samples each) were collected from.

Statistical optimization of fermentative hydrogen production from xylose by newly isolated Enterobacter sp. CN1

Energy Technology Data Exchange (ETDEWEB)

Long, Chuannan; Cui, Jingjing; Liu, Zuotao; Liu, Yuntao; Hu, Zhong [Department of Biology, Shantou University, Shantou 515063 (China); Long, Minnan [The School of Energy Research, Xiamen University, Xiamen 361005 (China)

2010-07-15

Statistical experimental designs were applied for the optimization of medium constituents for hydrogen production from xylose by newly isolated Enterobacter sp. CN1. Using Plackett-Burman design, xylose, FeSO{sub 4} and peptone were identified as significant variables which highly influenced hydrogen production. The path of steepest ascent was undertaken to approach the optimal region of the three significant factors. These variables were subsequently optimized using Box-Behnken design of response surface methodology (RSM). The optimum conditions were found to be xylose 16.15 g/L, FeSO{sub 4} 250.17 mg/L, peptone 2.54 g/L. Hydrogen production at these optimum conditions was 1149.9 {+-} 65 ml H{sub 2}/L medium. Under different carbon sources condition, the cumulative hydrogen volume were 1217 ml H{sub 2}/L xylose medium, 1102 ml H{sub 2}/L glucose medium and 977 ml H{sub 2}/L sucrose medium; the maximum hydrogen yield were 2.0 {+-} 0.05 mol H{sub 2}/mol xylose, 0.64 mol H{sub 2}/mol glucose. Fermentative hydrogen production from xylose by Enterobacter sp. CN1 was superior to glucose and sucrose. (author)

Rational and evolutionary engineering approaches uncover a small set of genetic changes efficient for rapid xylose fermentation in Saccharomyces cerevisiae.

Directory of Open Access Journals (Sweden)

Soo Rin Kim

Full Text Available Economic bioconversion of plant cell wall hydrolysates into fuels and chemicals has been hampered mainly due to the inability of microorganisms to efficiently co-ferment pentose and hexose sugars, especially glucose and xylose, which are the most abundant sugars in cellulosic hydrolysates. Saccharomyces cerevisiae cannot metabolize xylose due to a lack of xylose-metabolizing enzymes. We developed a rapid and efficient xylose-fermenting S. cerevisiae through rational and inverse metabolic engineering strategies, comprising the optimization of a heterologous xylose-assimilating pathway and evolutionary engineering. Strong and balanced expression levels of the XYL1, XYL2, and XYL3 genes constituting the xylose-assimilating pathway increased ethanol yields and the xylose consumption rates from a mixture of glucose and xylose with little xylitol accumulation. The engineered strain, however, still exhibited a long lag time when metabolizing xylose above 10 g/l as a sole carbon source, defined here as xylose toxicity. Through serial-subcultures on xylose, we isolated evolved strains which exhibited a shorter lag time and improved xylose-fermenting capabilities than the parental strain. Genome sequencing of the evolved strains revealed that mutations in PHO13 causing loss of the Pho13p function are associated with the improved phenotypes of the evolved strains. Crude extracts of a PHO13-overexpressing strain showed a higher phosphatase activity on xylulose-5-phosphate (X-5-P, suggesting that the dephosphorylation of X-5-P by Pho13p might generate a futile cycle with xylulokinase overexpression. While xylose consumption rates by the evolved strains improved substantially as compared to the parental strain, xylose metabolism was interrupted by accumulated acetate. Deletion of ALD6 coding for acetaldehyde dehydrogenase not only prevented acetate accumulation, but also enabled complete and efficient fermentation of xylose as well as a mixture of glucose and

Production of furfural from rice straw by microbial treatment. (II). Production of furfural from xylose by acid treatment

Energy Technology Data Exchange (ETDEWEB)

Kim, W.S.; Yoo, I.S.; Kang, S.K.

1984-01-01

The reaction conditions and mechanism of furfural production from xylose by acid treatment were studied. The xylose was obtained from rice straw. Furfural yield at batch-isothermal conditions was a function of initial xylose concentration H2SO4 concentration, reaction temperature and reaction time. And when the initial xylose concentration was low, the results were consistent with those of Root's reaction mechanism. Maximum furfural yield was obtained under conditions of H2SO4 concentration 0.2N, initial xylose concentration 0.0067 M, temperature 200 degrees, and reaction time 10 min.

Degradative capacities and bioaugmentation potential of an anaerobic benzene-degrading bacterium strain DN11

Energy Technology Data Exchange (ETDEWEB)

Yuki Kasai; Yumiko Kodama; Yoh Takahata; Toshihiro Hoaki; Kazuya Watanabe [Marine Biotechnology Institute, Kamaishi (Japan)

2007-09-15

Azoarcus sp. strain DN11 is a denitrifying bacterium capable of benzene degradation under anaerobic conditions. The present study evaluated strain DN11 for its application to bioaugmentation of benzene-contaminated underground aquifers. Strain DN11 could grow on benzene, toluene, m-xylene, and benzoate as the sole carbon and energy sources under nitrate-reducing conditions, although o- and p-xylenes were transformed in the presence of toluene. Phenol was not utilized under anaerobic conditions. Kinetic analysis of anaerobic benzene degradation estimated its apparent affinity and inhibition constants to be 0.82 and 11 {mu}M, respectively. Benzene-contaminated groundwater taken from a former coal-distillation plant site in Aichi, Japan was anaerobically incubated in laboratory bottles and supplemented with either inorganic nutrients (nitrogen, phosphorus, and nitrate) alone, or the nutrients plus strain DN11, showing that benzene was significantly degraded only when DN11 was introduced. Denaturing gradient gel electrophoresis of PCR-amplified 16S rRNA gene fragments, and quantitative PCR revealed that DN11 decreased after benzene was degraded. Following the decrease in DN11 16S rRNA gene fragments corresponding to bacteria related to Owenweeksia hongkongensis and Pelotomaculum isophthalicum, appeared as strong bands, suggesting possible metabolic interactions in anaerobic benzene degradation. Results suggest that DN11 is potentially useful for degrading benzene that contaminates underground aquifers at relatively low concentrations. 50 refs., 6 figs., 1 tab.

Enhanced Furfural Yields from Xylose Dehydration in the gamma-Valerolactone/Water Solvent System at Elevated Temperatures.

Science.gov (United States)

Sener, Canan; Motagamwala, Ali Hussain; Alonso, David Martin; Dumesic, James

2018-05-18

High yields of furfural (>90%) were achieved from xylose dehydration in a sustainable solvent system composed of -valerolactone (GVL), a biomass derived solvent, and water. It is identified that high reaction temperatures (e.g., 498 K) are required to achieve high furfural yield. Additionally, it is shown that the furfural yield at these temperatures is independent of the initial xylose concentration, and high furfural yield is obtained for industrially relevant xylose concentrations (10 wt%). A reaction kinetics model is developed to describe the experimental data obtained with solvent system composed of 80 wt% GVL and 20 wt% water across the range of reaction conditions studied (473 - 523 K, 1-10 mM acid catalyst, 66 - 660 mM xylose concentration). The kinetic model demonstrates that furfural loss due to bimolecular condensation of xylose and furfural is minimized at elevated temperature, whereas carbon loss due to xylose degradation increases with increasing temperature. Accordingly, the optimal temperature range for xylose dehydration to furfural in the GVL/H2O solvent system is identified to be from 480 to 500 K. Under these reaction conditions, furfural yield of 93% is achieved at 97% xylan conversion from lignocellulosic biomass (maple wood). © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Changes in kenaf properties and chemistry as a function of growing time

Science.gov (United States)

Roger M. Rowell; James S. Han

1999-01-01

Kenaf Tainung 1 cultivar was grown in Madison, WI in 1994. The ratio of core to bast fiber, total plant yield, protein, ash, fiber length, extractives, lignin, and sugar content were determined as a function of growing age. Ash, protein, extractives, L-arabinose, L-rhamnose, D-galactose, and D-mannose contents decreased while lignin, D-glucose and D-xylose content...

Identification of anaerobic microorganisms for converting kitchen waste to biogas

International Nuclear Information System (INIS)

Amirhossein Malakahmad; Shahrom Mohd Zain; Noor Ezlin Ahmad Basri; Shamsul Rahman Mohamed Kutty; Mohd Hasnain Isa

2010-01-01

Anaerobic digestion process is one of the alternative methods to convert organic waste into methane gas which is a fuel and energy source. Activities of various kinds of microorganisms are the main factor for anaerobic digestion which produces methane gas. Therefore, in this study a modified Anaerobic Baffled Reactor (ABR) with working volume of 50 liters was designed to identify the microorganisms through biogas production. The mixture of 75% kitchen waste and 25% sewage sludge was used as substrate. Observations on microorganisms in the ABR showed that there exists a small amount of protozoa (5%) and fungi (2%) in the system, but almost 93% of the microorganism population consists of bacteria. It is definitely clear that bacteria are responsible for anaerobic biodegradation of kitchen waste. Results show that in the acidification zone of the ABR (front compartments of reactor) fast growing bacteria capable of growth at high substrate levels and reduced pH was dominant. A shift to slower growing scavenging bacteria that grow better at higher pH was occurring towards the end of the reactor. Due to the ability of activity in acetate environment the percentages of Methanococcus, Methanosarcina and Methanotrix were higher than other kinds of methane former in the system. (Author)

The anaerobic digestion process

Energy Technology Data Exchange (ETDEWEB)

Rivard, C.J. [National Renewable Energy Lab., Golden, CO (United States); Boone, D.R. [Oregon Graduate Inst., Portland, OR (United States)

1996-01-01

The microbial process of converting organic matter into methane and carbon dioxide is so complex that anaerobic digesters have long been treated as {open_quotes}black boxes.{close_quotes} Research into this process during the past few decades has gradually unraveled this complexity, but many questions remain. The major biochemical reactions for forming methane by methanogens are largely understood, and evolutionary studies indicate that these microbes are as different from bacteria as they are from plants and animals. In anaerobic digesters, methanogens are at the terminus of a metabolic web, in which the reactions of myriads of other microbes produce a very limited range of compounds - mainly acetate, hydrogen, and formate - on which the methanogens grow and from which they form methane. {open_quotes}Interspecies hydrogen-transfer{close_quotes} and {open_quotes}interspecies formate-transfer{close_quotes} are major mechanisms by which methanogens obtain their substrates and by which volatile fatty acids are degraded. Present understanding of these reactions and other complex interactions among the bacteria involved in anaerobic digestion is only now to the point where anaerobic digesters need no longer be treated as black boxes.

Isolation and characterization of two novel ethanol-tolerant facultative-anaerobic thermophilic bacteria strains from waste compost.

Science.gov (United States)

Fong, Jiunn C N; Svenson, Charles J; Nakasugi, Kenlee; Leong, Caine T C; Bowman, John P; Chen, Betty; Glenn, Dianne R; Neilan, Brett A; Rogers, Peter L

2006-10-01

In a search for potential ethanologens, waste compost was screened for ethanol-tolerant thermophilic microorganisms. Two thermophilic bacterial strains, M5EXG and M10EXG, with tolerance of 5 and 10% (v/v) ethanol, respectively, were isolated. Both isolates are facultative anaerobic, non-spore forming, non-motile, catalase-positive, oxidase-negative, Gram-negative rods that are capable of utilizing a range of carbon sources including arabinose, galactose, mannose, glucose and xylose and produce low amounts of ethanol, acetate and lactate. Growth of both isolates was observed in fully defined minimal media within the temperature range 50-80 degrees C and pH 6.0-8.0. Phylogenetic analysis of the 16S rDNA sequences revealed that both isolates clustered with members of subgroup 5 of the genus Bacillus. G+C contents and DNA-DNA relatedness of M5EXG and M10EXG revealed that they are strains belonging to Geobacillus thermoglucosidasius. However, physiological and biochemical differences were evident when isolates M5EXG and M10EXG were compared with G. thermoglucosidasius type strain (DSM 2542(T)). The new thermophilic, ethanol-tolerant strains of G. thermoglucosidasius may be candidates for ethanol production at elevated temperatures.

Analytical Validation of a New Enzymatic and Automatable Method for d-Xylose Measurement in Human Urine Samples

Directory of Open Access Journals (Sweden)

Israel Sánchez-Moreno

2017-01-01

Full Text Available Hypolactasia, or intestinal lactase deficiency, affects more than half of the world population. Currently, xylose quantification in urine after gaxilose oral administration for the noninvasive diagnosis of hypolactasia is performed with the hand-operated nonautomatable phloroglucinol reaction. This work demonstrates that a new enzymatic xylose quantification method, based on the activity of xylose dehydrogenase from Caulobacter crescentus, represents an excellent alternative to the manual phloroglucinol reaction. The new method is automatable and facilitates the use of the gaxilose test for hypolactasia diagnosis in the clinical practice. The analytical validation of the new technique was performed in three different autoanalyzers, using buffer or urine samples spiked with different xylose concentrations. For the comparison between the phloroglucinol and the enzymatic assays, 224 urine samples of patients to whom the gaxilose test had been prescribed were assayed by both methods. A mean bias of −16.08 mg of xylose was observed when comparing the results obtained by both techniques. After adjusting the cut-off of the enzymatic method to 19.18 mg of xylose, the Kappa coefficient was found to be 0.9531, indicating an excellent level of agreement between both analytical procedures. This new assay represents the first automatable enzymatic technique validated for xylose quantification in urine.

Genes for Uranium Bioremediation in the Anaerobic Sulfate-Reducing Bacteria: Desulfovibrio mutants with altered sensitivity to oxidative stress

International Nuclear Information System (INIS)

Payne, Rayford B.; Ringbauer, Joseph A. Jr.; Wall, Judy D.

2006-01-01

Sulfate-reducing bacteria of the genus Desulfovibrio are ubiquitous in anaerobic environments such as groundwater, sediments, and the gastrointestinal tract of animals. Because of the ability of Desulfovibrio to reduce radionuclides and metals through both enzymatic and chemical means, they have been proposed as a means to bioremediate heavy metal contaminated sites. Although classically thought of as strict anaerobes, Desulfovibrio species are surprisingly aerotolerant. Our objective is to understand the response of Desulfovibrio to oxidative stress so that we may more effectively utilize them in bioremediation of heavy metals in mixed aerobic-anaerobic environments. The enzymes superoxide dismutase, superoxide reductase, catalase, and rubrerythrin have been shown by others to be involved in the detoxification of reactive oxygen species in Desulfovibrio. Some members of the genus Desulfovibrio can even reduce molecular oxygen to water via a membrane bound electron transport chain with the concomitant production of ATP, although their ability to grow with oxygen as the sole electron acceptor is still questioned.

Continuous xylose fermentation by Clostridium acetobutylicum – Kinetics and energetics issues under acidogenesis conditions

NARCIS (Netherlands)

Procentese, A.; Raganati, F.; Olivieri, G.; Russo, M.E.; Salatino, P.; Marzocchella, A.

2014-01-01

The paper reports the assessment of the growth kinetics of Clostridium acetobutylicum DSM 792 adopting xylose as carbon source. Xylose is the fundamental component of hemicellulose hydrolysis, a relevant fraction of lignocellulosic feedstocks for biofuel production. Tests were carried out in a CSTR

A synthetic hybrid promoter for xylose-regulated control of gene expression in Saccharomyces yeasts

Science.gov (United States)

Metabolism of non-glucose carbon sources is often highly regulated at the transcriptional and post-translational levels. This level of regulation is lacking in Saccharomyces cerevisiae strains engineered to metabolize xylose. To better control transcription in S. cerevisiae, the xylose-dependent, DN...

Isolation and Characterization of Acetate-Utilizing Anaerobes from a Freshwater Sediment.

Science.gov (United States)

Scholten, J.C.M.; Stams, A.J.M.

2000-12-01

Acetate-degrading anaerobic microorganisms in freshwater sediment were quantified by the most probable number technique. From the highest dilutions a methanogenic, a sulfate-reducing, and a nitrate-reducing microorganism were isolated with acetate as substrate. The methanogen (culture AMPB-Zg) was non-motile and rod-shaped with blunted ends (0.5-1 mm x 3-4 mm long). Doubling times with acetate at 30-35 degrees C were 5.6-8.1 days. The methanogen grew only on acetate. Analysis of the 16S rRNA sequence showed that AMPB-Zg is closely related to Methanosaeta concilii. The isolated sulfate-reducing bacterium (strain ASRB-Zg) was rod-shaped with pointed ends (0.5-0.7 mm x 1.5-3.5 mm long), weakly motile, spore forming, and gram positive. At the optimum growth temperature of 30 degrees C the doubling times with acetate were 3.9-5.3 days. The bacterium grew on a range of organic acids, such as acetate, butyrate, fumarate, and benzoate, but did not grow autotrophically with H2, CO2, and sulfate. The closest relative of strain ASRB-Zg is Desulfotomaculum acetoxidans. The nitrate-reducing bacterium (strain ANRB-Zg) was rod-shaped (0.5-0.7 mm x 0.7-1 mm long), weakly motile, and gram negative. Optimum growth with acetate occurred at 20-25 degrees C. The bacterium grew on a range of organic substrates, such as acetate, butyrate, lactate, and glucose, and did grow autotrophically with H2, CO2, and oxygen but not with nitrate. In the presence of acetate and nitrate, thiosulfate was oxidized to sulfate. Phylogenetically, the closest relative of strain ANRB-Zg is Variovorax paradoxus.

New Protocol Based on UHPLC-MS/MS for Quantitation of Metabolites in Xylose-Fermenting Yeasts

Science.gov (United States)

Campos, Christiane Gonçalves; Veras, Henrique César Teixeira; de Aquino Ribeiro, José Antônio; Costa, Patrícia Pinto Kalil Gonçalves; Araújo, Katiúscia Pereira; Rodrigues, Clenilson Martins; de Almeida, João Ricardo Moreira; Abdelnur, Patrícia Verardi

2017-12-01

Xylose fermentation is a bottleneck in second-generation ethanol production. As such, a comprehensive understanding of xylose metabolism in naturally xylose-fermenting yeasts is essential for prospection and construction of recombinant yeast strains. The objective of the current study was to establish a reliable metabolomics protocol for quantification of key metabolites of xylose catabolism pathways in yeast, and to apply this protocol to Spathaspora arborariae. Ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) was used to quantify metabolites, and afterwards, sample preparation was optimized to examine yeast intracellular metabolites. S. arborariae was cultivated using xylose as a carbon source under aerobic and oxygen-limited conditions. Ion pair chromatography (IPC) and hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) were shown to efficiently quantify 14 and 5 metabolites, respectively, in a more rapid chromatographic protocol than previously described. Thirteen and eleven metabolites were quantified in S. arborariae under aerobic and oxygen-limited conditions, respectively. This targeted metabolomics protocol is shown here to quantify a total of 19 metabolites, including sugars, phosphates, coenzymes, monosaccharides, and alcohols, from xylose catabolism pathways (glycolysis, pentose phosphate pathway, and tricarboxylic acid cycle) in yeast. Furthermore, to our knowledge, this is the first time that intracellular metabolites have been quantified in S. arborariae after xylose consumption. The results indicated that fine control of oxygen levels during fermentation is necessary to optimize ethanol production by S. arborariae. The protocol presented here may be applied to other yeast species and could support yeast genetic engineering to improve second generation ethanol production. [Figure not available: see fulltext.

Treatment of industrial wastewaters by anaerobic membrane bioreactors : Implications of substrate characteristics

NARCIS (Netherlands)

Dereli, R.K.

2015-01-01

The success of anaerobic digestion relies on the presence of highly active methanogenic biomass, requiring effective retention of slow growing anaerobic microorganisms inside bioreactor by decoupling the hydraulic retention time (HRT) from solids residence time (SRT) or the employment of long SRTs

Efficient fermentation of xylose to ethanol at high formic acid concentrations by metabolically engineered Saccharomyces cerevisiae

Energy Technology Data Exchange (ETDEWEB)

Hasunuma, Tomohisa; Yoshimura, Kazuya; Matsuda, Fumio [Kobe Univ., Hyogo (Japan). Organization of Advanced Science and Technology; Sung, Kyung-mo; Sanda, Tomoya; Kondo, Akihiko [Kobe Univ., Hyogo (Japan). Dept. of Chemical Science and Engineering

2011-05-15

Recombinant yeast strains highly tolerant to formic acid during xylose fermentation were constructed. Microarray analysis of xylose-fermenting Saccharomyces cerevisiae strain overexpressing endogenous xylulokinase in addition to xylose reductase and xylitol dehydrogenase from Pichia stipitis revealed that upregulation of formate dehydrogenase genes (FDH1 and FDH2) was one of the most prominent transcriptional events against excess formic acid. The quantification of formic acid in medium indicated that the innate activity of FDH was too weak to detoxify formic acid. To reinforce the capability for formic acid breakdown, the FDH1 gene was additionally overexpressed in the xylose-metabolizing recombinant yeast. This modification allowed the yeast to rapidly decompose excess formic acid. The yield and final ethanol concentration in the presence of 20 mM formic acid is as essentially same as that of control. The fermentation profile also indicated that the production of xylitol and glycerol, major by-products in xylose fermentation, was not affected by the upregulation of FDH activity. (orig.)

Potential of xylose-fermented yeast isolated from sugarcane bagasse waste for xylitol production using hydrolysate as carbon source

Directory of Open Access Journals (Sweden)

Kusumawadee Thancharoen

2016-10-01

Full Text Available Xylitol is a high value sugar alcohol that is used as a sweetener. In the past years, the biological process of D-xylose from lignocellulosic material into xylitol has gained increasing interest as an alternative production method. In this study, sugarcane bagasse was used as raw material for xylitol production because of its high efficiency, reduced industrial cost, and high concentration of xylose. Pre-treatment of sugarcane bagasse with sulfuric acid was performed with various conditions. The results showed that the optimum condition was exhibited for 3.1% sulfuric acid at 126°C for 18 min producing 19 g/l xylose. Isolated yeasts from the sugarcane bagasse were selected and tested for xylitol ability from xylose. Results showed that Candida tropicalis KS 10-3 (from 72 isolates had the highest ability and produced 0.47 g xylitol/ g xylose in 96 hrs of cultivation containing 32.30 g/l xylose was used as the production medium.

Effects of lignin-derived phenolic compounds on xylitol production and key enzyme activities by a xylose utilizing yeast Candida athensensis SB18.

Science.gov (United States)

Zhang, Jinming; Geng, Anli; Yao, Chuanyi; Lu, Yinghua; Li, Qingbiao

2012-10-01

Candida athensensis SB18 is potential xylitol producing yeast isolated in Singapore. It has excellent xylose tolerance and is able to produce xylitol in high titer and yield. However, by-products, such as phenolic compounds, derived in lignocellulosic biomass hydrolysate might negatively influence the performance of this strain for xylitol production. In this work, four potential phenolic inhibitors, such as vanillin, syringaldehyde, 4-hydroxybenzaldehyde and phenol, were evaluated for their inhibitory effects on xylitol production by C. athensensis SB18. Phenol was shown to be the most toxic molecule on this microorganism followed by syringaldehyde. Vanillin and 4-hydroxylbenzaldehyde was less toxic than phenol and syringaldehyde, with vanillin being the least toxic. Inhibition was insignificant when the total content of inhibitors was below 1.0 g/L. The presence of phenolic compounds affected the activity of xylose reductase, however not on that of xylitol dehydrogenase. C. athensensis SB18 is therefore a potential xylitol producer from hemicellulosic hydrolysate due to its assimilation of such phenolic inhibitors. Copyright © 2012 Elsevier Ltd. All rights reserved.

Ethanol fermentation from lignocellulosic hydrolysate by a recombinant xylose- and cellooligosaccharide-assimilating yeast strain

Energy Technology Data Exchange (ETDEWEB)

Katahira, Satoshi; Fukuda, Hideki [Kobe Univ. (Japan). Div. of Molecular Science; Mizuike, Atsuko; Kondo, Akihiko [Kobe Univ. (Japan). Dept. of Chemical Science and Engineering

2006-10-15

The sulfuric acid hydrolysate of lignocellulosic biomass, such as wood chips, from the forest industry is an important material for fuel bioethanol production. In this study, we constructed a recombinant yeast strain that can ferment xylose and cellooligosaccharides by integrating genes for the intercellular expressions of xylose reductase and xylitol dehydrogenase from Pichia stipitis, and xylulokinase from Saccharomyces cerevisiae and a gene for displaying ss-glucosidase from Aspergillus acleatus on the cell surface. In the fermentation of the sulfuric acid hydrolysate of wood chips, xylose and cellooligosaccharides were completely fermented after 36 h by the recombinant strain, and then about 30 g/l ethanol was produced from 73 g/l total sugar added at the beginning. In this case, the ethanol yield of this recombinant yeast was much higher than that of the control yeast. These results demonstrate that the fermentation of the lignocellulose hydrolysate is performed efficiently by the recombinant Saccharomyces strain with abilities for xylose assimilation and cellooligosaccharide degradation. (orig.)

Glucose(xylose isomerase production by Streptomyces sp. CH7 grown on agricultural residues

Directory of Open Access Journals (Sweden)

Kankiya Chanitnun

2012-09-01

Full Text Available Streptomyces sp. CH7 was found to efficiently produce glucose(xylose isomerase when grown on either xylan or agricultural residues. This strain produced a glucose(xylose isomerase activity of roughly 1.8 U/mg of protein when it was grown in medium containing 1% xylose as a carbon source. Maximal enzymatic activities of about 5 and 3 U/mg were obtained when 1% xylan and 2.5% corn husks were used, respectively. The enzyme was purified from a mycelial extract to 16-fold purity with only two consecutive column chromatography steps using Macro-prep DEAE and Sephacryl-300, respectively. The approximate molecular weight of the purified enzyme is 170 kDa, and it has four identical subunits of 43.6 kDa as estimated by SDS-PAGE. Its Km values for glucose and xylose were found to be 258.96 and 82.77 mM, respectively, and its Vmax values are 32.42 and 63.64 μM/min/mg, respectively. The purified enzyme is optimally active at 85ºC and pH 7.0. It is stable at pH 5.5-8.5 and at temperatures up to 60ºC after 30 min. These findings indicate that glucose(xylose isomerase from Streptomyces sp. CH7 has the potential for industrial applications, especially for high-fructose syrup production and bioethanol fermentation from hemicellulosic hydrolysates by Saccharomyces cerevisiae.

Enhancing ethanol yields through d-xylose and l-arabinose co-fermentation after construction of a novel high efficient l-arabinose-fermenting Saccharomyces cerevisiae strain.

Science.gov (United States)

Caballero, Antonio; Ramos, Juan Luis

2017-04-01

Lignocellulose contains two pentose sugars, l-arabinose and d-xylose, neither of which is naturally fermented by first generation (1G) ethanol-producing Saccharomyces cerevisiae yeast. Since these sugars are inaccessible to 1G yeast, a significant percentage of the total carbon in bioethanol production from plant residues, which are used in second generation (2G) ethanol production, remains unused. Recombinant Saccharomyces cerevisiae strains capable of fermenting d-xylose are available on the market; however, there are few examples of l-arabinose-fermenting yeasts, and commercially, there are no strains capable of fermenting both d-xylose and l-arabinose because of metabolic incompatibilities when both metabolic pathways are expressed in the same cell. To attempt to solve this problem we have tested d-xylose and l-arabinose co-fermentation. To find efficient alternative l-arabinose utilization pathways to the few existing ones, we have used stringent methodology to screen for new genes (metabolic and transporter functions) to facilitate l-arabinose fermentation in recombinant yeast. We demonstrate the feasibility of this approach in a successfully constructed yeast strain capable of using l-arabinose as the sole carbon source and capable of fully transforming it to ethanol, reaching the maximum theoretical fermentation yield (0.43 g g-1). We demonstrate that efficient co-fermentation of d-xylose and l-arabinose is feasible using two different co-cultured strains, and observed no fermentation delays, yield drops or accumulation of undesired byproducts. In this study we have identified a technically efficient strategy to enhance ethanol yields by 10 % in 2G plants in a process based on C5 sugar co-fermentation.

Anaerobic/oxic/anoxic granular sludge process as an effective nutrient removal process utilizing denitrifying polyphosphate-accumulating organisms.

Science.gov (United States)

Kishida, Naohiro; Kim, Juhyun; Tsuneda, Satoshi; Sudo, Ryuichi

2006-07-01

In a biological nutrient removal (BNR) process, the utilization of denitrifying polyphosphate-accumulating organisms (DNPAOs) has many advantages such as effective use of organic carbon substrates and low sludge production. As a suitable process for the utilization of DNPAOs in BNR, an anaerobic/oxic/anoxic granular sludge (AOAGS) process was proposed in this study. In spite of performing aeration for nitrifying bacteria, the AOAGS process can create anaerobic/anoxic conditions suitable for the cultivation of DNPAOs because anoxic zones exist inside the granular sludge in the oxic phase. Thus, DNPAOs can coexist with nitrifying bacteria in a single reactor. In addition, the usability of DNPAOs in the reactor can be improved by adding the anoxic phase after the oxic phase. These characteristics enable the AOAGS process to attain effective removal of both nitrogen and phosphorus. When acetate-based synthetic wastewater (COD: 600 mg/L, NH4-N: 60 mg/L, PO(4)-P: 10 mg/L) was supplied to a laboratory-scale sequencing batch reactor under the operation of anaerobic/oxic/anoxic cycles, granular sludge with a diameter of 500 microm was successfully formed within 1 month. Although the removal of both nitrogen and phosphorus was almost complete at the end of the oxic phase, a short anoxic period subsequent to the oxic phase was necessary for further removal of nitrogen and phosphorus. As a result, effluent concentrations of NH(4)-N, NO(x)-N and PO(4)-P were always lower than 1 mg/L. It was found that penetration depth of oxygen inside the granular sludge was approximately 100 microm by microsensor measurements. In addition, from the microbiological analysis by fluorescence in situ hybridization, existence depth of polyphosphate-accumulating organisms was further than the maximum oxygen penetration depth. The water quality data, oxygen profiles and microbial community structure demonstrated that DNPAOs inside the granular sludge may be responsible for denitrification in the

Ability for anaerobic growth is not sufficient for development of the petite phenotype in Saccharomyces kluyveri

DEFF Research Database (Denmark)

Møller, Kasper; Olsson, Lisbeth; Piskur, Jure

2001-01-01

Saccharomyces cerevisiae is a petite-phenotype-positive ("petite-positive") yeast, which can successfully grow in the absence of oxygen. On the other hand, Kluyveromyces lactis as well as many other yeasts are petite negative and cannot grow anaerobically. In this paper, we show that Saccharomyces...... kluyveri can grow under anaerobic conditions, but while it can generate respiration-deficient mutants, it cannot generate true petite mutants. From a phylogenetic point of view, S. kluyveri is apparently more closely related to S. cerevisiae than to K. lactis. These observations suggest that the progenitor...... of the modern Saccharomyces and Kluyveromyces yeasts, as well as other related genera, was a petite-negative and aerobic yeast. Upon separation of the K. lactis and S. kluyveri-S. cerevisiae lineages, the latter developed the ability to grow anaerobically. However, while the S. kluyveri lineage has remained...

Use of agricultural by-products for the production of xylitol. I. The production of xylose

Energy Technology Data Exchange (ETDEWEB)

De Menezes, H C

1976-01-01

A Rhizopus species capable of converting xylan into xylose was isolated from the soil, and purified. The xylanase produced by this fungus was capable of producing xylose from corn cob, wheat bran, and rice hulls without prior extraction of the xylan.

Improving L-arabinose utilization of pentose fermenting Saccharomyces cerevisiae cells by heterologous expression of L-arabinose transporting sugar transporters

Directory of Open Access Journals (Sweden)

Boles Eckhard

2011-10-01

Full Text Available Abstract Background Hydrolysates of plant biomass used for the production of lignocellulosic biofuels typically contain sugar mixtures consisting mainly of D-glucose and D-xylose, and minor amounts of L-arabinose. The yeast Saccharomyces cerevisiae is the preferred microorganism for the fermentative production of ethanol but is not able to ferment pentose sugars. Although D-xylose and L-arabinose fermenting S. cerevisiae strains have been constructed recently, pentose uptake is still a limiting step in mixed sugar fermentations. Results Here we described the cloning and characterization of two sugar transporters, AraT from the yeast Scheffersomyces stipitis and Stp2 from the plant Arabidopsis thaliana, which mediate the uptake of L-arabinose but not of D-glucose into S. cerevisiae cells. A yeast strain lacking all of its endogenous hexose transporter genes and expressing a bacterial L-arabinose utilization pathway could no longer take up and grow with L-arabinose as the only carbon source. Expression of the heterologous transporters supported uptake and utilization of L-arabinose especially at low L-arabinose concentrations but did not, or only very weakly, support D-glucose uptake and utilization. In contrast, the S. cerevisiae D-galactose transporter, Gal2, mediated uptake of both L-arabinose and D-glucose, especially at high concentrations. Conclusions Using a newly developed screening system we have identified two heterologous sugar transporters from a yeast and a plant which can support uptake and utilization of L-arabinose in L-arabinose fermenting S. cerevisiae cells, especially at low L-arabinose concentrations.

Inhibitor tolerance of a recombinant flocculating industrial Saccharomyces cerevisiae strain during glucose and xylose co-fermentation

Directory of Open Access Journals (Sweden)

Yun-Cheng Li

Full Text Available ABSTRACT Lignocellulose-derived inhibitors have negative effects on the ethanol fermentation capacity of Saccharomyces cerevisiae. In this study, the effects of eight typical inhibitors, including weak acids, furans, and phenols, on glucose and xylose co-fermentation of the recombinant xylose-fermenting flocculating industrial S. cerevisiae strain NAPX37 were evaluated by batch fermentation. Inhibition on glucose fermentation, not that on xylose fermentation, correlated with delayed cell growth. The weak acids and the phenols showed additive effects. The effect of inhibitors on glucose fermentation was as follows (from strongest to weakest: vanillin > phenol > syringaldehyde > 5-HMF > furfural > levulinic acid > acetic acid > formic acid. The effect of inhibitors on xylose fermentation was as follows (from strongest to weakest: phenol > vanillin > syringaldehyde > furfural > 5-HMF > formic acid > levulinic acid > acetic acid. The NAPX37 strain showed substantial tolerance to typical inhibitors and showed good fermentation characteristics, when a medium with inhibitor cocktail or rape straw hydrolysate was used. This research provides important clues for inhibitors tolerance of recombinant industrial xylose-fermenting S. cerevisiae.

Identification and regulation of genes involved in anaerobic growth of Saccharomyces cerevisiae

NARCIS (Netherlands)

Snoek, Isidora Sophia Ishtar

2007-01-01

Saccharomyces cerevisiae is one of the few yeast species that can grow equally well without molecular oxygen (anaerobic) as with this compound present (aerobic). This property has made it one of the most abundantly used yeasts in industry, since anaerobic incubation plays a major part in alcohol and

Increased xylose affinity of Hxt2 through gene shuffling of hexose transporters in Saccharomyces cerevisiae

NARCIS (Netherlands)

Nijland, Jeroen G; Shin, Hyun Yong; de Waal, Paul P; Klaassen, Paul; Driessen, Arnold J M

AIMS: Optimizing D-xylose transport in Saccharomyces cerevisiae is essential for efficient bioethanol production from cellulosic materials. We have used a gene shuffling approach of hexose (Hxt) transporters in order to increase the affinity for D-xylose. METHODS AND RESULTS: Various libraries were

Xylose reductase from the thermophilic fungus Talaromyces emersonii

Indian Academy of Sciences (India)

Prakash

Xylose reductase is involved in the first step of the fungal pentose catabolic pathway. The gene .... proteins with reversed coenzyme preference from NADPH to NADH ..... 399–404. Hasper A A, Visser J and de Graaff L H 2000 The Aspergillus.

Small intestinal malabsorption in chronic alcoholism: a retrospective study of alcoholic patients by the ¹⁴C-D-xylose breath test.

Science.gov (United States)

Hope, Håvar; Skar, Viggo; Sandstad, Olav; Husebye, Einar; Medhus, Asle W

2012-04-01

The ¹⁴C-D-xylose breath test was used at Ullevål University Hospital in the period from 1986 TO 1995 for malabsorption testing. The objective of this retrospective study was to reveal whether patients with chronic alcoholism may have intestinal malabsorption. The consecutive ¹⁴C-D-xylose breath test database was reviewed and patients with the diagnosis of chronic alcoholism were identified. ¹⁴C-D-xylose breath test results of the alcoholic patients were compared with the results of untreated celiac patients and patient and healthy controls. In the ¹⁴C-D-xylose breath test, ¹⁴C-D-xylose was dissolved in water and given orally after overnight fast. Breath samples were taken at 30-min intervals for 210 min, and ¹⁴CO₂ : ¹²CO₂ ratios were calculated for each time point, presenting a time curve for ¹⁴C-D-xylose absorption. Urine was collected after 210 min and the fraction of the total d-xylose passed was calculated (U%). ¹⁴CO₂ in breath and ¹⁴C-D-xylose in urine were analyzed using liquid scintillation. Both breath and urine analysis revealed a pattern of malabsorption in alcoholics comparable with untreated celiac patients, with significantly reduced absorption of d-xylose compared with patient and healthy controls. Alcoholic patients have a significantly reduced ¹⁴C-D-xylose absorption, comparable with untreated celiac patients. This indicates a reduced intestinal function in chronic alcoholism.

Expression of protein engineered NADP{sup +}-dependent xylitol dehydrogenase increases ethanol production from xylose in recombinant Saccharomyces cerevisiae

Energy Technology Data Exchange (ETDEWEB)

Matsushika, Akinori; Inoue, Hiroyuki; Murakami, Katsuji; Takimura, Osamu; Sawayama, Shigeki [National Institute of Advanced Industrial Science and Technology, Hiroshima (Japan). Biomass Technology Research Center; Watanabe, Seiya; Kodaki, Tsutomu; Makino, Keisuke [Kyoto Univ. (Japan). Inst. of Advanced Energy

2008-11-15

A recombinant Saccharomyces cerevisiae strain transformed with xylose reductase (XR) and xylitol dehydrogenase (XDH) genes from Pichia stipitis has the ability to convert xylose to ethanol together with the unfavorable excretion of xylitol, which may be due to cofactor imbalance between NADPH-preferring XR and NAD{sup +}-dependent XDH. To reduce xylitol formation, we have already generated several XDH mutants with a reversal of coenzyme specificity toward NADP{sup +}. In this study, we constructed a set of recombinant S. cerevisiae strains with xylose-fermenting ability, including protein-engineered NADP{sup +}-dependent XDH-expressing strains. The most positive effect on xylose-to-ethanol fermentation was found by using a strain named MA-N5, constructed by chromosomal integration of the gene for NADP{sup +}-dependent XDH along with XR and endogenous xylulokinase genes. The MA-N5 strain had an increase in ethanol production and decrease in xylitol excretion compared with the reference strain expressing wild-type XDH when fermenting not only xylose but also mixed sugars containing glucose and xylose. Furthermore, the MA-N5 strain produced ethanol with a high yield of 0.49 g of ethanol/g of total consumed sugars in the nonsulfuric acid hydrolysate of wood chips. The results demonstrate that glucose and xylose present in the lignocellulosic hydrolysate can be efficiently fermented by this redox-engineered strain. (orig.)

Dehydration of xylose to furfural over MCM-41-supported niobium-oxide catalysts.

Science.gov (United States)

García-Sancho, Cristina; Sádaba, Irantzu; Moreno-Tost, Ramón; Mérida-Robles, Josefa; Santamaría-González, José; López-Granados, Manuel; Maireles-Torres, Pedro

2013-04-01

A series of silica-based MCM-41-supported niobium-oxide catalysts are prepared, characterized by using XRD, N2 adsorption-desorption, X-ray photoelectron spectroscopy, Raman spectroscopy, and pyridine adsorption coupled to FTIR spectroscopy, and tested for the dehydration of D-xylose to furfural. Under the operating conditions used all materials are active in the dehydration of xylose to furfural (excluding the MCM-41 silica support). The xylose conversion increases with increasing Nb2 O5 content. At a loading of 16 wt % Nb2 O5 , 74.5 % conversion and a furfural yield of 36.5 % is achieved at 170 °C, after 180 min reaction time. Moreover, xylose conversion and furfural yield increase with the reaction time and temperature, attaining 82.8 and 46.2 %, respectively, at 190 °C and after 100 min reaction time. Notably, the presence of NaCl in the reaction medium further increases the furfural yield (59.9 % at 170 °C after 180 min reaction time). Moreover, catalyst reutilization is demonstrated by performing at least three runs with no loss of catalytic activity and without the requirement for an intermediate regeneration step. No significant niobium leaching is observed, and a relationship between the structure of the catalyst and the activity is proposed. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Effects of alternative dietary substrates on competition between human colonic bacteria in an anaerobic fermentor system.

Science.gov (United States)

Duncan, Sylvia H; Scott, Karen P; Ramsay, Alan G; Harmsen, Hermie J M; Welling, Gjalt W; Stewart, Colin S; Flint, Harry J

2003-02-01

Duplicate anaerobic fermentor systems were used to examine changes in a community of human fecal bacteria supplied with different carbohydrate energy sources. A panel of group-specific fluorescent in situ hybridization probes targeting 16S rRNA sequences revealed that the fermentors supported growth of a greater proportion of Bacteroides and a lower proportion of gram-positive anaerobes related to Faecalibacterium prausnitzii, Ruminococcus flavefaciens-Ruminococcus bromii, Eubacterium rectale-Clostridium coccoides, and Eubacterium cylindroides than the proportions in the starting fecal inoculum. Nevertheless, certain substrates, such as dahlia inulin, caused a pronounced increase in the number of bacteria related to R. flavefaciens-R. bromii and E. cylindroides. The ability of three strictly anaerobic, gram-positive bacteria to compete with the complete human fecal flora was tested in the same experiment by using selective plating to enumerate the introduced strains. The Roseburia-related strain A2-183(F) was able to grow on all substrates despite the fact that it was unable to utilize complex carbohydrates in pure culture, and it was assumed that this organism survived by cross-feeding. In contrast, Roseburia intestinalis L1-82(R) and Eubacterium sp. strain A2-194(R) survived less well despite the fact that they were able to utilize polysaccharides in pure culture, except that A2-194(R) was stimulated 100-fold by inulin. These results suggest that many low-G+C-content gram-positive obligate anaerobes may be selected against during in vitro incubation, although several groups were stimulated by inulin. Thus, considerable caution is necessary when workers attempt to predict the in vivo effects of probiotics and prebiotics from their effects in vitro.

Acid-catalysed xylose dehydration into furfural in the presence of kraft lignin.

Science.gov (United States)

Lamminpää, Kaisa; Ahola, Juha; Tanskanen, Juha

2015-02-01

In this study, the effects of kraft lignin (Indulin AT) on acid-catalysed xylose dehydration into furfural were studied in formic and sulphuric acids. The study was done using D-optimal design. Three variables in both acids were included in the design: time (20-80 min), temperature (160-180°C) and initial lignin concentration (0-20 g/l). The dependent variables were xylose conversion, furfural yield, furfural selectivity and pH change. The results showed that the xylose conversion and furfural yield decreased in sulphuric acid, while in formic acid the changes were minor. Additionally, it was showed that lignin has an acid-neutralising capacity, and the added lignin increased the pH of reactant solutions in both acids. The pH rise was considerably lower in formic acid than in sulphuric acid. However, the higher pH did not explain all the changes in conversion and yield, and thus lignin evidently inhibits the formation of furfural. Copyright © 2014 Elsevier Ltd. All rights reserved.

KINETICS OF GROWTH AND ETHANOL PRODUCTION ON DIFFERENT CARBON SUBSTRATES USING GENETICALLY ENGINEERED XYLOSE-FERMENTING YEAST

Science.gov (United States)

Saccharomyces cerevisiae 424A (LNH-ST) strain was used for fermentation of glucose and xylose. Growth kinetics and ethanol productivity were calculated for batch fermentation on media containing different combinations of glucose and xylose to give a final sugar concentra...

Anaerobic membrane bio-reactors for severe industrial effluents and urban spill waters : The AMBROSIUS project

NARCIS (Netherlands)

Van Lier, J.B.; Ozgun, H.; Ersahin, M.E.; Dereli, R.K.

2013-01-01

With growing application experiences from aerobic membrane bioreactors, combination of membrane and anaerobic processes become more and more attractive and feasible. In anaerobic membrane bioreactors (AnMBRs), biomass and particulate organic matter are physically retained inside the reactor,

Vaspar broth-disk procedure for antibiotic susceptibility testing of anaerobic bacteria.

OpenAIRE

West, S E; Wilkins, T D

1980-01-01

A modification of the Wilkins-Thiel broth-disk procedure for antibiotic susceptibility testing of anaerobic bacteria is described. This method utilizes an aerobically prepared medium overlaid with molten vaspar. Specialized anaerobic techniques or prereduced media are not required.

The structure of apo and holo forms of xylose reductase, a dimeric aldo-keto reductase from Candida tenuis.

Science.gov (United States)

Kavanagh, Kathryn L; Klimacek, Mario; Nidetzky, Bernd; Wilson, David K

2002-07-16

Xylose reductase is a homodimeric oxidoreductase dependent on NADPH or NADH and belongs to the largely monomeric aldo-keto reductase superfamily of proteins. It catalyzes the first step in the assimilation of xylose, an aldose found to be a major constituent monosaccharide of renewable plant hemicellulosic material, into yeast metabolic pathways. It does this by reducing open chain xylose to xylitol, which is reoxidized to xylulose by xylitol dehydrogenase and metabolically integrated via the pentose phosphate pathway. No structure has yet been determined for a xylose reductase, a dimeric aldo-keto reductase or a family 2 aldo-keto reductase. The structures of the Candida tenuis xylose reductase apo- and holoenzyme, which crystallize in spacegroup C2 with different unit cells, have been determined to 2.2 A resolution and an R-factor of 17.9 and 20.8%, respectively. Residues responsible for mediating the novel dimeric interface include Asp-178, Arg-181, Lys-202, Phe-206, Trp-313, and Pro-319. Alignments with other superfamily members indicate that these interactions are conserved in other dimeric xylose reductases but not throughout the remainder of the oligomeric aldo-keto reductases, predicting alternate modes of oligomerization for other families. An arrangement of side chains in a catalytic triad shows that Tyr-52 has a conserved function as a general acid. The loop that folds over the NAD(P)H cosubstrate is disordered in the apo form but becomes ordered upon cosubstrate binding. A slow conformational isomerization of this loop probably accounts for the observed rate-limiting step involving release of cosubstrate. Xylose binding (K(m) = 87 mM) is mediated by interactions with a binding pocket that is more polar than a typical aldo-keto reductase. Modeling of xylose into the active site of the holoenzyme using ordered waters as a guide for sugar hydroxyls suggests a convincing mode of substrate binding.

Characteristics and performance of anaerobic wastewater treatment (a review)

International Nuclear Information System (INIS)

Zeb, B.S.; Mahmood, Q.; Pervez, A.

2013-01-01

Summary: Pakistan's current population of 180 million is expected to grow to about 221 million by the year 2025. In developing countries such as Pakistan water pollution is a major threat to the livelihood of people. Pakistan is also currently experiencing profound demographic, economic changes and energy crisis that have major implications for water management. The contamination of aquatic and terrestrial ecosystems with heavy metals is a major environmental problem. Each pollution problem calls for specific optimal and cost effective solution so if one technology proves less or ineffective other takes its place. Every day the vast amounts of the municipal, industrial and agricultural wastes are released in to the environment and create serious problems. Anaerobic digestion is very attractive and cost-effective option and technology for the highly loaded waste water treatment and energy conversion. The anaerobic process is in many ways ideal for waste treatment. It has several significant advantages over other available methods. In this process organic matter is utilized as source of electron donor to reduce carbon dioxide to produce methane gas. It involves three bacterial groups namely: hydrolytic, acetogenic and methanogenic bacteria that work optimally at pH and temperature ranges of 6.8 to 7.5 and 30-35 degree C, respectively. The residence time in a digester varies with the amount and type of feed material, the configuration of the digestion system, and whether it be one-stage or two-stage. It is ideal for all kinds of wastewaters. Currently anaerobic technology is being operated at full scale in many industrialized nations. (author)

Effects of furfural and acetic acid on growth and lipid production from glucose and xylose by Rhodotorula glutinis

Energy Technology Data Exchange (ETDEWEB)

Zhang, Guochang; French, William Todd; Hernandez, Rafael; Alley, Earl; Paraschivescu, Maria [Dave C. Swalm School of Chemical Engineering, Mississippi State University, P.O. Box 9595, Mississippi State, MS 39762 (United States)

2011-01-15

Microbial conversion of lignocellulosic sugars to triacylglycerols (a biodiesel or renewable diesel feedstock) was investigated using the oleaginous yeast Rhodotorula glutinis (ATCC 15125). In the shake flask experiments, R. glutinis was first grown in a nitrogen-rich medium utilizing an artificial acid hydrolysate of lignocellulosic biomass switchgrass as the sole carbon and energy source. Once the culture had reached the stationary phase, the cells were harvested and transferred to a fresh nitrogen-free media containing artificial acid hydrolysate sugars for lipid accumulation. Analysis of the data collected showed that the yeast were able to grow in the medium containing artificial acid hydrolysate sugars as the carbon and energy source. The net specific Growth rate(s) indicated that the presence of acetic acid and furfural in the artificial acid hydrolysate inhibited the growth of R. glutinis on glucose, but not the growth on xylose. The lipid accumulated in the cells, determined by gravimetrical method, increased from initial 4.3%-39.0% of dry cell mass weight. The major fatty acids of the accumulated lipids were palmitic acid, stearic acid, oleic acid, linoleic acid and {gamma}-linoleic acid. These results indicate that it is feasible to convert the sugars in acid hydrolysate of lignocellulosic biomass to triacylglycerols using R. glutinis. (author)

Evidence supporting dissimilatory and assimilatory lignin degradation in Enterobacter lignolyticus SCF1

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Kristen M DeAngelis

2013-09-01

Full Text Available The anaerobic isolate Enterobacter lignolyticus SCF1 was initially cultivated based on anaerobic growth on lignin as sole carbon source. The source of the isolated bacteria was from tropical forest soils that decompose litter rapidly with low and fluctuating redox potentials, making it likely that bacteria using oxygen-independent enzymes play an important role in decomposition. We have used transcriptomics and proteomics to examine the increased growth of the anaerobic isolate Enterobacter lignolyticus SCF1 when grown on media amended with lignin compared to unamended growth. Proteomics revealed accelerated xylose uptake and metabolism under lignin-amended growth, and lignin degradation via the 4-hydroxyphenylacetate degradation pathway, catalase/peroxidase enzymes, and the glutathione biosynthesis and glutathione S-transferase proteins. We also observed increased production of NADH-quinone oxidoreductase, other electron transport chain proteins, and ATP synthase and ATP-binding cassette (ABC transporters. We detected significant lignin degradation over time by absorbance, and also used metabolomics to demonstrate increased xylose utilization in lignin-amended compared to unamended growth. Our data shows the advantages of a multi-omics approach, where incomplete pathways identified by genomics were completed, and new observations made on coping with poor carbon availability. The fast growth, high efficiency and specificity of enzymes employed in bacterial anaerobic litter deconstruction makes these soils useful templates for improving biofuel production.

Diversity and physiological characterization of D-xylose-fermenting yeasts isolated from the Brazilian Amazonian Forest.

Science.gov (United States)

Cadete, Raquel M; Melo, Monaliza A; Dussán, Kelly J; Rodrigues, Rita C L B; Silva, Silvio S; Zilli, Jerri E; Vital, Marcos J S; Gomes, Fátima C O; Lachance, Marc-André; Rosa, Carlos A

2012-01-01

This study is the first to investigate the Brazilian Amazonian Forest to identify new D-xylose-fermenting yeasts that might potentially be used in the production of ethanol from sugarcane bagasse hemicellulosic hydrolysates. A total of 224 yeast strains were isolated from rotting wood samples collected in two Amazonian forest reserve sites. These samples were cultured in yeast nitrogen base (YNB)-D-xylose or YNB-xylan media. Candida tropicalis, Asterotremella humicola, Candida boidinii and Debaryomyces hansenii were the most frequently isolated yeasts. Among D-xylose-fermenting yeasts, six strains of Spathaspora passalidarum, two of Scheffersomyces stipitis, and representatives of five new species were identified. The new species included Candida amazonensis of the Scheffersomyces clade and Spathaspora sp. 1, Spathaspora sp. 2, Spathaspora sp. 3, and Candida sp. 1 of the Spathaspora clade. In fermentation assays using D-xylose (50 g/L) culture medium, S. passalidarum strains showed the highest ethanol yields (0.31 g/g to 0.37 g/g) and productivities (0.62 g/L · h to 0.75 g/L · h). Candida amazonensis exhibited a virtually complete D-xylose consumption and the highest xylitol yields (0.55 g/g to 0.59 g/g), with concentrations up to 25.2 g/L. The new Spathaspora species produced ethanol and/or xylitol in different concentrations as the main fermentation products. In sugarcane bagasse hemicellulosic fermentation assays, S. stipitis UFMG-XMD-15.2 generated the highest ethanol yield (0.34 g/g) and productivity (0.2 g/L · h), while the new species Spathaspora sp. 1 UFMG-XMD-16.2 and Spathaspora sp. 2 UFMG-XMD-23.2 were very good xylitol producers. This study demonstrates the promise of using new D-xylose-fermenting yeast strains from the Brazilian Amazonian Forest for ethanol or xylitol production from sugarcane bagasse hemicellulosic hydrolysates.

Meta-analysis and functional validation of nutritional requirements of solventogenic Clostridia growing under butanol stress conditions and coutilization of D-glucose and D-xylose.

Science.gov (United States)

Heluane, Humberto; Evans, Matthew R; Dagher, Sue F; Bruno-Bárcena, José M

2011-07-01

Recent advances in systems biology, omics, and computational studies allow us to carry out data mining for improving biofuel production bioprocesses. Of particular interest are bioprocesses that center on microbial capabilities to biotransform both the hexose and pentose fractions present in crop residues. This called for a systematic exploration of the components of the media to obtain higher-density cultures and more-productive fermentation operations than are currently found. By using a meta-analysis approach of the transcriptional responses to butanol stress, we identified the nutritional requirements of solvent-tolerant strain Clostridium beijerinckii SA-1 (ATCC 35702). The nutritional requirements identified were later validated using the chemostat pulse-and-shift technique. C. beijerinckii SA-1 was cultivated in a two-stage single-feed-stream continuous production system to test the proposed validated medium formulation, and the coutilization of D-glucose and D-xylose was evaluated by taking advantage of the well-known ability of solventogenic clostridia to utilize a large variety of carbon sources such as mono-, oligo-, and polysaccharides containing pentose and hexose sugars. Our results indicated that C. beijerinckii SA-1 was able to coferment hexose/pentose sugar mixtures in the absence of a glucose repression effect. In addition, our analysis suggests that the solvent and acid resistance mechanisms found in this strain are differentially regulated compared to strain NRRL B-527 and are outlined as the basis of the analysis toward optimizing butanol production.

The genome sequences of Cellulomonas fimi and "Cellvibrio gilvus" reveal the cellulolytic strategies of two facultative anaerobes, transfer of "Cellvibrio gilvus" to the genus Cellulomonas, and proposal of Cellulomonas gilvus sp. nov.

Directory of Open Access Journals (Sweden)

Melissa R Christopherson

Full Text Available Actinobacteria in the genus Cellulomonas are the only known and reported cellulolytic facultative anaerobes. To better understand the cellulolytic strategy employed by these bacteria, we sequenced the genome of the Cellulomonas fimi ATCC 484(T. For comparative purposes, we also sequenced the genome of the aerobic cellulolytic "Cellvibrio gilvus" ATCC 13127(T. An initial analysis of these genomes using phylogenetic and whole-genome comparison revealed that "Cellvibrio gilvus" belongs to the genus Cellulomonas. We thus propose to assign "Cellvibrio gilvus" to the genus Cellulomonas. A comparative genomics analysis between these two Cellulomonas genome sequences and the recently completed genome for Cellulomonas flavigena ATCC 482(T showed that these cellulomonads do not encode cellulosomes but appear to degrade cellulose by secreting multi-domain glycoside hydrolases. Despite the minimal number of carbohydrate-active enzymes encoded by these genomes, as compared to other known cellulolytic organisms, these bacteria were found to be proficient at degrading and utilizing a diverse set of carbohydrates, including crystalline cellulose. Moreover, they also encode for proteins required for the fermentation of hexose and xylose sugars into products such as ethanol. Finally, we found relatively few significant differences between the predicted carbohydrate-active enzymes encoded by these Cellulomonas genomes, in contrast to previous studies reporting differences in physiological approaches for carbohydrate degradation. Our sequencing and analysis of these genomes sheds light onto the mechanism through which these facultative anaerobes degrade cellulose, suggesting that the sequenced cellulomonads use secreted, multidomain enzymes to degrade cellulose in a way that is distinct from known anaerobic cellulolytic strategies.

Simultaneous Decolorization and Biohydrogen Production from Xylose by Klebsiella oxytoca GS-4-08 in the Presence of Azo Dyes with Sulfonate and Carboxyl Groups

Science.gov (United States)

Cao, Ming-yue; Wang, Peng-tao; Wang, Shi; Yue, Ying-rong; Yuan, Wen-duo; Qiao, Wei-chuan; Wang, Fei

2017-01-01

ABSTRACT Biohydrogen production from the pulp and paper effluent containing rich lignocellulosic material could be achieved by the fermentation process. Xylose, an important hemicellulose hydrolysis product, is used less efficiently as a substrate for biohydrogen production. Moreover, azo dyes are usually added to fabricate anticounterfeiting paper, which further increases the complexity of wastewater. This study reports that xylose could serve as the sole carbon source for a pure culture of Klebsiella oxytoca GS-4-08 to achieve simultaneous decolorization and biohydrogen production. With 2 g liter−1 of xylose as the substrate, a maximum xylose utilization rate (URxyl) and a hydrogen molar yield (HMY) of 93.99% and 0.259 mol of H2 mol of xylose−1, respectively, were obtained. Biohydrogen kinetics and electron equivalent (e− equiv) balance calculations indicated that methyl red (MR) penetrates and intracellularly inhibits both the pentose phosphate pathway and pyruvate fermentation pathway, while methyl orange (MO) acted independently of the glycolysis and biohydrogen pathway. The data demonstrate that biohydrogen pathways in the presence of azo dyes with sulfonate and carboxyl groups were different, but the azo dyes could be completely reduced during the biohydrogen production period in the presence of MO or MR. The feasibility of hydrogen production from industrial pulp and paper effluent by the strain if the xylose is sufficient was also proved and was not affected by toxic substances which usually exist in such wastewater, except for chlorophenol. This study offers a promising energy-recycling strategy for treating pulp and paper wastewaters, especially for those containing azo dyes. IMPORTANCE The pulp and paper industry is a major industry in many developing countries, and the global market of pulp and paper wastewater treatment is expected to increase by 60% between 2012 and 2020. Such wastewater contains large amounts of refractory contaminants, such

Optimization studies on acid hydrolysis of oil palm empty fruit bunch fiber for production of xylose.

Science.gov (United States)

Rahman, S H A; Choudhury, J P; Ahmad, A L; Kamaruddin, A H

2007-02-01

Oil palm empty fruit bunch fiber is a lignocellulosic waste from palm oil mills. It is a potential source of xylose which can be used as a raw material for production of xylitol, a high value product. The increasing interest on use of lignocellulosic waste for bioconversion to fuels and chemicals is justifiable as these materials are low cost, renewable and widespread sources of sugars. The objective of the present study was to determine the effect of H(2)SO(4) concentration, reaction temperature and reaction time for production of xylose. Batch reactions were carried out under various reaction temperature, reaction time and acid concentrations and Response Surface Methodology (RSM) was followed to optimize the hydrolysis process in order to obtain high xylose yield. The optimum reaction temperature, reaction time and acid concentration found were 119 degrees C, 60 min and 2%, respectively. Under these conditions xylose yield and selectivity were found to be 91.27% and 17.97 g/g, respectively.

Metabolic control analysis of xylose catabolism in Aspergillus

DEFF Research Database (Denmark)

Prathumpai, Wai; Gabelgaard, J.B.; Wanchanthuek, P.

2003-01-01

, and flux control was shown to be dependent on the metabolite levels. Due to thermodynamic constraints, flux control may reside at the first step in the pathway, i.e., at the xylose reductase, even when the intracellular xylitol concentration is high. On the basis of the kinetic analysis, the general dogma...

Site-specific management of miscanthus genotypes for combustion and anaerobic digestion

NARCIS (Netherlands)

Kiesel, Andreas; Nunn, Christopher; Iqbal, Yasir; Weijde, Van der Tim; Wagner, Moritz; Özgüven, Mensure; Tarakanov, Ivan; Kalinina, Olena; Trindade, Luisa M.; Clifton-Brown, John; Lewandowski, Iris

2017-01-01

In Europe, the perennial C4 grass miscanthus is currently mainly cultivated for energy generation via combustion. In recent years, anaerobic digestion has been identified as a promising alternative utilization pathway. Anaerobic digestion produces a higher-value intermediate (biogas), which can be

Effect of methanogenic substrates on anaerobic oxidation of methane and sulfate reduction by an anaerobic methanotrophic enrichment.

KAUST Repository

Meulepas, Roel J W; Jagersma, Christian G; Khadem, Ahmad F; Stams, Alfons J M; Lens, Piet N L

2010-01-01

Anaerobic oxidation of methane (AOM) coupled to sulfate reduction (SR) is assumed to be a syntrophic process, in which methanotrophic archaea produce an interspecies electron carrier (IEC), which is subsequently utilized by sulfate-reducing bacteria

Physiologically anaerobic microorganisms of the deep subsurface

International Nuclear Information System (INIS)

Stevens, S.E. Jr.; Chung, K.T.

1993-10-01

Anaerobic bacteria were isolated from deep subsurface sediment samples taken at study sites in Idaho (INEL) and Washington (HR) by culturing on dilute and concentrated medium. Morphologically distinct colonies were purified, and their responses to 21 selected physiological tests were determined. Although the number of isolates was small (18 INEL, 27 HR) some general patterns could be determined. Most strains could utilize all the carbon sources, however the glycerol and melizitose utilization was positive for 50% or less of the HR isolates. Catalase activity (27.78% at INEL, 74.07% at HR) and tryptophan metabolism (11.12% at INEL, 40.74% at HR) were significantly different between the two study sites. MPN and viable counts indicate that sediments near the water table yield the greatest numbers of anaerobes. Deeper sediments also appear to be more selective with the greatest number of viable counts on low-nutrient mediums. Likewise, only strictly obligate anaerobes were found in the deepest sediment samples. Selective media indicated the presence of methanogens, acetogens, and sulfate reducers at only the HR site

CHARACTERIZATION OF AN ANAEROBIC FUNGUS FROM LLAMA FECES

NARCIS (Netherlands)

MARVINSIKKEMA, FD; LAHPOR, GA; KRAAK, MN; GOTTSCHAL, JC; PRINS, RA

1992-01-01

An anaerobic fungus was isolated from Hama faeces. Based on its morphological characteristics, polyflagellated zoospores, extensive rhizoid system and the formation of monocentric colonies, the fungus is assigned to the genus Neocallimastix. Neocallimastix sp. L2 is able to grow on several poly-,

Optimization of hydrothermal pretreatment for co-utilization C-5 and C-6 sugars of cassava alcohol residue

International Nuclear Information System (INIS)

Lu, Huisheng; Lv, Chunliu; Zhang, Minhua; Liu, Shuangyan; Liu, Jiatao; Lian, Feng

2017-01-01

Highlights: • Cassava alcohol residue was first pretreated by hydrothermal reaction. • Hydrothermal pretreatment was optimized by RSM for co-utilization of C-5 and C-6 sugars. • The maximum xylose yield and the highest enzymatic digestibility were not obtained at the same conditions. • Optimum pretreatment conditions were at 193 °C, with 11.4% solids and for 51 min. • The optimal theoretical ethanol production was 69.5 mg/g raw materials by co-utilization of C-5 and C-6 sugars. - Abstract: Hydrothermal reaction was first applied to pretreat cassava alcohol residue for realizing the co-utilization of xylose and glucose to assume fermentation ethanol. This work focused on the influence of hydrothermal pretreatment conditions on ethanol production. Hydrothermal reaction was used to explore the maximum xylose and glucose yields, in respect to reaction temperature (120–240 °C), solid-liquid ratio (0.023–0.150) and reaction time (15–120 min). The results showed that the suitable conditions were at 180–200 °C, for 45–60 min and with 10–12.5% solids. In this range, the conjunct of xylose and glucose would reach the maximum, which can make full use of hemicellulose and cellulose in cassava alcohol residue. According to the results, respond surface methodology (RSM) based on Box-Behnken design was used to further optimize the three independent variables for the highest ethanol by co-utilization of xylose and glucose. RSM revealed that the effect of temperature on ethanol production was much more significant than the effect of reaction time and solid-liquid ratio, and the highest ethanol production was 70.6 mg/g that was close to the experiment value of 69.5 mg/g at 193 °C for 51 min and with 11.5% solids. Furthermore, the crystallinity and morphology of the untreated and pretreated cassava alcohol residue were investigated to assess the effect of hydrothermal pretreatment by scanning electron microscope (SEM) and X-ray diffraction (XRD

Silencing ß1,2-xylosyltransferase in transgenic tomato fruits reveals xylose as constitutive component of IgE binding epitopes

Directory of Open Access Journals (Sweden)

Kathrin Elisabeth Paulus

2011-08-01

Full Text Available Complex plant N-glycans containing β1,2-xylose and core α1,3-fucose are regarded as the major class of the so-called ‘carbohydrate cross-reactive determinants’ reactive with IgE antibodies in sera of many allergic patients, but their clinical relevance is still under debate. Plant glycosyltransferases, β1,2-xylosyltransferase (XylT and core α1,3-fucosyltransferase (FucT are responsible for the transfer of β1,2-linked xylose and core α1,3-linked fucose residues to N-glycans of glycoproteins, respectively. To test the clinical relevance of ß 1,2-xylose containing epitopes, expression of the tomato β1,2-xylosyltransferase was down-regulated by RNA interference (RNAi in transgenic plants. Fruits harvested from these transgenic plants were analysed for accumulation of XylT mRNA, abundance of ß1,2-xylose epitopes and their allergenic potential. Based on qPCR analysis XylT mRNA levels were reduced up to 10-fold in independent transgenic lines as compared to untransformed control, whereas no xylosylated N-glycans could be revealed by MS analysis. Immunoblotting using anti-xylose-specific IgG antibodies revealed a strong reduction of ß1,2-xylose containing epitopes. Incubating protein extracts from untransformed controls and XylT_RNAi plants with sera from tomato allergic patients showed a patient-specific reduction in IgE binding, indicating a reduced allergenic potential of XylT_RNAi tomato fruits, in vitro. To elucidate the clinical relevance of ß1,2-xylose containing complex N-glycans skin prick tests were performed demonstrating a reduced responsiveness of tomato allergic patients, in vivo. This study provides strong evidence for the clinical relevance of ß1,2-xylose containing epitopes in vivo.

Identification of a conserved protein involved in anaerobic unsaturated fatty acid synthesis in Neiserria gonorrhoeae: implications for facultative and obligate anaerobes that lack FabA

Science.gov (United States)

Isabella, Vincent M.; Clark, Virginia L.

2011-01-01

SUMMARY Transcriptome analysis of the facultative anaerobe, Neisseria gonorrhoeae, revealed that many genes of unknown function were induced under anaerobic conditions. Mutation of one such gene, NGO1024, encoding a protein belonging to the 2-nitropropane dioxygenase-like superfamiliy of proteins, was found to result in an inability of gonococci to grow anaerobically. Anaerobic growth of an NG1024 mutant was restored upon supplementation with unsaturated fatty acids (UFA), but not with the saturated fatty acid palmitate. Gonococcal fatty acid profiles confirmed that NGO1024 was involved in UFA synthesis anaerobically, but not aerobically, demonstrating that gonococci contain two distinct pathways for the production of UFAs, with a yet unidentified aerobic mechanism, and an anaerobic mechanism involving NGO1024. Expression of genes involved in classical anaerobic UFA synthesis, fabA, fabM, and fabB, was toxic in gonococci and unable to complement a NGO1024 mutation, suggesting that the chemistry involved in gonococcal anaerobic UFA synthesis is distinct from that of the classical pathway. NGO1024 homologs, which we suggest naming UfaA, form a distinct lineage within the 2-nitropropane dioxygenase-like superfamily, and are found in many facultative and obligate anaerobes that produce UFAs but lack fabA, suggesting that UfaA is part of a widespread pathway involved in UFA synthesis. PMID:21895795

Xylitol production from xylose mother liquor: a novel strategy that combines the use of recombinant Bacillus subtilis and Candida maltosa

OpenAIRE

Jiang Mingguo; Lv Jiyang; Wang Ben; Cheng Hairong; Lin Shuangjun; Deng Zixin

2011-01-01

Abstract Background Xylose mother liquor has high concentrations of xylose (35%-40%) as well as other sugars such as L-arabinose (10%-15%), galactose (8%-10%), glucose (8%-10%), and other minor sugars. Due to the complexity of this mother liquor, further isolation of xylose by simple method is not possible. In China, more than 50,000 metric tons of xylose mother liquor was produced in 2009, and the management of sugars like xylose that present in the low-cost liquor is a problem. Results We d...

Transport of D-xylose in Lactobacillus pentosus, Lactobacillus casei, and Lactobacillus plantarum: Evidence for a mechanism of facilitated diffusion via the phosphoenolpyruvate:mannose phosphotransferase system

NARCIS (Netherlands)

Chaillou, S.; Pouwels, P.H.; Postma, P.W.

1999-01-01

We have identified and characterized the D-xylose transport system of Lactobacillus pentosus. Uptake of D-xylose was not driven by the proton motive force generated by malolactic fermentation and required D-xylose metabolism. The kinetics of D-xylose transport were indicative of a low- affinity

Production of hemicellulose-degrading enzymes by Bacillus macerans in anaerobic culture

Energy Technology Data Exchange (ETDEWEB)

Williams, A.G.; Withers, S.E.

1985-09-01

The cell-associated and exocellular hemicellulolytic polysaccharide depolymerase and glycoside hydrolase activity of Bacillus macerans NCDO 1764 was monitored over a range of anaerobic growth conditions in batch and continuous culture. The enzymes were detectable throughout the complete growth cycle in batch culture reaching and maintaining maximum levels in the stationary phase. In continuous culture enzyme activity was largely independent of growth rate (D=0.025-0.1 h/sup -1/) although the activity was reduced at higher dilution rates (0.125-0.15 h/sup -1/). Although activity was detectable over a wide pH range (pH 5.5-7.5) it was pH dependent, and maximum activities of both the cell-associated and exocellular enzymes were measured in cultures maintained at pH 6.5-7.0 +- 0.1. The principal metabolites formed anaerobically from xylose by B. macerans in batch and continuous culture were acetic acid, formic acid and ethanol which represented 95-99% of the products formed. Smaller amounts of acetone, D,L-lactic acid and succinic acid were formed together with traces of butyric acid (<5 nmol/ml) and isovaleric acid (<25 nmol/ml). The proportions of the metabolites produced varied with growth conditions and were influenced by the pH of the culture and the rate and stage of growth of the microorganism.

Xylitol production from waste xylose mother liquor containing miscellaneous sugars and inhibitors: one-pot biotransformation by Candida tropicalis and recombinant Bacillus subtilis.

Science.gov (United States)

Wang, Hengwei; Li, Lijuan; Zhang, Lebin; An, Jin; Cheng, Hairong; Deng, Zixin

2016-05-16

The process of industrial xylitol production is a massive source of organic pollutants, such as waste xylose mother liquor (WXML), a viscous reddish-brown liquid. Currently, WXML is difficult to reuse due to its miscellaneous low-cost sugars, high content of inhibitors and complex composition. WXML, as an organic pollutant of hemicellulosic hydrolysates, accumulates and has become an issue of industrial concern in China. Previous studies have focused only on the catalysis of xylose in the hydrolysates into xylitol using one strain, without considering the removal of other miscellaneous sugars, thus creating an obstacle to subsequent large-scale purification. In the present study, we aimed to develop a simple one-pot biotransformation to produce high-purity xylitol from WXML to improve its economic value. In the present study, we developed a procedure to produce xylitol from WXML, which combines detoxification, biotransformation and removal of by-product sugars (purification) in one bioreactor using two complementary strains, Candida tropicalis X828 and Bacillus subtilis Bs12. At the first stage of micro-aerobic biotransformation, the yeast cells were allowed to grow and metabolized glucose and the inhibitors furfural and hydroxymethyl furfural (HMF), and converted xylose into xylitol. At the second stage of aerobic biotransformation, B. subtilis Bs12 was activated and depleted the by-product sugars. The one-pot process was successfully scaled up from shake flasks to 5, 150 L and 30 m(3) bioreactors. Approximately 95 g/L of pure xylitol could be obtained from the medium containing 400 g/L of WXML at a yield of 0.75 g/g xylose consumed, and the by-product sugars glucose, L-arabinose and galactose were depleted simultaneously. Our results demonstrate that the one-pot procedure is a viable option for the industrial application of WXML to produce value-added chemicals. The integration of complementary strains in the biotransformation of hemicellulosic hydrolysates is

Increasing ethanol productivity during xylose fermentation by cell recycling of recombinant Saccharomyces cerevisiae

DEFF Research Database (Denmark)

Roca, Christophe Francois Aime; Olsson, Lisbeth

2003-01-01

The influence of cell recycling of xylose-fermenting Saccharomyces cerevisiae TMB3001 was investigated during continuous cultivation on a xylose-glucose mixture. By using cell recycling at the dilution rate (D) of 0.05 h(-1), the cell-mass concentration could be increased from 2.2 g l(-1) to 22 g l...... ethanol productivity was in the range of 0.23-0.26 g g(-1) h(-1) with or without cell recycling, showing that an increased cell-mass concentration did not influence the efficiency of the yeast....

Screening and characterizing of xylanolytic and xylose-fermenting yeasts isolated from the wood-feeding termite, Reticulitermes chinensis.

Directory of Open Access Journals (Sweden)

Sameh Samir Ali

Full Text Available The effective fermentation of xylose remains an intractable challenge in bioethanol industry. The relevant xylanase enzyme is also in a high demand from industry for several biotechnological applications that inevitably in recent times led to many efforts for screening some novel microorganisms for better xylanase production and fermentation performance. Recently, it seems that wood-feeding termites can truly be considered as highly efficient natural bioreactors. The highly specialized gut systems of such insects are not yet fully realized, particularly, in xylose fermentation and xylanase production to advance industrial bioethanol technology as well as industrial applications of xylanases. A total of 92 strains from 18 yeast species were successfully isolated and identified from the gut of wood-feeding termite, Reticulitermes chinensis. Of these yeasts and strains, seven were identified for new species: Candida gotoi, Candida pseudorhagii, Hamamotoa lignophila, Meyerozyma guilliermondii, Sugiyamaella sp.1, Sugiyamaella sp. 2, and Sugiyamaella sp.3. Based on the phylogenetic and phenotypic characterization, the type strain of C. pseudorhagii sp. nov., which was originally designated strain SSA-1542T, was the most frequently occurred yeast from termite gut samples, showed the highly xylanolytic activity as well as D-xylose fermentation. The highest xylanase activity was recorded as 1.73 and 0.98 U/mL with xylan or D-xylose substrate, respectively, from SSA-1542T. Among xylanase-producing yeasts, four novel species were identified as D-xylose-fermenting yeasts, where the yeast, C. pseudorhagii SSA-1542T, showed the highest ethanol yield (0.31 g/g, ethanol productivity (0.31 g/L·h, and its fermentation efficiency (60.7% in 48 h. Clearly, the symbiotic yeasts isolated from termite guts have demonstrated a competitive capability to produce xylanase and ferment xylose, suggesting that the wood-feeding termite gut is a promising reservoir for novel

D-Xylose from waste liquors of a viscose process

Energy Technology Data Exchange (ETDEWEB)

Hashimoto, T; Mimura, M

1977-12-14

D-Xylose was prepared in good yields by neutralizing alkali waste liquors containing hemicellulose (I) with inorganic acids, dialyzing to remove salts hydrolyzing with acids, fermenting to decompose hexose, decolorizing, concentrating to < 15% sugars, treating with alcohols to precipitate oligosugars, removing the precipitate, and crystalizing. Thus, 1 kg waste liquor containing 27 g I was neutralized with 5% HCl, dialyzed at 15/sup 0/ for 48 h with parchment paper, concentrated at 40/sup 0/ to give a 500 g solution containing 7% H/sub 2/SO/sub 4/, boiled for 3 h, neutralized with BaCO/sub 3/, mixed with 10 g yeast at pH 5.4 to 5.8 (filtrate) fermented at 35/sup 0/ for 12 h, filtered, decolorized, concentrated at 40/sup 0/ to > 80 g mixed with EtOH to give a precipitate, filtered, concentrated to 17 g syrup, and mixed with AcOH to obtain 7.2 g D-Xylose.

Utilization of steam- and explosion-decompressed aspen wood by some anaerobes. [Acetivibrio cellulolyticus, Clostridium saccharolyticum, Zymomonas anaerobia

Energy Technology Data Exchange (ETDEWEB)

Khan, A W; Asther, M; Giuliano, C

1984-01-01

Tests made to study the suitability of using steam- and explosion-decompressed aspen wood as substrate in anaerobic fermentations indicated that after washing with dilute NaOH it becomes over 80% accessible to both mesophilic and thermophilic cellulolytic anaerobes and cellulases, compared with delignified, ball-milled pulp. After washing, this material was also found to be suitable for the single-step conversion of cellulose to ethanol using cocultures consisting of cellylolytic and ethanol-producing saccharolytic anaerobes; and without and after washing by the use of cellulolytic enzymes and ethanologenic anaerobes. 16 references, 3 tables.

Kinetic model for an up-flow anaerobic packed bed bioreactor: Dairy ...

African Journals Online (AJOL)

Kinetic studies of anaerobic digestion process of cheese whey were conducted in a pilot-scale up-flow anaerobic packed bed bioreactor (UAPB). An influent COD concentration of 59419 mg/l was utilized at steady state condition. Logistic and Monod kinetic models were employed to describe microbial activities of cheese ...

Wood-plastic composites utilizing wood flours derived from fast- growing trees common to the midwest

Science.gov (United States)

There are several non- or under-utilized hardwood trees common to the Midwestern states. Wood flour (WF) derived from fast-growing Midwest trees (Osage orange, Black Locust and Red Mulberry) were evaluated as a source of bio-based fiber reinforcements. Wood plastic composites (WPC) of high density p...

Single-cell Protein and Xylitol Production by a Novel Yeast Strain Candida intermedia FL023 from Lignocellulosic Hydrolysates and Xylose.

Science.gov (United States)

Wu, Jiaqiang; Hu, Jinlong; Zhao, Shumiao; He, Mingxiong; Hu, Guoquan; Ge, Xiangyang; Peng, Nan

2018-05-01

Yeasts are good candidates to utilize the hydrolysates of lignocellulose, the most abundant bioresource, for bioproducts. This study aimed to evaluate the efficiencies of single-cell protein (SCP) and xylitol production by a novel yeast strain, Candida intermedia FL023, from lignocellulosic hydrolysates and xylose. This strain efficiently assimilated hexose, pentose, and cellubiose for cell mass production with the crude protein content of 484.2 g kg -1 dry cell mass. SCP was produced by strain FL023 using corncob hydrolysate and urea as the carbon and nitrogen sources with the dry cell mass productivity 0.86 g L -1  h -1 and the yield of 0.40 g g -1 sugar. SCP was also produced using NaOH-pretreated Miscanthus sinensis straw and corn steep liquor as the carbon and nitrogen sources through simultaneous saccharification and fermentation with the dry cell productivity of 0.23 g L -1  h -1 and yield of 0.17 g g -1 straw. C. intermedia FL023 was tolerant to 0.5 g L -1 furfural, acetic acid, and syringaldehyde in xylitol fermentation and produced 45.7 g L -1 xylitol from xylose with the productivity of 0.38 g L -1  h -1 and the yield of 0.57 g g -1 xylose. This study provides feasible methods for feed and food additive production from the abundant lignocellulosic bioresources.

Isolation and characterization of acetate-utilizing anaerobes from a freshwater sediment

NARCIS (Netherlands)

Scholten, J.C.M.; Stams, A.J.M.

2000-01-01

Acetate-degrading anaerobic microorganisms in freshwater sediment were quantified by the most probable number technique. From the highest dilutions a methanogenic, a sulfate-reducing, and a nitrate-reducing microorganism were isolated with acetate as substrate. The methanogen (culture AMPB-Zg) was

Microalgae Cultivation on Anaerobic Digestate of Municipal Wastewater, Sewage Sludge and Agro-Waste

Directory of Open Access Journals (Sweden)

Luca Zuliani

2016-10-01

Full Text Available Microalgae are fast-growing photosynthetic organisms which have the potential to be exploited as an alternative source of liquid fuels to meet growing global energy demand. The cultivation of microalgae, however, still needs to be improved in order to reduce the cost of the biomass produced. Among the major costs encountered for algal cultivation are the costs for nutrients such as CO2, nitrogen and phosphorous. In this work, therefore, different microalgal strains were cultivated using as nutrient sources three different anaerobic digestates deriving from municipal wastewater, sewage sludge or agro-waste treatment plants. In particular, anaerobic digestates deriving from agro-waste or sewage sludge treatment induced a more than 300% increase in lipid production per volume in Chlorella vulgaris cultures grown in a closed photobioreactor, and a strong increase in carotenoid accumulation in different microalgae species. Conversely, a digestate originating from a pilot scale anaerobic upflow sludge blanket (UASB was used to increase biomass production when added to an artificial nutrient-supplemented medium. The results herein demonstrate the possibility of improving biomass accumulation or lipid production using different anaerobic digestates.

Selective Preparation of Furfural from Xylose over Sulfonic Acid Functionalized Mesoporous Sba-15 Materials

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Panpan Li

2011-04-01

Full Text Available Sulfonic acid functionalized mesoporous SBA-15 materials were prepared using the co-condensation and grafting methods, respectively, and their catalytic performance in the dehydration of xylose to furfural was examined. SBA-15-SO3H(C prepared by the co-condensation method showed 92–95% xylose conversion and 74% furfural selectivity, and 68–70% furfural yield under the given reaction conditions. The deactivation and regeneration of the SBA-15-SO3H(C catalyst for the dehydration of xylose was also investigated. The results indicate that the used and regeneration catalysts retained the SBA-15 mesoporous structure, and the S content of SBA-15-SO3H(C almost did not change. The deactivation of the catalysts is proposed to be associated with the accumulation of byproducts, which is caused by the loss reaction of furfural. After regeneration by H2O2, the catalytic activity of the catalyst almost recovered.

Evolutionary engineering of Saccharomyces cerevisiae for efficient aerobic xylose consumption

DEFF Research Database (Denmark)

Scalcinati, Gionata; Otero, José Manuel; Van Vleet, Jennifer R. H.

2012-01-01

Industrial biotechnology aims to develop robust microbial cell factories, such as Saccharomyces cerevisiae, to produce an array of added value chemicals presently dominated by petrochemical processes. Xylose is the second most abundant monosaccharide after glucose and the most prevalent pentose s...

Influence of genetic background of engineered xylose-fermenting industrial Saccharomyces cerevisiae strains for ethanol production from lignocellulosic hydrolysates

Science.gov (United States)

An industrial ethanol-producing Saccharomyces cerevisiae strain with genes needed for xylose-fermentation integrated into its genome was used to obtain haploids and diploid isogenic strains. The isogenic strains were more effective in metabolizing xylose than their parental strain (p < 0.05) and abl...

Antimicrobial Susceptibility of Enteric Gram Negative Facultative Anaerobe Bacilli in Aerobic versus Anaerobic Conditions

Science.gov (United States)

Amachawadi, Raghavendra G.; Renter, David G.; Volkova, Victoriya V.

2016-01-01

Antimicrobial treatments result in the host’s enteric bacteria being exposed to the antimicrobials. Pharmacodynamic models can describe how this exposure affects the enteric bacteria and their antimicrobial resistance. The models utilize measurements of bacterial antimicrobial susceptibility traditionally obtained in vitro in aerobic conditions. However, in vivo enteric bacteria are exposed to antimicrobials in anaerobic conditions of the lower intestine. Some of enteric bacteria of food animals are potential foodborne pathogens, e.g., Gram-negative bacilli Escherichia coli and Salmonella enterica. These are facultative anaerobes; their physiology and growth rates change in anaerobic conditions. We hypothesized that their antimicrobial susceptibility also changes, and evaluated differences in the susceptibility in aerobic vs. anaerobic conditions of generic E. coli and Salmonella enterica of diverse serovars isolated from cattle feces. Susceptibility of an isolate was evaluated as its minimum inhibitory concentration (MIC) measured by E-Test® following 24 hours of adaptation to the conditions on Mueller-Hinton agar, and on a more complex tryptic soy agar with 5% sheep blood (BAP) media. We considered all major antimicrobial drug classes used in the U.S. to treat cattle: β-lactams (specifically, ampicillin and ceftriaxone E-Test®), aminoglycosides (gentamicin and kanamycin), fluoroquinolones (enrofloxacin), classical macrolides (erythromycin), azalides (azithromycin), sulfanomides (sulfamethoxazole/trimethoprim), and tetracyclines (tetracycline). Statistical analyses were conducted for the isolates (n≥30) interpreted as susceptible to the antimicrobials based on the clinical breakpoint interpretation for human infection. Bacterial susceptibility to every antimicrobial tested was statistically significantly different in anaerobic vs. aerobic conditions on both media, except for no difference in susceptibility to ceftriaxone on BAP agar. A satellite experiment

Anaerobic oxidation of methane in grassland soils used for cattle husbandry

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A. Bannert

2012-10-01

Full Text Available While the importance of anaerobic methane oxidation has been reported for marine ecosystems, the role of this process in soils is still questionable. Grasslands used as pastures for cattle overwintering show an increase in anaerobic soil micro-sites caused by animal treading and excrement deposition. Therefore, anaerobic potential methane oxidation activity of severely impacted soil from a cattle winter pasture was investigated in an incubation experiment under anaerobic conditions using ¹³C-labelled methane. We were able to detect a high microbial activity utilizing CH₄ as nutrient source shown by the respiration of ¹³CO₂. Measurements of possible terminal electron acceptors for anaerobic oxidation of methane were carried out. Soil sulfate concentrations were too low to explain the oxidation of the amount of methane added, but enough nitrate and iron(III were detected. However, only nitrate was consumed during the experiment. ¹³C-PLFA analyses clearly showed the utilization of CH₄ as nutrient source mainly by organisms harbouring 16:1ω7 PLFAs. These lipids were also found as most ¹³C-enriched fatty acids by Raghoebarsing et al. (2006 after addition of ¹³CH₄ to an enrichment culture coupling denitrification of nitrate to anaerobic oxidation of methane. This might be an indication for anaerobic oxidation of methane by relatives of "Candidatus Methylomirabilis oxyfera" in the investigated grassland soil under the conditions of the incubation experiment.

Methylomusa anaerophila gen. nov., sp. nov., an anaerobic methanol-utilizing bacterium isolated from a microbial fuel cell.

Science.gov (United States)

Amano, Nanako; Yamamuro, Ayaka; Miyahara, Morio; Kouzuma, Atsushi; Abe, Takashi; Watanabe, Kazuya

2018-04-01

Abacterial strain, designated MMFC1 T , was isolated from a methanol-fed microbial fuel cell that had been inoculated with sludge obtained from a wastewater-treatmentfacility in a chemical plant. The strain grows by fermenting methanol to produce acetate under anaerobic conditions, while homoacetogenic growth is not observed. MMFC1 T also grows on pyruvate and lactate but not on sugars and other organic acids. Cells are curved rods and motile, have peritrichous flagella, and form endospores. The genome sequence of strain MMFC1 T supports the physiological data. Phylogenetic analysis based on the 16S rRNA gene sequence shows that strain MMFC1 T is affiliated with the family Sporomusaceae, while the closest relative is Sporomusa ovata with nucleotide-sequencesimilarity of 93.5 %. Major fatty acids are iso-C13 : 0 3-OH, C16 : 1ω9 and iso-C17 : 0. On the basis of its physiological, genomic and phylogenetic features, a novel genus and species are proposed to accommodate strain MMFC1 T , with the name Methylomusa anaerophila gen. nov., sp. nov. The type strain of Methylomusa anaerophila is MMFC1 T (=JCM 31821 T = KCTC 15592 T ).

Trends in the development of equipment for anaerobic fermentation

Energy Technology Data Exchange (ETDEWEB)

Holcak, B; Lutcha, J

1982-01-01

The successful application of anaerobic fermentation to the utilization of diluted wastes for the production of energy stimulated in recent years the development of new types of anaerobic reactors. Although the point of view of a chemical engineer does not encompass the complexity of this microbial process, he still disposes of means that enable him to estimate to what extent is it possible to affect the efficiency of the process by the concept of reactor arrangement. Simulation of behaviour by means of mathematical models enables us to compare quantitatively, for the types of anaerobic reactor under consideration, the apparatuses, and to predict the expected trends in their development.

The alcohol dehydrogenase system in the xylose-fermenting yeast Candida maltosa.

Directory of Open Access Journals (Sweden)

Yuping Lin

2010-07-01

Full Text Available The alcohol dehydrogenase (ADH system plays a critical role in sugar metabolism involving in not only ethanol formation and consumption but also the general "cofactor balance" mechanism. Candida maltosa is able to ferment glucose as well as xylose to produce a significant amount of ethanol. Here we report the ADH system in C. maltosa composed of three microbial group I ADH genes (CmADH1, CmADH2A and CmADH2B, mainly focusing on its metabolic regulation and physiological function.Genetic analysis indicated that CmADH2A and CmADH2B tandemly located on the chromosome could be derived from tandem gene duplication. In vitro characterization of enzymatic properties revealed that all the three CmADHs had broad substrate specificities. Homo- and heterotetramers of CmADH1 and CmADH2A were demonstrated by zymogram analysis, and their expression profiles and physiological functions were different with respect to carbon sources and growth phases. Fermentation studies of ADH2A-deficient mutant showed that CmADH2A was directly related to NAD regeneration during xylose metabolism since CmADH2A deficiency resulted in a significant accumulation of glycerol.Our results revealed that CmADH1 was responsible for ethanol formation during glucose metabolism, whereas CmADH2A was glucose-repressed and functioned to convert the accumulated ethanol to acetaldehyde. To our knowledge, this is the first demonstration of function separation and glucose repression of ADH genes in xylose-fermenting yeasts. On the other hand, CmADH1 and CmADH2A were both involved in ethanol formation with NAD regeneration to maintain NADH/NAD ratio in favor of producing xylitol from xylose. In contrast, CmADH2B was expressed at a much lower level than the other two CmADH genes, and its function is to be further confirmed.

Identification of a novel acetate-utilizing bacterium belonging to Synergistes group 4 in anaerobic digester sludge.

Science.gov (United States)

Ito, Tsukasa; Yoshiguchi, Kazumi; Ariesyady, Herto Dwi; Okabe, Satoshi

2011-12-01

Major acetate-utilizing bacterial and archaeal populations in methanogenic anaerobic digester sludge were identified and quantified by radioisotope- and stable-isotope-based functional analyses, microautoradiography-fluorescence in situ hybridization (MAR-FISH) and stable-isotope probing of 16S rRNA (RNA-SIP) that can directly link 16S rRNA phylogeny with in situ metabolic function. First, MAR-FISH with (14)C-acetate indicated the significant utilization of acetate by only two major groups, unidentified bacterial cells and Methanosaeta-like filamentous archaeal cells, in the digester sludge. To identify the acetate-utilizing unidentified bacteria, RNA-SIP was conducted with (13)C(6)-glucose and (13)C(3)-propionate as sole carbon source, which were followed by phylogenetic analysis of 16S rRNA. We found that bacteria belonging to Synergistes group 4 were commonly detected in both 16S rRNA clone libraries derived from the sludge incubated with (13)C-glucose and (13)C-propionate. To confirm that this bacterial group can utilize acetate, specific FISH probe targeting for Synergistes group 4 was newly designed and applied to the sludge incubated with (14)C-acetate for MAR-FISH. The MAR-FISH result showed that bacteria belonging to Synergistes group 4 significantly took up acetate and their active population size was comparable to that of Methanosaeta in this sludge. In addition, as bacteria belonging to Synergistes group 4 had high K(m) for acetate and maximum utilization rate, they are more competitive for acetate over Methanosaeta at high acetate concentrations (2.5-10  mM). To our knowledge, it is the first time to report the acetate-utilizing activity of uncultured bacteria belonging to Synergistes group 4 and its competitive significance to acetoclastic methanogen, Methanosaeta.

Identification and characterization of D-xylulokinase from the D-xylose-fermenting fungus, Mucor circinelloides.

Science.gov (United States)

Komeda, Hidenobu; Yamasaki-Yashiki, Shino; Hoshino, Kazuhiro; Asano, Yasuhisa

2014-11-01

D-Xylulokinase catalyzes the phosphorylation of D-xylulose in the final step of the pentose catabolic pathway to form d-xylulose-5-phosphate. The D-xylulokinase activity was found to be induced by both D-xylose and L-arabinose, as well as some of the other enzymes involved in the pentose catabolism, in the D-xylose-fermenting zygomycetous fungus, Mucor circinelloides NBRC 4572. The putative gene, xyl3, which may encode D-xylulokinase, was detected in the genome sequence of this strain. The amino acid sequence deduced from the gene was more similar to D-xylulokinases from an animal origin than from other fungi. The recombinant enzyme was purified from the E. coli transformant expressing xyl3 and then characterized. The ATP-dependent phosphorylative activity of the enzyme was the highest toward D-xylulose. Its kinetic parameters were determined as Km (D-xylulose) = 0.29 mM and Km (ATP) = 0.51 mM, indicating that the xyl3 gene encoded D-xylulokinase (McXK). Western blot analysis revealed that McXK was induced by L-arabinose as well as D-xylose and the induction was repressed in the presence of D-glucose, suggesting that the enzyme may be involved in the catabolism of D-xylose and L-arabinose and is subject to carbon catabolite repression in this fungus. This is the first study on D-xylulokinase from zygomycetous fungi. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

Development of technology for fuel alcohol. ; Searching and breeding of superior bacteria. Nenryoyo alcohol gijutsu kaihatsu. ; Yushukin no kensakuter dot ikushu

Energy Technology Data Exchange (ETDEWEB)

1990-10-01

In order to develop superior ethanol fermenting bacteria which would directly transform fibers into ethanol, which would lead to the utilization of unused waste industrial agricultural and forestry products, separation of potential strains from extensive collections of microbe resources from in and out of this country and adding of salt resistance, alcohol resistance, cellulose digestion properties using bio-technology were conducted. In searching for superior bacteria, some cells used in the sugar test plant showed no decrease in fermentation yields in 0.4% CaCl{sub 2}, 0.25 MgCl{sub 2} cultures, and some showed no decrease even with 1% KCl. In the breeding of superior bacteria, zymomonus with 4 times CMCase (a component enzyme of a cellulose degrading enzyme) activity, zymomonus which could grow in maltose cultures, zymomonus which had xylose isomerase genes linked to pyruvate carboxylase promoters, zymomonus resistant to 7% ethanol and 1% KCl, thermophilic and anaerobic cellulose degrading bacteria were developed. 1 tab.

Increased accuracy of the carbon-14 D-xylose breath test in detecting small-intestinal bacterial overgrowth by correction with the gastric emptying rate

International Nuclear Information System (INIS)

Chang Chisen; Chen Granhum; Kao Chiahung; Wang Shyhjen; Peng Shihnen; Huang Chihkuen; Poon Sekkwong

1995-01-01

The aim of this study was to determine whether the accuracy of 14 C-D-xylose breath test for detecting bacterial overgrowth can be increased by correction with the gastric emptying rate of 14 C-D-xylose. Ten culture-positive patients and ten culture-negative controls were included in the study. Small-intestinal aspirates for bacteriological culture were obtained endoscopically. A liquid-phase gastric emptying study was performed simultaneously to assess the amount of 14 C-D-xylose that entered the small intestine. The results of the percentage of expired 14 CO 2 at 30 min were corrected with the amount of 14 C-D-xylose that entered the small intestine. There were six patients in the culture-positive group with a 14 CO 2 concentration above the normal limit. Three out of four patients with initially negative results using the uncorrected method proved to be positive after correction. All these three patients had prolonged gastric emptying of 14 C-D-xylose. When compared with cultures of small-intestine aspirates, the sensitivity and specificity of the uncorrected 14 C-D-xylose breath test were 60% and 90%, respectively. In contrast, the sensitivity and specificity of the corrected 14 C-D-xylose breath test improved to 90% and 100%, respectively. (orig./MG)

Improved bioethanol production using fusants of Saccharomyces cerevisiae and xylose-fermenting yeasts.

Science.gov (United States)

Kumari, Rajni; Pramanik, K

2012-06-01

The present research deals with the development of a hybrid yeast strain with the aim of converting pentose and hexose sugar components of lignocellulosic substrate to bioethanol by fermentation. Different fusant strains were obtained by fusing protoplasts of Saccharomyces cerevisiae and xylose-fermenting yeasts such as Pachysolen tannophilus, Candida shehatae and Pichia stipitis. The fusants were sorted by fluorescent-activated cell sorter and further confirmed by molecular characterization. The fusants were evaluated by fermentation of glucose-xylose mixture and the highest ethanol producing fusant was used for further study to ferment hydrolysates produced by acid pretreatment and enzymatic hydrolysis of cotton gin waste. Among the various fusant and parental strains used under present study, RPR39 was found to be stable and most efficient strain giving maximum ethanol concentration (76.8 ± 0.31 g L(-1)), ethanol productivity (1.06 g L(-1) h(-1)) and ethanol yield (0.458 g g(-1)) by fermentation of glucose-xylose mixture under test conditions. The fusant has also shown encouraging result in fermenting hydrolysates of cotton gin waste with ethanol concentration of 7.08 ± 0.142 g L(-1), ethanol yield of 0.44 g g(-1), productivity of 0.45 g L(-1) h(-1) and biomass yield of 0.40 g g(-1).

The fate of methanol in anaerobic bioreactors

NARCIS (Netherlands)

Florencio, L.

1994-01-01

Methanol is an important component of certain industrial wastewaters. In anaerobic environments, methanol can be utilized by methanogens and acetogens. In wastewater treatment plants, the conversion of methanol into methane is preferred because this conversion is responsible for chemical

Evolutionary engineering of Saccharomyces cerevisiae for efficient aerobic xylose consumption

Science.gov (United States)

Gionata Scalcinati; Jose´ Manuel Otero; Jennifer R.H. Van Vleet; Thomas W. Jeffries; Lisbeth Olsson; Jens. Nielsen

2012-01-01

Industrial biotechnology aims to develop robust microbial cell factories, such as , to produce an array of added value chemicals presently dominated by petrochemical processes. Xylose is the second most abundant monosaccharide after glucose and the most prevalent pentose sugar found in lignocelluloses. Significant research...

ANAEROBIC DEGRADATION OF HALOGENATED BENZOIC-ACIDS BY PHOTOHETEROTROPHIC BACTERIA

NARCIS (Netherlands)

VANDERWOUDE, BJ; DEBOER, M; VANDERPUT, NMJ; VANDERGELD, FM; PRINS, RA; GOTTSCHAL, JC

1994-01-01

From light-exposed enrichment cultures containing benzoate and a mixture of chlorobenzoates, a pure culture was obtained able to grow with 3-chlorobenzoate (3-CBA) or 3-bromobenzoate (3-BrBA) as the sole growth substrate anaerobically in the light. The thus isolated organism is a photoheterotroph,

An innovative biocatalyst for production of ethanol from xylose in a continuous bioreactor.

Science.gov (United States)

Silva, C R; Zangirolami, T C; Rodrigues, J P; Matugi, K; Giordano, R C; Giordano, R L C

2012-01-05

The use of the hemicellulose fraction of biomass may be important for the feasibility of the production of second generation bioethanol. Wild strains of Saccharomyces cerevisiae are widely used in industry for production of 1st generation ethanol, and the robustness of this yeast is an important advantage in large scale applications. Isomerization of xylose to xylulose is an essential step in this process. This reaction is catalyzed by glucose isomerase (GI). A new biocatalyst is presented here for the simultaneous isomerization and fermentation (SIF) of xylose. GI from Streptomyces rubiginosus was immobilized in chitosan, through crosslinking with glutaraldehyde, and the support containing the immobilized GI (IGI-Ch) was co-immobilized with S. cerevisiae, in calcium alginate gel. The immobilization experiments led to high immobilized protein loads (30-68 mg × g(support)(-1)), high yields (circa of 100%) and high recovered enzyme activity (>90%). The IGI-Ch derivative with maximum activity presented 1700 IU × g(catalyst)(-1), almost twice the activity of a commercial immobilized GI, GENSWEET(®) IGI-HF. At typical operational conditions for xylose SIF operation (pH 5, 30-35 °C, presence of nutrients and ethanol concentrations in the medium up to 70 L(-1)), both derivatives, IGI-Ch and GENSWEET(®) IGI-HF retained app. 90% of the initial activity after 120 h, while soluble GI was almost completely inactive at pH 5, 30 °C. The isomerization xylose/xylulose, catalyzed by IGI-Ch, reached the equilibrium in batch experiments after 4h, with 12,000 IU × L(-1) (7 g(der) × L(-1)), at pH 5 and 30 °C, in the presence of fermentation nutrients. After co-immobilization of IGI-Ch with yeast in alginate gel, this biocatalyst succeeded in producing 12 g × L(-1) of ethanol, 9.5 g × L(-1) of xylitol, 2.5 g × L(-1) of glycerol and 1.9 g × L(-1) of acetate after consumption of 50 g × L(-1) of xylose, in 48 h, using 32.5 × 10(3) IU × L(-1) and 20 g(yeast) × L(-1), at 35

Caldicellulosiruptor obsidiansis sp. nov., an anaerobic, extremely thermophilic, cellulolytic bacterium isolated from Obsidian Pool, Yellowstone National Park.

Science.gov (United States)

Hamilton-Brehm, Scott D; Mosher, Jennifer J; Vishnivetskaya, Tatiana; Podar, Mircea; Carroll, Sue; Allman, Steve; Phelps, Tommy J; Keller, Martin; Elkins, James G

2010-02-01

A novel, obligately anaerobic, extremely thermophilic, cellulolytic bacterium, designated OB47(T), was isolated from Obsidian Pool, Yellowstone National Park, WY. The isolate was a nonmotile, non-spore-forming, Gram-positive rod approximately 2 microm long by 0.2 microm wide and grew at temperatures between 55 and 85 degrees C, with the optimum at 78 degrees C. The pH range for growth was 6.0 to 8.0, with values of near 7.0 being optimal. Growth on cellobiose produced the fastest specific growth rate at 0.75 h(-1). The organism also displayed fermentative growth on glucose, maltose, arabinose, fructose, starch, lactose, mannose, sucrose, galactose, xylose, arabinogalactan, Avicel, xylan, filter paper, processed cardboard, pectin, dilute acid-pretreated switchgrass, and Populus. OB47(T) was unable to grow on mannitol, fucose, lignin, Gelrite, acetate, glycerol, ribose, sorbitol, carboxymethylcellulose, and casein. Yeast extract stimulated growth, and thiosulfate, sulfate, nitrate, and sulfur were not reduced. Fermentation end products were mainly acetate, H2, and CO2, although lactate and ethanol were produced in 5-liter batch fermentations. The G+C content of the DNA was 35 mol%, and sequence analysis of the small subunit rRNA gene placed OB47(T) within the genus Caldicellulosiruptor. Based on its phylogenetic and phenotypic properties, the isolate is proposed to be designated Caldicellulosiruptor obsidiansis sp. nov. and OB47 is the type strain (ATCC BAA-2073).

Caldicellulosiruptor obsidiansis sp. nov., an Anaerobic, Extremely Thermophilic, Cellulolytic Bacterium Isolated from Obsidian Pool, Yellowstone National Park▿

Science.gov (United States)

Hamilton-Brehm, Scott D.; Mosher, Jennifer J.; Vishnivetskaya, Tatiana; Podar, Mircea; Carroll, Sue; Allman, Steve; Phelps, Tommy J.; Keller, Martin; Elkins, James G.

2010-01-01

A novel, obligately anaerobic, extremely thermophilic, cellulolytic bacterium, designated OB47T, was isolated from Obsidian Pool, Yellowstone National Park, WY. The isolate was a nonmotile, non-spore-forming, Gram-positive rod approximately 2 μm long by 0.2 μm wide and grew at temperatures between 55 and 85°C, with the optimum at 78°C. The pH range for growth was 6.0 to 8.0, with values of near 7.0 being optimal. Growth on cellobiose produced the fastest specific growth rate at 0.75 h−1. The organism also displayed fermentative growth on glucose, maltose, arabinose, fructose, starch, lactose, mannose, sucrose, galactose, xylose, arabinogalactan, Avicel, xylan, filter paper, processed cardboard, pectin, dilute acid-pretreated switchgrass, and Populus. OB47T was unable to grow on mannitol, fucose, lignin, Gelrite, acetate, glycerol, ribose, sorbitol, carboxymethylcellulose, and casein. Yeast extract stimulated growth, and thiosulfate, sulfate, nitrate, and sulfur were not reduced. Fermentation end products were mainly acetate, H2, and CO2, although lactate and ethanol were produced in 5-liter batch fermentations. The G+C content of the DNA was 35 mol%, and sequence analysis of the small subunit rRNA gene placed OB47T within the genus Caldicellulosiruptor. Based on its phylogenetic and phenotypic properties, the isolate is proposed to be designated Caldicellulosiruptor obsidiansis sp. nov. and OB47 is the type strain (ATCC BAA-2073). PMID:20023107

Development of industrial yeast for second generation bioethanol production

Energy Technology Data Exchange (ETDEWEB)

Hou, X

2012-01-15

The cost of lignocellulose-based bioethanol needs to be reduced, in order to commercialize this clean and sustainable fuel substitute for fossil fuels. A microorganism that can completely and efficiently convert all the sugars in lignocellulose into ethanol is one of the prerequisites of a cost-effective production process. In addition, the microorganisms should also have a high tolerance towards the inhibitory compounds present in the lignocellulosic hydrolysate, which are formed during the pretreatment of lignocellulose. Baker's yeast, Saccharomyces cerevisiae, is generally regarded as a robust microorganism and can efficiently ferment glucose. But it lacks the ability to ferment xylose which comprises 20-35% of lignocellulose. Naturally xylose-fermenting yeast such as Pichia stipitis is much more sensitive to inhibitors than S. cerevisiae and it requires accurately controlled microaerophilic conditions during the xylose fermentation, rendering the process technically difficult and expensive. In this study, a novel xylose fermenting yeast Spathaspora passalidarum displayed fast cell growth and efficient xylose fermentation under anaerobic conditions. In contrast, P. stipitis was almost unable to utilize xylose under the same conditions. It is further demonstrated that S. passalidarum converts xylose by means of NADH-preferred xylose reductase (XR) and NAD+-dependent xylitol dehydrogenase (XDH). Thus, the capacity of S. passalidarum to utilize xylose under anaerobic conditions is possibly due to a balance between supply and demand of cofactor through this XR-XDH pathway. Only one other XR with NADH preference has been reported so far. Unfortunately, S. passalidarum also has a low tolerance towards inhibitors generated during pretreatment, which prevents immediate use of this yeast in industrial application. S. passalidarum is able to convert the inhibitor furfural to furfuryl alcohol in a synthetic medium when the addition of furfural is low. The enzymes involved

Development of industrial yeast for second generation bioethanol production

Energy Technology Data Exchange (ETDEWEB)

Hou, X.

2012-01-15

The cost of lignocellulose-based bioethanol needs to be reduced, in order to commercialize this clean and sustainable fuel substitute for fossil fuels. A microorganism that can completely and efficiently convert all the sugars in lignocellulose into ethanol is one of the prerequisites of a cost-effective production process. In addition, the microorganisms should also have a high tolerance towards the inhibitory compounds present in the lignocellulosic hydrolysate, which are formed during the pretreatment of lignocellulose. Baker's yeast, Saccharomyces cerevisiae, is generally regarded as a robust microorganism and can efficiently ferment glucose. But it lacks the ability to ferment xylose which comprises 20-35% of lignocellulose. Naturally xylose-fermenting yeast such as Pichia stipitis is much more sensitive to inhibitors than S. cerevisiae and it requires accurately controlled microaerophilic conditions during the xylose fermentation, rendering the process technically difficult and expensive. In this study, a novel xylose fermenting yeast Spathaspora passalidarum displayed fast cell growth and efficient xylose fermentation under anaerobic conditions. In contrast, P. stipitis was almost unable to utilize xylose under the same conditions. It is further demonstrated that S. passalidarum converts xylose by means of NADH-preferred xylose reductase (XR) and NAD+-dependent xylitol dehydrogenase (XDH). Thus, the capacity of S. passalidarum to utilize xylose under anaerobic conditions is possibly due to a balance between supply and demand of cofactor through this XR-XDH pathway. Only one other XR with NADH preference has been reported so far. Unfortunately, S. passalidarum also has a low tolerance towards inhibitors generated during pretreatment, which prevents immediate use of this yeast in industrial application. S. passalidarum is able to convert the inhibitor furfural to furfuryl alcohol in a synthetic medium when the addition of furfural is low. The enzymes

Evaluation of marine algae as a source of biogas in a two-stage anaerobic reactor system

International Nuclear Information System (INIS)

Vergara-Fernandez, Alberto; Vargas, Gisela; Alarcon, Nelson; Velasco, Antonio

2008-01-01

The marine algae are considered an important biomass source; however, their utilization as energy source is still low around the world. The technical feasibility of marine algae utilization as a source of renewable energy was studied to laboratory scale. The anaerobic digestion of Macrocystis pyrifera, Durvillea antarctica and their blend 1:1 (w/w) was evaluated in a two-phase anaerobic digestion system, which consisted of an anaerobic sequencing batch reactor (ASBR) and an upflow anaerobic filter (UAF). The results show that 70% of the total biogas produced in the system was generated in the UAF, and both algae species have similar biogas productions of 180.4(±1.5) mL g -1 dry algae d -1 , with a methane concentration around 65%. The same methane content was observed in biogas yield of algae blend; however, a lower biogas yield was obtained. In conclusion, either algae species or their blend can be utilized to produce methane gas in a two-phase digestion system

Evaluation of marine algae as a source of biogas in a two-stage anaerobic reactor system

Energy Technology Data Exchange (ETDEWEB)

Vergara-Fernandez, Alberto; Vargas, Gisela [Escuela de Ingenieria Ambiental, Facultad de Ingenieria, Universidad Catolica de Temuco, Manuel Montt 56, Casilla 15-D, Temuco (Chile); Alarcon, Nelson [Departamento de Ingenieria Quimica, Facultad de Ingenieria y Ciencias Geologicas, Universidad Catolica del Norte (Chile); Velasco, Antonio [Centro Nacional de Investigacion y Capacitacion Ambiental del Instituto Nacional de Ecologia (CENICA-INE), Av. San Rafael Atlixco 186, Col. Vicentina, Del. Iztapalapa, 09340, Mexico, DF (Mexico)

2008-04-15

The marine algae are considered an important biomass source; however, their utilization as energy source is still low around the world. The technical feasibility of marine algae utilization as a source of renewable energy was studied to laboratory scale. The anaerobic digestion of Macrocystis pyrifera, Durvillea antarctica and their blend 1:1 (w/w) was evaluated in a two-phase anaerobic digestion system, which consisted of an anaerobic sequencing batch reactor (ASBR) and an upflow anaerobic filter (UAF). The results show that 70% of the total biogas produced in the system was generated in the UAF, and both algae species have similar biogas productions of 180.4({+-}1.5) mL g{sup -1} dry algae d{sup -1}, with a methane concentration around 65%. The same methane content was observed in biogas yield of algae blend; however, a lower biogas yield was obtained. In conclusion, either algae species or their blend can be utilized to produce methane gas in a two-phase digestion system. (author)

Data for rapid ethanol production at elevated temperatures by engineered thermotolerant Kluyveromyces marxianus via the NADP(H-preferring xylose reductase–xylitol dehydrogenase pathway

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Biao Zhang

2015-12-01

Full Text Available A thermo-tolerant NADP(H-preferring xylose pathway was constructed in Kluyveromyces marxianus for ethanol production with xylose at elevated temperatures (Zhang et al., 2015 [25]. Ethanol production yield and efficiency was enhanced by pathway engineering in the engineered strains. The constructed strain, YZJ088, has the ability to co-ferment glucose and xylose for ethanol and xylitol production, which is a critical step toward enabling economic biofuel production from lignocellulosic biomass. This study contains the fermentation results of strains using the metabolic pathway engineering procedure. The ethanol-producing abilities of various yeast strains under various conditions were compared, and strain YZJ088 showed the highest production and fastest productivity at elevated temperatures. The YZJ088 xylose fermentation results indicate that it fermented well with xylose at either low or high inoculum size. When fermented with an initial cell concentration of OD600=15 at 37 °C, YZJ088 consumed 200 g/L xylose and produced 60.07 g/L ethanol; when the initial cell concentration was OD600=1 at 37 °C, YZJ088 consumed 98.96 g/L xylose and produced 33.55 g/L ethanol with a productivity of 0.47 g/L/h. When fermented with 100 g/L xylose at 42 °C, YZJ088 produced 30.99 g/L ethanol with a productivity of 0.65 g/L/h, which was higher than that produced at 37 °C.

Importance of cobalt for individual trophic groups in an anaerobic methanol-degrading consortium.

OpenAIRE

Florencio, L; Field, J A; Lettinga, G

1994-01-01

Methanol is an important anaerobic substrate in industrial wastewater treatment and the natural environment. Previous studies indicate that cobalt greatly stimulates methane formation during anaerobic treatment of methanolic wastewaters. To evaluate the effect of cobalt in a mixed culture, a sludge with low background levels of cobalt was cultivated in an upflow anaerobic sludge blanket reactor. Specific inhibitors in batch assays were then utilized to study the effect of cobalt on the growth...

Comparative genomics of xylose-fermenting fungi for enhanced biofuel production

Science.gov (United States)

Dana J. Wolbach; Alan Kuo; Trey K. Sato; Katlyn M. Potts; Asaf A. Salamov; Kurt M. LaButti; Hui Sun; Alicia Clum; Jasmyn L. Pangilinan; Erika A. Lindquist; Susan Lucas; Alla Lapidus; Mingjie Jin; Christa Gunawan; Venkatesh Balan; Bruce E. Dale; Thomas W. Jeffries; Robert Zinkel; Kerrie W. Barry; Igor V. Grigoriev; Audrey P. Gasch

2011-01-01

Cellulosic biomass is an abundant and underused substrate for biofuel production. The inability of many microbes to metabolize the pentose sugars abundant within hemicellulose creates specific challenges for microbial biofuel production from cellulosic material. Although engineered strains of Saccharomyces cerevisiae can use the pentose xylose, the fermentative...

Some unique features of alkaliphilic anaerobes

Science.gov (United States)

Roof, Erin; Pikuta, Elena; Otto, Christopher; Williams, George; Hoover, Richard

2013-09-01

This article explores two topics involving the examination of four strains of alkaliphilic anaerobes. The first topic was dedicated to detection of the ability of microorganisms to metabolize alternative chirality substrates. Two saccharolytic anaerobic bacteria were chosen for the first experiment: Anaerovirgula multivorans strain SCAT, which is gram positive and spore-forming; and Spirochaeta dissipatitropha, strain ASpC2T, which is gram negative. It was found that both checked sugarlytics were able to use L-ribose and L-arabinose, as growth substrates. The second part was concerned of study a chemolithotrophy in two halo-alkaliphilic sulfate reducing bacteria: Desulfonatornum thiodismutans strain MLF1T and Desulfonatronum lacustre strain Z-7951T. The experiments with lithotrophs had demonstrated that strain MLF1T was capable to grow without any organic source of carbon, while strain Z-7951T had required at least 2 mM sodium acetate for growth. Anaerobic technique was used for preparation of the growth media and maintenance of these bacterial cultures. Standard methods for Gram, spore, and flagella staining were applied for characterization of cytomorphology. In this article, the results of the experiments performed on cytological, physiological, and biochemical levels are presented and discussed.

Design and Fabrication of an Anaerobic Digester

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M. S. Abubakar

2017-02-01

Full Text Available Anaerobic digester is a physical structure that provides a conducive environment for the multiplication of micro-organisms that degrades organic matter to generate biogas energy. Energy is required in agriculture for crop production, processing and storage, poultry production and electricity for farmstead and farm settlements. It is energy that propels agricultural mechanization, which minimizes the use of human and animal muscles and its inherent drudgery in agriculture. The energy demand required to meet up with the agricultural growth in Nigeria is high and growing every year. In this study the design and fabrication of an anaerobic digester was reported which is an attempt to boost energy requirement for small and medium dryland farmers in Nigeria. The design of the digester includes the following concept; the basic principles of anaerobic digestion processes, socio-economic status of the dryland farmers, amount of biogas to be produced. Finally, the digester was fabricated using locally available raw materials within the dryland area of Nigeria. At the end, preliminary flammability test was conducted and the biogas produced was found to be flammable.

Production of medium-chain-length polyhydroxyalkanoates by sequential feeding of xylose and octanoic acid in engineered Pseudomonas putida KT2440

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Le Meur Sylvaine

2012-08-01

Full Text Available Abstract Background Pseudomonas putida KT2440 is able to synthesize large amounts of medium-chain-length polyhydroxyalkanoates (mcl-PHAs. To reduce the substrate cost, which represents nearly 50% of the total PHA production cost, xylose, a hemicellulose derivate, was tested as the growth carbon source in an engineered P. putida KT2440 strain. Results The genes encoding xylose isomerase (XylA and xylulokinase (XylB from Escherichia coli W3110 were introduced into P. putida KT2440. The recombinant KT2440 exhibited a XylA activity of 1.47 U and a XylB activity of 0.97 U when grown on a defined medium supplemented with xylose. The cells reached a maximum specific growth rate of 0.24 h-1 and a final cell dry weight (CDW of 2.5 g L-1 with a maximal yield of 0.5 g CDW g-1 xylose. Since no mcl-PHA was accumulated from xylose, mcl-PHA production can be controlled by the addition of fatty acids leading to tailor-made PHA compositions. Sequential feeding strategy was applied using xylose as the growth substrate and octanoic acid as the precursor for mcl-PHA production. In this way, up to 20% w w-1 of mcl-PHA was obtained. A yield of 0.37 g mcl-PHA per g octanoic acid was achieved under the employed conditions. Conclusions Sequential feeding of relatively cheap carbohydrates and expensive fatty acids is a practical way to achieve more cost-effective mcl-PHA production. This study is the first reported attempt to produce mcl-PHA by using xylose as the growth substrate. Further process optimizations to achieve higher cell density and higher productivity of mcl-PHA should be investigated. These scientific exercises will undoubtedly contribute to the economic feasibility of mcl-PHA production from renewable feedstock.

Anaerobes as Sources of Bioactive Compounds and Health Promoting Tools.

Science.gov (United States)

Mamo, Gashaw

Aerobic microorganisms have been sources of medicinal agents for several decades and an impressive variety of drugs have been isolated from their cultures, studied and formulated to treat or prevent diseases. On the other hand, anaerobes, which are believed to be the oldest life forms on earth and evolved remarkably diverse physiological functions, have largely been neglected as sources of bioactive compounds. However, results obtained from the limited research done so far show that anaerobes are capable of producing a range of interesting bioactive compounds that can promote human health. In fact, some of these bioactive compounds are found to be novel in their structure and/or mode of action.Anaerobes play health-promoting roles through their bioactive products as well as application of whole cells. The bioactive compounds produced by these microorganisms include antimicrobial agents and substances such as immunomodulators and vitamins. Bacteriocins produced by anaerobes have been in use as preservatives for about 40 years. Because these substances are effective at low concentrations, encounter relatively less resistance from bacteria and are safe to use, there is a growing interest in these antimicrobial agents. Moreover, several antibiotics have been reported from the cultures of anaerobes. Closthioamide and andrimid produced by Clostridium cellulolyticum and Pantoea agglomerans, respectively, are examples of novel antibiotics of anaerobe origin. The discovery of such novel bioactive compounds is expected to encourage further studies which can potentially lead to tapping of the antibiotic production potential of this fascinating group of microorganisms.Anaerobes are widely used in preparation of fermented foods and beverages. During the fermentation processes, these organisms produce a number of bioactive compounds including anticancer, antihypertensive and antioxidant substances. The well-known health promoting effect of fermented food is mostly due to these

Low acid hydrothermal fractionation of Giant Miscanthus for production of xylose-rich hydrolysate and furfural.

Science.gov (United States)

Kim, Tae Hyun; Ryu, Hyun Jin; Oh, Kyeong Keun

2016-10-01

Low acid hydrothermal (LAH) fractionation was developed for the effective recovery of hemicellulosic sugar (mainly xylose) from Miscanthus sacchariflorus Goedae-Uksae 1 (M. GU-1). The xylose yield was maximized at 74.75% when the M. GU-1 was fractionated at 180°C and 0.3wt.% of sulfuric acid for 10min. At this condition, the hemicellulose (mainly xylan) degradation was 86.41%. The difference between xylan degradation and xylose recovery yield, i.e., xylan loss, was 11.66%, as indicated by the formation of decomposed products. The furfural, the value added biochemical product, was also obtained by 0.42g/L at this condition, which was 53.82% of furfural production yield based on the xylan loss. After then, the furfural production continued to increase to a maximum concentration of 1.87g/L, at which point the xylan loss corresponded to 25.87%. Copyright © 2016 Elsevier Ltd. All rights reserved.

Diversity and ubiquity of bacteria capable of utilizing humic substances as electron donors for anaerobic respiration.

Science.gov (United States)

Coates, John D; Cole, Kimberly A; Chakraborty, Romy; O'Connor, Susan M; Achenbach, Laurie A

2002-05-01

Previous studies have demonstrated that reduced humic substances (HS) can be reoxidized by anaerobic bacteria such as Geobacter, Geothrix, and Wolinella species with a suitable electron acceptor; however, little is known of the importance of this metabolism in the environment. Recently we investigated this metabolism in a diversity of environments including marine and aquatic sediments, forest soils, and drainage ditch soils. Most-probable-number enumeration studies were performed using 2,6-anthrahydroquinone disulfonate (AHDS), an analog for reduced HS, as the electron donor with nitrate as the electron acceptor. Anaerobic organisms capable of utilizing reduced HS as an electron donor were found in all environments tested and ranged from a low of 2.31 x 10(1) in aquifer sediments to a high of 9.33 x 10(6) in lake sediments. As part of this study we isolated six novel organisms capable of anaerobic AHDS oxidation. All of the isolates coupled the oxidation of AHDS to the reduction of nitrate with acetate (0.1 mM) as the carbon source. In the absence of cells, no AHDS oxidation was apparent, and in the absence of AHDS, no cell density increase was observed. Generally, nitrate was reduced to N(2). Analysis of the AHDS and its oxidized form, 2,6-anthraquinone disulfonate (AQDS), in the medium during growth revealed that the anthraquinone was not being biodegraded as a carbon source and was simply being oxidized as an energy source. Determination of the AHDS oxidized and nitrate reduced accounted for 109% of the theoretical electron transfer. In addition to AHDS, all of these isolates could also couple the oxidation of reduced humic substances to the reduction of nitrate. No HS oxidation occurred in the absence of cells and in the absence of a suitable electron acceptor, demonstrating that these organisms were capable of utilizing natural HS as an energy source and that AHDS serves as a suitable analog for studying this metabolism. Alternative electron donors included

Urea utilization in growing lambs. 7

International Nuclear Information System (INIS)

Ulbrich, M.

1989-01-01

The utilization quota of NPN and pure feed protein for body protein synthesis was calculated on the basis of N balance experiments with 15 N-labelled urea with the help of model concepts of a 3-pool model and its mathematical usage. In lambs weighing 13 kg the efficiency of amino acid and nucleic acid synthesis in the non-amino acid N pool was 64%. This results in a total utilization quota for NPN and pure protein in the ration of 40% and 60%, resp. Lambs weighing 27 kg showed an efficiency in amino acid and nucleic acid synthesis in the non-AA N pool of 77% and in the AA N pool of 60%. The total utilization quota of NPN was 47% and that of pure protein 56%. The pure protein in the ration was thus about twice as well utilized for total protein synthesis and for protein syntesis for crude protein retention as the NPN compounds in the ration. (author)

Overexpression of pyruvate decarboxylase in the yeast Hansenula polymorpha results in increased ethanol yield in high-temperature fermentation of xylose.

Science.gov (United States)

Ishchuk, Olena P; Voronovsky, Andriy Y; Stasyk, Oleh V; Gayda, Galina Z; Gonchar, Mykhailo V; Abbas, Charles A; Sibirny, Andriy A

2008-11-01

Improvement of xylose fermentation is of great importance to the fuel ethanol industry. The nonconventional thermotolerant yeast Hansenula polymorpha naturally ferments xylose to ethanol at high temperatures (48-50 degrees C). Introduction of a mutation that impairs ethanol reutilization in H. polymorpha led to an increase in ethanol yield from xylose. The native and heterologous (Kluyveromyces lactis) PDC1 genes coding for pyruvate decarboxylase were expressed at high levels in H. polymorpha under the control of the strong constitutive promoter of the glyceraldehyde-3-phosphate dehydrogenase gene (GAPDH). This resulted in increased pyruvate decarboxylase activity and improved ethanol production from xylose. The introduction of multiple copies of the H. polymorpha PDC1 gene driven by the strong constitutive promoter led to a 20-fold increase in pyruvate decarboxylase activity and up to a threefold elevation of ethanol production.

A Review of the Anaerobic Digestion of Fruit and Vegetable Waste.

Science.gov (United States)

Ji, Chao; Kong, Chui-Xue; Mei, Zi-Li; Li, Jiang

2017-11-01

Fruit and vegetable waste is an ever-growing global question. Anaerobic digestion techniques have been developed that facilitate turning such waste into possible sources for energy and fertilizer, simultaneously helping to reduce environmental pollution. However, various problems are encountered in applying these techniques. The purpose of this study is to review local and overseas studies, which focus on the use of anaerobic digestion to dispose fruit and vegetable wastes, discuss the acidification problems and solutions in applying anaerobic digestion for fruit and vegetable wastes and investigate the reactor design (comparing single phase with two phase) and the thermal pre-treatment for processing raw wastes. Furthermore, it analyses the dominant microorganisms involved at different stages of digestion and suggests a focus for future studies.

Making lignin accessible for anaerobic digestion by wet-explosion pretreatment

DEFF Research Database (Denmark)

Ahring, Birgitte Kiær; Biswas, Rajib; Ahamed, Aftab

2015-01-01

of lignin during anaerobic digestion processes. The pretreatment of feedlot manure was performed in a 10 L reactor at 170 C for 25 min using 4 bars oxygen and the material was fed to a continuous stirred tank reactor operated at 55 C for anaerobic digestion. Methane yield of untreated and pretreated...... material was 70 ± 27 and 320 ± 36 L/kg-VS/day, respectively, or 4.5 times higher yield as a result of the pretreatment. Aliphatic acids formed during the pretreatment were utilized by microbes. 44.4% lignin in pretreated material was actually converted in the anaerobic digestion process compared to 12...

Recycling carbon dioxide during xylose fermentation by engineered Saccharomyces cerevisiae

Science.gov (United States)

In this study, we introduced the ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) and phosphoribulokinase (PRK) into an engineered S. cerevisiae (SR8) harboring the XR/XDH pathway and up-regulated PPP 10, to enable CO2 recycling through a synthetic rPPP during xylose fermentation (Fig. 1). ...

Electrochemistry for the Generation of Renewable Chemicals: One-Pot Electrochemical Deoxygenation of Xylose to δ-Valerolactone.

Science.gov (United States)

James, Olusola O; Sauter, Waldemer; Schröder, Uwe

2017-05-09

In this study, the electrochemical conversion of xylose to δ-valerolactone via carbonyl intermediates is demonstrated. The conversion was achieved in aqueous media and at ambient conditions. This study also demonstrates that the feedstock for production of renewable chemicals and biofuels through electrochemistry can be extended to primary carbohydrate molecules. This is the first report on a one-pot electrochemical deoxygenation of xylose to δ-valerolactone. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Co-fermentation of cellobiose and xylose by mixed culture of recombinant Saccharomyces cerevisiae and kinetic modeling.

Science.gov (United States)

Chen, Yingying; Wu, Ying; Zhu, Baotong; Zhang, Guanyu; Wei, Na

2018-01-01

Efficient conversion of cellulosic sugars in cellulosic hydrolysates is important for economically viable production of biofuels from lignocellulosic biomass, but the goal remains a critical challenge. The present study reports a new approach for simultaneous fermentation of cellobiose and xylose by using the co-culture consisting of recombinant Saccharomyces cerevisiae specialist strains. The co-culture system can provide competitive advantage of modularity compared to the single culture system and can be tuned to deal with fluctuations in feedstock composition to achieve robust and cost-effective biofuel production. This study characterized fermentation kinetics of the recombinant cellobiose-consuming S. cerevisiae strain EJ2, xylose-consuming S. cerevisiae strain SR8, and their co-culture. The motivation for kinetic modeling was to provide guidance and prediction of using the co-culture system for simultaneous fermentation of mixed sugars with adjustable biomass of each specialist strain under different substrate concentrations. The kinetic model for the co-culture system was developed based on the pure culture models and incorporated the effects of product inhibition, initial substrate concentration and inoculum size. The model simulations were validated by results from independent fermentation experiments under different substrate conditions, and good agreement was found between model predictions and experimental data from batch fermentation of cellobiose, xylose and their mixtures. Additionally, with the guidance of model prediction, simultaneous co-fermentation of 60 g/L cellobiose and 20 g/L xylose was achieved with the initial cell densities of 0.45 g dry cell weight /L for EJ2 and 0.9 g dry cell weight /L SR8. The results demonstrated that the kinetic modeling could be used to guide the design and optimization of yeast co-culture conditions for achieving simultaneous fermentation of cellobiose and xylose with improved ethanol productivity, which is

[Anaerobic bacteria isolated from patients with suspected anaerobic infections].

Science.gov (United States)

Ercis, Serpil; Tunçkanat, Ferda; Hasçelik, Gülşen

2005-10-01

The study involved 394 clinical samples sent to the Clinical Microbiology Laboratory of Hacettepe University Adult Hospital between January 1997 and May 2004 for anaerobic cultivation. Since multiple cultures from the same clinical samples of the same patient were excluded, the study was carried on 367 samples. The anaerobic cultures were performed in anaerobic jar using AnaeroGen kits (Oxoid, Basingstoke, U.K.) or GENbox (bioMérieux, Lyon, France). The isolates were identified by both classical methods and "BBL Crystal System" (Becton Dickinson, U.S.A.). While no growth was detected in 120 (32.7%) of the clinical samples studied, in 144 samples (39.2%) only aerobes, in 28 (7.6%) only anaerobes and in 75 (20.5%) of the samples both aerobes and anaerobes were isolated. The number of the anaerobic isolates was 217 from 103 samples with anaerobic growth. Of these 103 samples 15 showed single bacterial growth whereas in 88 samples multiple bacterial isolates were detected. Anaerobic isolates consisted of 92 Gram negative bacilli (Bacteroides spp. 50, Prevotella spp. 14, Porphyromonas spp. 10, Fusobacterium spp. 7, Tisierella spp. 2, unidentified 9), 57 Gram positive bacilli (Clostridium spp.17, Propionibacterium spp. 16, Lactobacillus spp. 8, Actinomyces spp. 5, Eubacterium spp. 2, Bifidobacterium adolescentis 1, Mobiluncus mulieris 1, unidentified nonspore forming rods 7), 61 Gram positive cocci (anaerobic cocci 44, microaerophilic cocci 17), and 7 Gram negative cocci (Veillonella spp.). In conclusion, in the samples studied with prediagnosis of anaerobic infection, Bacteroides spp. (23%) were the most common bacteria followed by anaerobic Gram positive cocci (20.3%) and Clostridium spp (7.8%).

In situ detection of anaerobic alkane metabolites in subsurface environments

Directory of Open Access Journals (Sweden)

Lisa eGieg

2013-06-01

Full Text Available Alkanes comprise a substantial fraction of crude oil and refined fuels. As such, they are prevalent within deep subsurface fossil fuel deposits and in shallow subsurface environments such as aquifers that are contaminated with hydrocarbons. These environments are typically anaerobic, and host diverse microbial communities that can potentially use alkanes as substrates. Anaerobic alkane biodegradation has been reported to occur under nitrate-reducing, sulfate-reducing, and methanogenic conditions. Elucidating the pathways of anaerobic alkane metabolism has been of interest in order to understand how microbes can be used to remediate contaminated sites. Alkane activation primarily occurs by addition to fumarate, yielding alkylsuccinates, unique anaerobic metabolites that can be used to indicate in situ anaerobic alkane metabolism. These metabolites have been detected in hydrocarbon-contaminated shallow aquifers, offering strong evidence for intrinsic anaerobic bioremediation. Recently, studies have also revealed that alkylsuccinates are present in oil and coal seam production waters, indicating that anaerobic microbial communities can utilize alkanes in these deeper subsurface environments. In many crude oil reservoirs, the in situ anaerobic metabolism of hydrocarbons such as alkanes may be contibuting to modern-day detrimental effects such as oilfield souring, or may lead to more benefical technologies such as enhanced energy recovery from mature oilfields. In this review, we briefly describe the key metabolic pathways for anaerobic alkane (including n-alkanes, isoalkanes, and cyclic alkanes metabolism and highlight several field reports wherein alkylsuccinates have provided evidence for anaerobic in situ alkane metabolism in shallow and deep subsurface environments.

Using contaminated plants involved in phytoremediation for anaerobic digestion.

Science.gov (United States)

Cao, Zewei; Wang, Shengxiao; Wang, Ting; Chang, Zhizhou; Shen, Zhenguo; Chen, Yahua

2015-01-01

This study investigated the anaerobic digestion capability of five plants and the effects of copper (Cu) and S,S'-ethylenediaminedisuccinic acid (EDDS, a chelator widely used in chelant-assisted phytoremediation) on biogas production to determine a feasible disposal method for plants used in remediation. The results showed that in addition to Phytolacca americana L., plants such as Zea mays L., Brassica napus L., Elsholtzia splendens Nakai ex F. Maekawa, and Oenothera biennis L. performed well in biogas production. Among these, O. biennis required the shortest period to finish anaerobic digestion. Compared to normal plants with low Cu content, the plants used in remediation with increased Cu levels (100 mg kg(-1)) not only promoted anaerobic digestion and required a shorter anaerobic digestion time, but also increased the methane content in biogas. When the Cu content in plants increased to 500, 1000, and 5000 mg kg(-1), the cumulative biogas production decreased by 12.3%, 14.6%, and 41.2%, respectively. Studies also found that EDDS conspicuously restrained biogas production from anaerobic digestion. The results suggest that anaerobic digestion has great potential for the disposal of contaminated plants and may provide a solution for the resource utilization of plants used in remediation.

Study on the Requirement of Nitrogen Sources by Scheffersomyces Stipitis NRRL Y-7124 to Produce Ethanol from Xylose Based-media

DEFF Research Database (Denmark)

Mussatto, Solange I.; Carneiro, L. M.; Roberto, I. C.

This study aimed at evaluating the requirement of nitrogen sources by the yeast Scheffersomyces stipitis NRRL Y-7124 to produce ethanol from xylose based-media. Different nitrogen sources were evaluated, which were used to supplement a defined xylose-based medium and also the hemicellulosic hydro...

Ethanol from Cellulosic Biomass with Emphasis of Wheat Straw Utilization. Analysis of Strategies for Process Development

Directory of Open Access Journals (Sweden)

Alexander Dimitrov Kroumov

2015-12-01

Full Text Available The "Green and Blue Technologies Strategies in HORIZON 2020" has increased the attention of scientific society on global utilization of renewable energy sources. Agricultural residues can be a valuable source of energy because of drastically growing human needs for food. The goal of this review is to show the current state of art on utilization of wheat straw as a substrate for ethanol production. The specifics of wheat straw composition and the chemical and thermodynamic properties of its components pre-determined the application of unit operations and engineering strategies for hydrolysis of the substrate and further its fermentation. Modeling of this two processes is crucially important for optimal overall process development and scale up. The authors gave much attention on main hydrolisis products as a glucose and xylose (C6 and C5 sugars, respectivelly and on the specifics of their metabolization by ethanol producing microorganisms. The microbial physiology reacting on C6 and C5 sugars and mathematical aproaches describing these phenomena are discussing, as well.

Adaptation of the xylose fermenting yeast Saccharomyces cerevisiae F12 for improving ethanol production in different fed-batch SSF processes.

Science.gov (United States)

Tomás-Pejó, E; Ballesteros, M; Oliva, J M; Olsson, L

2010-11-01

An efficient fermenting microorganism for bioethanol production from lignocellulose is highly tolerant to the inhibitors released during pretreatment and is able to ferment efficiently both glucose and xylose. In this study, directed evolution was employed to improve the xylose fermenting Saccharomyces cerevisiae F12 strain for bioethanol production at high substrate loading. Adapted and parental strains were compared with respect to xylose consumption and ethanol production. Adaptation led to an evolved strain more tolerant to the toxic compounds present in the medium. When using concentrated prehydrolysate from steam-pretreated wheat straw with high inhibitor concentration, an improvement of 65 and 20% in xylose consumption and final ethanol concentration, respectively, were achieved using the adapted strain. To address the need of high substrate loadings, fed-batch SSF experiments were performed and an ethanol concentration as high as 27.4 g/l (61% of the theoretical) was obtained with 11.25% (w/w) of water insoluble solids (WIS).

Implementation of an UASB anaerobic digester at bagasse-based pulp and paper industry

International Nuclear Information System (INIS)

Chinnaraj, S.; Venkoba Rao, G.

2006-01-01

Upflow anaerobic sludge blanket (UASB) reactor was installed to replace the conventional anaerobic lagoon treating bagasse wash wastewater from agro-based pulp and paper mill, to generate bio-energy and to reduce greenhouse gas emissions. The plant was designed to treat 12 ML d -1 of wastewater having two 5 ML capacity reactors, 5.75 kg COD m -3 d -1 organic loading rate and 20 h hydraulic retention time. In the plant 80-85% COD reduction was achieved with biogas production factor of 520 L kg -1 COD reduced. In 11 months 4.4 million m 3 of biogas was generated from bagasse wash wastewater utilizing UASB process. Utilization of the biogas in the Lime Kiln saved 2.14 ML of furnace oil in 9 months. Besides significant economic benefits, furnace oil saving reduced 6.4 Gg CO 2 emission from fossil fuel and conversion of the anaerobic lagoon into anaerobic reactor reduced 2.1 Gg methane emission which is equal to 43.8 Gg of CO 2

Microbial network for waste activated sludge cascade utilization in an integrated system of microbial electrolysis and anaerobic fermentation

DEFF Research Database (Denmark)

Liu, Wenzong; He, Zhangwei; Yang, Chunxue

2016-01-01

in an integrated system of microbial electrolysis cell (MEC) and anaerobic digestion (AD) for waste activated sludge (WAS). Microbial communities in integrated system would build a thorough energetic and metabolic interaction network regarding fermentation communities and electrode respiring communities...... to Firmicutes (Acetoanaerobium, Acetobacterium, and Fusibacter) showed synergistic relationship with exoelectrogensin the degradation of complex organic matter or recycling of MEC products (H2). High protein and polysaccharide but low fatty acid content led to the dominance of Proteiniclasticum...... biofilm. The overall performance of WAS cascade utilization was substantially related to the microbial community structures, which in turn depended on the initial pretreatment to enhance WAS fermentation. It is worth noting that species in AD and MEC communities are able to build complex networks...

Effects of sludge inoculum and organic feedstock on active microbial communities and methane yield during anaerobic digestion

Directory of Open Access Journals (Sweden)

David eWilkins

2015-10-01

Full Text Available Anaerobic digestion (AD is a widespread microbial technology used to treat organic waste and recover energy in the form of methane (biogas. While most AD systems have been designed to treat a single input, mixtures of digester sludge and solid organic waste are emerging as a means to improve efficiency and methane yield. We examined laboratory anaerobic cultures of AD sludge from two sources amended with food waste, xylose, and xylan at mesophilic temperatures, and with cellulose at meso- and thermophilic temperatures, to determine whether and how the inoculum and substrate affect biogas yield and community composition. All substrate and inoculum combinations yielded methane, with food waste most productive by mass. Pyrosequencing of transcribed bacterial and archaeal 16S rRNA showed that community composition varied across substrates and inocula, with differing ratios of hydrogenotrophic/acetoclastic methanogenic archaea associated with syntrophic partners. While communities did not cluster by either inoculum or substrate, additional sequencing of the bacterial 16S rRNA gene in the source sludge revealed that the bacterial communities were influenced by their inoculum. These results suggest that complete and efficient AD systems could potentially be assembled from different microbial inocula and consist of taxonomically diverse communities that nevertheless perform similar functions.

Anaerobic bacterium that degrades fatty acids in syntrophic association with methanogens

Energy Technology Data Exchange (ETDEWEB)

McInerney, M J [Univ. of Illinois, Urbana; Bryant, M P; Pfennig, N

1979-01-01

A new species of anaerobic bacterium that degrades the even-numbered carbon fatty acids, butyrate, caproate and caprylate, to acetate and H/sub 2/ and the odd-numbered carbon fatty acids, valerate and heptanoate, to acetate, propionate and H/sub 2/ was obtained in coculture with either an H/sub 2/-utilizing methanogen or H/sub 2/-utilizing desulfovibrio. The organism could be grown only in syntrophic association with the H/sub 2/-utilizer and no other energy sources or combination of electron donor and acceptors were utilized. It was a Gram-negative helical rod with 2 to 8 flagella, about 20 nm in diameter, inserted in a linear fashion about 130 nm or more apart along the concave side of the cell. It grew with a generation time of 84 h in co-culture with Methanospirillum hungatii and was present in numbers of at least 4.5 x 10/sup -6/ per g of anaerobic digest or sludge.

Timeline of bio-hydrogen production by anaerobic digestion of biomass

Directory of Open Access Journals (Sweden)

Bernadette E. TELEKY

2015-12-01

Full Text Available Anaerobic digestion of biomass is a process capable to produce biohydrogen, a clean source of alternative energy. Lignocellulosic biomass from agricultural waste is considered a renewable energy source; therefore its utilization also contributes to the reduction of water, soil and air pollution. The study consists in five consecutive experiments designed to utilize anaerobic bacterial enrichment cultures originating from the Hungarian Lake, Hévíz. Wheat straw was used as complex substrate to produce hydrogen. The timeline evolution of hydrogen production was analyzed and modelled by two functions: Logistic and Boltzmann. The results proved that hydrogen production is significant, with a maximum of 0.24 mlN/ml and the highest hydrogen production occurs between the days 4-10 of the experiment.

Improvement of ACE inhibitory activity of casein hydrolysate by Maillard reaction with xylose.

Science.gov (United States)

Hong, Xu; Meng, Jun; Lu, Rong-Rong

2015-01-01

The Maillard reaction is widely used to improve the functional properties or biological activities of food. The purpose of this study was to investigate the effect of the Maillard reaction on angiotensin I converting enzyme (ACE) inhibitory activity in a casein hydrolysate-xylose system. Two-step hydrolysis was used to prepare casein ACE inhibitory peptides. Maillard reaction products (MRPs) were prepared by heating hydrolyzed casein with xylose at pH 8.0, 110 °C for up to 16 h. The results showed that the content of free amino group decreased (P Maillard reaction (P reaction in the MRPs. The study shows that the Maillard reaction under appropriate conditions can improve the ACE inhibitory activity of casein hydrolysate effectively. © 2014 Society of Chemical Industry.

Dehydration of D-xylose to furfural using acid-functionalized MWCNTs catalysts

Science.gov (United States)

Termvidchakorn, Chompoopitch; Itthibenchapong, Vorranutch; Songtawee, Siripit; Chamnankid, Busaya; Namuangruk, Supawadee; Faungnawakij, Kajornsak; Charinpanitkul, Tawatchai; Khunchit, Radchadaporn; Hansupaluk, Nanthiya; Sano, Noriaki; Hinode, Hirofumi

2017-09-01

Acid-functionalized multi-wall carbon nanotubes (MWCNTs) catalysts were prepared by a wet chemical sonication with various acid solutions, i.e. H2SO4, H3PO4, HNO3, and HCl. Sulfonic groups and carboxyl groups were detected on MWCNTs with H2SO4 treatment (s-MWCNTs), while only carboxyl groups were presented from other acid treatments. The catalytic dehydration of D-xylose into furfural was evaluated using a batch reactor at 170 °C for 3 h under N2 pressure of 15 bar. The highest furfural selectivity was achieved around 57% by s-MWCNTs catalyst, suggesting a positive role of the sulfonic functionalized groups. The effect of Co species was related to their Lewis acid property resulting in the enhancement of xylose conversion with low selectivity to furfural product. Invited talk at 5th Thailand International Nanotechnology Conference (Nano Thailand-2016), 27-29 November 2016, Nakhon Ratchasima, Thailand.

Dehydration of D-xylose over SiO2-Al2O3 catalyst: Perspective on the pathways for condensed products

International Nuclear Information System (INIS)

You, Su Jin; Park, Eun Duck; Park, Myung-June

2016-01-01

This work addresses the kinetic mechanism for the dehydration of D-xylose over the SiO 2 -Al 2 O 3 solid catalyst, where the formation of condensed products is included in addition to the production of furfural and its decomposition. The kinetic modeling and parametric sensitivity show that the isomerization of D-xylose takes place in the early stages of the reaction, followed by the dehydration of isomers. Accordingly, the homogeneous polymerization of isomers is found to be dominant. The developed model is used to evaluate the effects of operating conditions on the catalytic performance; high temperature and D-xylose concentration guarantee high furfural yield.

Gelria glutamica gen. nov., sp. a thermophilic oligately syntrophic glutamate-degrading anaerobe

NARCIS (Netherlands)

Plugge, C.M.; Balk, M.; Zoetendal, E.G.; Stams, A.J.M.

2002-01-01

A novel anaerobic, Gram-positive, thermophilic, spore-forming, obligately syntrophic, glutamate-degrading bacterium, strain TGO(T), was isolated from a propionate-oxidizing methanogenic enrichment culture. The axenic culture was obtained by growing the bacterium on pyruvate. Cells were rod-shaped

DEVELOPMENT OF IMPROVED ANAEROBIC GROWTH OF BACILLUS MOJAVENSIS STRAIN JF-2 FOR THE PURPOSE OF IMPROVED ANAEROBIC BIOSURFACTANT PRODUCTION FOR ENHANCED OIL RECOVERY

Energy Technology Data Exchange (ETDEWEB)

M.J. McInerney; M. Folmsbee; D. Nagle

2004-05-31

Our work focuses on the use of microorganisms to recover petroleum hydrocarbons that remain entrapped after current recovery technologies reach their economic limit. Capillary forces between the hydrocarbon and aqueous phases are largely responsible for trapping the hydrocarbons in the pores of the rock and large reductions in the interfacial tension between the hydrocarbon and aqueous phases are needed for hydrocarbon mobilization (1-3, 10, 11). Microorganisms produce a variety of biosurfactants (4), several of which generate the ultra low interfacial tensions needed for hydrocarbon mobilization (4, 5, 8). In particular, the lipopeptide biosurfactant produced by Bacillus mojavensis strain JF-2 reduces the interfacial tension between hydrocarbon and aqueous phases to very low levels (<0.016 mN/m) (8) (9). B. mojavensis JF-2 grows under the environmental conditions found in many oil reservoirs, i. e., anaerobic, NaCl concentrations up to 80 g l{sup -1}, and temperatures up to 45 C (6, 7), making it ideally suited for in situ applications. However, anaerobic growth of B. mojavensis JF-2 was inconsistent and difficult to replicate, which limited its use for in situ applications. Our initial studies revealed that enzymatic digests, such as Proteose Peptone, were required for anaerobic growth of Bacillus mojavensis JF-2. Subsequent purification of the growth-enhancing factor in Proteose Peptone resulted in the identification of the growth-enhancing factor as DNA or deoxyribonucleosides. The addition of salmon sperm DNA, herring sperm DNA, E. coli DNA or synthetic DNA (single or double stranded) to Medium E all supported anaerobic growth of JF-2. Further, we found that JF-2 required all four deoxyribonucleosides (deoxyadeonosine, deoxyguanosine, deoxycytidine and thymidine) for growth under strict anaerobic conditions. The requirement for the deoxyribonucleosides did not occur under aerobic growth conditions. DNA was not used as a sole energy source; sucrose was required

Potential for using thermophilic anaerobic bacteria for bioethanol production from hemicellulose

DEFF Research Database (Denmark)

Sommer, P.; Georgieva, Tania I.; Ahring, Birgitte Kiær

2004-01-01

A limited number of bacteria, yeast and fungi can convert hemicellulose or its monomers (xylose, arabinose, mannose and galactose) into ethanol with a satisfactory yield and productivity. In the present study we tested a number of thermophilic enrichment cultures, and new isolates of thermophilic...... Of D-Xylose into ethanol; (ii) test for viability and ethanol production in pretreated wheat straw hemicellulose hydrolysate; (iii) test for tolerance against high D-xylose concentrations. A total of 86 enrichment cultures and 58 pure cultures were tested and five candidates were selected which...

Growing Chlorella vulgaris on thermophilic anaerobic digestion swine manure for nutrient removal and biomass production.

Science.gov (United States)

Deng, Xiang-Yuan; Gao, Kun; Zhang, Ren-Chuan; Addy, Min; Lu, Qian; Ren, Hong-Yan; Chen, Paul; Liu, Yu-Huan; Ruan, Roger

2017-11-01

Liquid swine manure was subjected to thermophilic anaerobic digestion, ammonia stripping and centrifugation in order to increase the available carbon sources and decrease the ammonia concentration and turbidity. Chlorella vulgaris (UTEX 2714) was grown on minimally diluted (2×, 3× and 4×) autoclaved and non-autoclaved pretreated anaerobic digestion swine manure (PADSM) in a batch-culture system for 7days. Results showed that C. vulgaris (UTEX 2714) grew best on 3× PADSM media, and effectively removed NH 4 + -N, TN, TP and COD by 98.5-99.8%, 49.2-55.4%, 20.0-29.7%, 31.2-34.0% and 99.8-99.9%, 67.4-70.8%, 49.3-54.4%, 73.6-78.7% in differently diluted autoclaved and non-autoclaved PADSM, respectively. Results of chemical compositions indicated that contents of pigment, carbohydrate, protein and lipid in C. vulgaris (UTEX 2714) changed with the culture conditions. Moreover, its fatty acid profiles suggested that this alga could be used as animal feed if cultivated in autoclaved PADSM or as good-quality biodiesel feedstock if cultivated in non-autoclaved PADSM. Copyright © 2017 Elsevier Ltd. All rights reserved.

How to isolate, identify and determine antimicrobial susceptibility of anaerobic bacteria in routine laboratories?

Science.gov (United States)

Nagy, E; Boyanova, L; Justesen, U S

2018-02-17

There has been increased interest in the study of anaerobic bacteria that cause human infection during the past decade. Many new genera and species have been described using 16S rRNA gene sequencing of clinical isolates obtained from different infection sites with commercially available special culture media to support the growth of anaerobes. Several systems, such as anaerobic pouches, boxes, jars and chambers provide suitable anaerobic culture conditions to isolate even strict anaerobic bacteria successfully from clinical specimens. Beside the classical, time-consuming identification methods and automated biochemical tests, the use of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry has revolutionized identification of even unusual and slow-growing anaerobes directly from culture plates, providing the possibility of providing timely information about anaerobic infections. The aim of this review article is to present methods for routine laboratories, which carry out anaerobic diagnostics on different levels. Relevant data from the literature mostly published during the last 7 years are encompassed and discussed. The review involves topics on the anaerobes that are members of the commensal microbiota and their role causing infection, the key requirements for collection and transport of specimens, processing of specimens in the laboratory, incubation techniques, identification and antimicrobial susceptibility testing of anaerobic bacteria. Advantages, drawbacks and specific benefits of the methods are highlighted. The present review aims to update and improve anaerobic microbiology in laboratories with optimal conditions as well as encourage its routine implementation in laboratories with restricted resources. Copyright © 2018 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

Towards a Microbial Production of Fatty Acids as Precursors of Biokerosene from Glucose and Xylose Vers une production microbienne d’acides gras en vue de l’application biokérosène à partir de glucose et xylose

Directory of Open Access Journals (Sweden)

Babau M.

2013-09-01

Full Text Available The aviation industry considers the development of sustainable biofuels as one of the biggest challenges of the next ten years. The aim is to lower the environmental impact of the steadily increasing use of fossil fuels on climate change, yielding greater energy independence and fuel security. Thus, the development of a new route for the production of lipids from renewable non-food resources is now being promoted with the recent ASTM certification of hydrotreated oils. Our study focuses on the potential of growth of the oleaginous yeast Rhodotorula glutinis using glucose and xylose which can come from renewable lignocellulosic substrates and of lipid accumulation using glucose as substrate. Experiments were carried out in fed-batch mode which allowed feed flux management. Carbon fluxes were controlled with modifying xylose/glucose ratios to quantify metabolism in optimal growth condition. Besides, the management of carbon and nitrogen fluxes allowed characterizing lipid accumulation. Thus, it has been shown that the yeast Rhodotorula glutinis can simultaneously consume glucose and xylose. When the ratio xylose/glucose increased, the growth rate and the carbon conversion yield into biomass decreased: it was of 0.36 h-1 and 0.64 Cmol x*.Cmol glu-1 for pure glucose, it was of 0.15 h-1 and 0.56 Cmol.Cmol-1 for 10% xylose and it was of 0.037 h-1 and 0.18 Cmol.Cmol-1 for pure xylose. The necessity to maintain residual growth and to manage carbon fluxes to optimize lipid accumulation performance was revealed. Lipid accumulation on glucose engendered a final biomass concentration of 150 gCDW.L-1, microbial production (72% of lipids and maximal productivity over 1.48 glip.L-1.h-1. The culture temperature is an important parameter to modulate the lipid profile. The results were encouraging. Lipid accumulation using lignocellulosic feedstock was shown to be a highly promising route. Le développement de filières de production de molécules �

Microbial production of bulk chemicals: development of anaerobic processes

NARCIS (Netherlands)

Weusthuis, R.A.; Lamot, I.; Oost, van der J.; Sanders, J.P.M.

2011-01-01

nnovative fermentation processes are necessary for the cost-effective production of bulk chemicals from renewable resources. Current microbial processes are either anaerobic processes, with high yield and productivity, or less-efficient aerobic processes. Oxygen utilization plays an important role

Methane-yielding microbial communities processing lactate-rich substrates: a piece of the anaerobic digestion puzzle.

Science.gov (United States)

Detman, Anna; Mielecki, Damian; Pleśniak, Łukasz; Bucha, Michał; Janiga, Marek; Matyasik, Irena; Chojnacka, Aleksandra; Jędrysek, Mariusz-Orion; Błaszczyk, Mieczysław K; Sikora, Anna

2018-01-01

Anaerobic digestion, whose final products are methane and carbon dioxide, ensures energy flow and circulation of matter in ecosystems. This naturally occurring process is used for the production of renewable energy from biomass. Lactate, a common product of acidic fermentation, is a key intermediate in anaerobic digestion of biomass in the environment and biogas plants. Effective utilization of lactate has been observed in many experimental approaches used to study anaerobic digestion. Interestingly, anaerobic lactate oxidation and lactate oxidizers as a physiological group in methane-yielding microbial communities have not received enough attention in the context of the acetogenic step of anaerobic digestion. This study focuses on metabolic transformation of lactate during the acetogenic and methanogenic steps of anaerobic digestion in methane-yielding bioreactors. Methane-yielding microbial communities instead of pure cultures of acetate producers were used to process artificial lactate-rich media to methane and carbon dioxide in up-flow anaerobic sludge blanket reactors. The media imitated the mixture of acidic products found in anaerobic environments/digesters where lactate fermentation dominates in acidogenesis. Effective utilization of lactate and biogas production was observed. 16S rRNA profiling was used to examine the selected methane-yielding communities. Among Archaea present in the bioreactors, the order Methanosarcinales predominated. The acetoclastic pathway of methane formation was further confirmed by analysis of the stable carbon isotope composition of methane and carbon dioxide. The domain Bacteria was represented by Bacteroidetes , Firmicutes , Proteobacteria , Synergistetes , Actinobacteria , Spirochaetes , Tenericutes , Caldithrix , Verrucomicrobia , Thermotogae , Chloroflexi , Nitrospirae, and Cyanobacteria. Available genome sequences of species and/or genera identified in the microbial communities were searched for genes encoding the lactate

A formal synthesis of (+-muricatacin from D-xylose

Directory of Open Access Journals (Sweden)

VELIMIR POPSAVIN

2003-11-01

Full Text Available A multistep route towards the aldehydo-lactone 19, the final chiral precursor in a new stereospecific synthesis of (+-muricatacin, has been developed starting from D-xylose. The key step of the synthesis involves an E-selective Wittig olefination of the lactol 6 with methoxycarbonylmethylidene triphenylphosphorane, followed by successive catalytic reduction and g-lactonisation processes. Subsequent selective functional groups interconversions furnished the key six-carbon intermediate 19, which can be converted into the (+-muricatacin via a three-step sequence already described in the chemical literature.

The future of anaerobic digestion and biogas utilization

DEFF Research Database (Denmark)

Holm-Nielsen, J.B.; Al Seadi, T.; Oleskowicz-Popiel, Piotr

2009-01-01

One of the common tendencies of animal production activities in Europe and in developed countries in general is to intensify the animal production and to increase the size of the animal production units. High livestock density is always accompanied by production of a surplus of animal manure...... and to redistribute the excess of nutrients from manure and to optimize their recycling. Anaerobic digestion of animal manure and slurries offers several benefits by improving their fertilizer qualities, reducing odors and pathogens and producing a renewable fuel – the biogas. The EU policies concerning renewable...... energy systems (RES) have set forward a fixed goal of supplying 20% of the European energy demands from RES by year 2020. A major part of the renewable energy will originate from European farming and forestry. At least 25% of all bioenergy in the future can originate from biogas, produced from wet...

Automated UV-C mutagenesis of Kluyveromyces marxianus NRRL Y-1109 and selection for microaerophilic growth and ethanol production at elevated temperature on biomass sugars.

Science.gov (United States)

Hughes, Stephen R; Bang, Sookie S; Cox, Elby J; Schoepke, Andrew; Ochwat, Kate; Pinkelman, Rebecca; Nelson, Danielle; Qureshi, Nasib; Gibbons, William R; Kurtzman, Cletus P; Bischoff, Kenneth M; Liu, Siqing; Cote, Gregory L; Rich, Joseph O; Jones, Marjorie A; Cedeño, David; Doran-Peterson, Joy; Riaño-Herrera, Nestor M; Rodríguez-Valencia, Nelson; López-Núñez, Juan C

2013-08-01

The yeast Kluyveromyces marxianus is a potential microbial catalyst for fuel ethanol production from a wide range of biomass substrates. To improve its growth and ethanol yield at elevated temperature under microaerophilic conditions, K. marxianus NRRL Y-1109 was irradiated with UV-C using automated protocols on a robotic platform for picking and spreading irradiated cultures and for processing the resulting plates. The plates were incubated under anaerobic conditions on xylose or glucose for 5 mo at 46 °C. Two K. marxianus mutant strains (designated 7-1 and 8-1) survived and were isolated from the glucose plates. Both mutant strains, but not wild type, grew aerobically on glucose at 47 °C. All strains grew anaerobically at 46 °C on glucose, galactose, galacturonic acid, and pectin; however, only 7-1 grew anaerobically on xylose at 46 °C. Saccharomyces cerevisiae NRRL Y-2403 did not grow at 46 °C on any of these substrates. With glucose as a carbon source, ethanol yield after 3 d at 46 °C was higher for 8-1 than for wild type (0.51 and 0.43 g ethanol/g glucose, respectively). With galacturonic acid as a carbon source, the ethanol yield after 7 d at 46 °C was higher for 7-1 than for wild type (0.48 and 0.34 g ethanol/g galacturonic acid, respectively). These mutant strains have potential application in fuel ethanol production at elevated temperature from sugar constituents of starch, sucrose, pectin, and cellulosic biomass.

Catalytic conversion of xylose and corn stalk into furfural over carbon solid acid catalyst in γ-valerolactone.

Science.gov (United States)

Zhang, Tingwei; Li, Wenzhi; Xu, Zhiping; Liu, Qiyu; Ma, Qiaozhi; Jameel, Hasan; Chang, Hou-min; Ma, Longlong

2016-06-01

A novel carbon solid acid catalyst was synthesized by the sulfonation of carbonaceous material which was prepared by carbonization of sucrose using 4-BDS as a sulfonating agent. TEM, N2 adsorption-desorption, elemental analysis, XPS and FT-IR were used to characterize the catalyst. Then, the catalyst was applied for the conversion of xylose and corn stalk into furfural in GVL. The influence of the reaction time, temperature and dosage of catalyst on xylose dehydration were also investigated. The Brønsted acid catalyst exhibited high activity in the dehydration of xylose, with a high furfural yield of 78.5% at 170°C in 30min. What's more, a 60.6% furfural yield from corn stalk was achieved in 100min at 200°C. The recyclability of the sulfonated carbon catalyst was perfect, and it could be reused for 5times without the loss of furfural yields. Copyright © 2016 Elsevier Ltd. All rights reserved.

Anaerobic digestion and related best management practices : utilizing life cycle assessment

Energy Technology Data Exchange (ETDEWEB)

Venczel, M.Z. [Clarkson Univ., Potsdam, NY (United States); Powers, S.E. [Clarkson Univ., Potsdam, NY (United States)

2010-07-01

This paper reported on a life cycle assessment (LCA) study that compared the environmental impacts of business-as-usual manure management with those of a manure management operation incorporating anaerobic digestion with combined heat and power generation. The case study was based on a medium sized dairy farm in northern New York State. The study identified the benefits resulting from the displacement of fossil fuels, and reduction of related emissions. Although anaerobic digestion of dairy manure with energy recovery through biogas combustion is viewed as a positive environmental approach to increase the use of renewable energy, there are potential negative impacts that can counteract the environmental benefits. The negative impacts are associated with emissions of methane and nitrogen species during digestion and after spreading of digester effluent. The environmental impacts and their causes should be evaluated in order to promote best management practices. Knowledge gained from an LCA was used in this study to assess the benefits associated with various management practices. The study showed that the design and construction of biogas systems must minimize the potential for fugitive emissions of biogas that can readily outweigh the benefits associated fossil fuel displacement. The environmental trade-offs associated with various manure management and energy recovery systems were also described.

Highly efficient production of L-lactic acid from xylose by newly isolated Bacillus coagulans C106.

Science.gov (United States)

Ye, Lidan; Zhou, Xingding; Hudari, Mohammad Sufian Bin; Li, Zhi; Wu, Jin Chuan

2013-03-01

Cost-effective production of optically pure lactic acid from lignocellulose sugars is commercially attractive but challenging. Bacillus coagulans C106 was isolated from environment and used to produce l-lactic acid from xylose at 50°C and pH 6.0 in mineral salts medium containing 1-2% (w/v) of yeast extract without sterilizing the medium before fermentation. In batch fermentation with 85g/L of xylose, lactic acid titer and productivity reached 83.6g/L and 7.5g/Lh, respectively. When fed-batch (120+80+60g/L) fermentation was applied, they reached 215.7g/L and 4.0g/Lh, respectively. In both cases, the lactic acid yield and optical purity reached 95% and 99.6%, respectively. The lactic acid titer and productivity on xylose are the highest among those ever reported. Ca(OH)2 was found to be a better neutralizing agent than NaOH in terms of its giving higher lactic acid titer (1.2-fold) and productivity (1.8-fold) under the same conditions. Copyright © 2013 Elsevier Ltd. All rights reserved.

Anaerobic Digestion: Process

DEFF Research Database (Denmark)

Angelidaki, Irini; Batstone, Damien J.

2011-01-01

Organic waste may degrade anaerobically in nature as well as in engineered systems. The latter is called anaerobic digestion or biogasification. Anaerobic digestion produces two main outputs: An energy-rich gas called biogas and an effluent. The effluent, which may be a solid as well as liquid...... with very little dry matter may also be called a digest. The digest should not be termed compost unless it specifically has been composted in an aerated step. This chapter describes the basic processes of anaerobic digestion. Chapter 9.5 describes the anaerobic treatment technologies, and Chapter 9...

Pathways and bioenergetics of anaerobic carbon monoxide fermentation

NARCIS (Netherlands)

Diender, Martijn; Stams, Fons; Machado de Sousa, Diana

2015-01-01

Carbon monoxide can act as a substrate for different modes of fermentative anaerobic metabolism. The trait of utilizing CO is spread among a diverse group of microorganisms, including members of bacteria as well as archaea. Over the last decade this metabolism has gained interest due to the

The kinetics of Scenedesmus obliquus microalgae growth utilizing carbon dioxide gas from biogas

International Nuclear Information System (INIS)

Thiansathit, Worrarat; Keener, Tim C.; Khang, Soon-Jai; Ratpukdi, Thunyalux; Hovichitr, Patcharee

2015-01-01

Microalgae Scenedesmus obliquus was cultured in a laboratory photobioreactor to determine the efficacy of using biogas as a carbon source for the microalgae's growth. The biogas contained ∼60% CH 4 and ∼40% CO 2 , and was derived from an anaerobic digester operating from animal wastes, and an anaerobic reactor utilizing high strength wastewater. The results showed that biogas is a viable carbon source for microalgae growth and that significant portions of the biogas' CO 2 can be utilized for algae growth, resulting in a biogas having a high concentration of methane. This paper develops the kinetic expressions for the algae's growth by assuming an autocatalytic reaction between carbon substrate and microalgae. The maximum specific growth rate and biomass productivity of S. obliquus were 0.56 d −1 and 0.145 g L −1 d −1 respectively. The biomass contained 51.8% carbon and higher heating value (HHV) was 22.9 MJ kg −1 . - Highlights: • Biogas is a viable carbon source for microalgae growth. • Biomass production rate and characteristics were assessed. • Scenedesmus obliquus can adjust to grow with high concentration of CO 2 in the carbon source

PENGOLAHAN LIMBAH CAIR INDUSTRI FARMASI FORMULASI DENGAN METODE ANAEROB-AEROB DAN ANAEROB-KOAGULASI

OpenAIRE

Farida Crisnaningtyas; Hanny Vistanty

2016-01-01

Studi ini membahas mengenai pengolahan limbah cair industri farmasi dalam skala laboratorium dengan menggunakan konsep anaerob-kimia-fisika dan anaerob-aerob. Proses anaerob dilakukan dengan menggunakan reaktor Upflow Anaerobic Sludge Bed reactor (UASBr) pada kisaran OLR (Organic Loading Rate) 0,5 – 2 kg COD/m3hari, yang didahului dengan proses aklimatisasi menggunakan substrat gula. Proses anaerob mampu memberikan efisiensi penurunan COD hingga 74%. Keluaran dari proses anaerob diolah lebih ...

Anaerobes in pleuropulmonary infections

Directory of Open Access Journals (Sweden)

De A

2002-01-01

Full Text Available A total of 76 anaerobes and 122 aerobes were isolated from 100 patients with pleuropulmonary infections, e.g. empyema (64, pleural effusion (19 and lung abscess (13. In 14% of the patients, only anaerobes were recovered, while a mixture of aerobes and anaerobes was encountered in 58%. From all cases of lung abscess, anaerobic bacteria were isolated, alone (04 or along with aerobic bacteria (13. From empyema and pleural effusion cases, 65.6% and 68.4% anaerobes were recovered respectively. Amongst anaerobes, gram negative anaerobic bacilli predominated (Prevotella melaninogenicus 16, Fusobacterium spp. 10, Bacteroides spp. 9, followed by gram positive anaerobic cocci (Peptostreptococcus spp. 31. Coliform bacteria (45 and Pseudomonas aeruginosa (42 were the predominant aerobic isolates.

The level of glucose-6-phosphate dehydrogenase activity strongly influences xylose fermentation and inhibitor sensitivity in recombinant Saccharomyces cerevisiae strains

DEFF Research Database (Denmark)

Jeppsson, M.; Johansson, B.; Jensen, Peter Ruhdal

2003-01-01

production levels of G6PDH on xylose fermentation. We used a synthetic promoter library and the copper-regulated CUP1 promoter to generate G6PDH-activities between 0% and 179% of the wildtype level. G6PDH-activities of 1% and 6% of the wild-type level resulted in 2.8- and 5.1-fold increase in specific xylose...

Optimised formation of blue Maillard reaction products of xylose and glycine model systems and associated antioxidant activity.

Science.gov (United States)

Yin, Zi; Sun, Qian; Zhang, Xi; Jing, Hao

2014-05-01

A blue colour can be formed in the xylose (Xyl) and glycine (Gly) Maillard reaction (MR) model system. However, there are fewer studies on the reaction conditions for the blue Maillard reaction products (MRPs). The objective of this study is to investigate characteristic colour formation and antioxidant activities in four different MR model systems and to determine the optimum reaction conditions for the blue colour formation in a Xyl-Gly MR model system, using the random centroid optimisation program. The blue colour with an absorbance peak at 630 nm appeared before browning in the Xyl-Gly MR model system, while no blue colour formation but only browning was observed in the xylose-alanine, xylose-aspartic acid and glucose-glycine MR model systems. The Xyl-Gly MR model system also showed higher antioxidant activity than the other three model systems. The optimum conditions for blue colour formation were as follows: xylose and glycine ratio 1:0.16 (M:M), 0.20 mol L⁻¹ NaHCO₃, 406.1 mL L⁻¹ ethanol, initial pH 8.63, 33.7°C for 22.06 h, which gave a much brighter blue colour and a higher peak at 630 nm. A characteristic blue colour could be formed in the Xyl-Gly MR model system and the optimum conditions for the blue colour formation were proposed and confirmed. © 2013 Society of Chemical Industry.

Automatic segmentation of MRI head images by 3-D region growing method which utilizes edge information

International Nuclear Information System (INIS)

Jiang, Hao; Suzuki, Hidetomo; Toriwaki, Jun-ichiro

1991-01-01

This paper presents a 3-D segmentation method that automatically extracts soft tissue from multi-sliced MRI head images. MRI produces a sequence of two-dimensional (2-D) images which contains three-dimensional (3-D) information of organs. To utilize such information we need effective algorithms to treat 3-D digital images and to extract organs and tissues of interest. We developed a method to extract the brain from MRI images which uses a region growing procedure and integrates information of uniformity of gray levels and information of the presence of edge segments in the local area around the pixel of interest. First we generate a kernel region which is a part of brain tissue by simple thresholding. Then we grow the region by means of a region growing algorithm under the control of 3-D edge existence to obtain the region of the brain. Our method is rather simple because it uses basic 3-D image processing techniques like spatial difference. It is robust for variation of gray levels inside a tissue since it also refers to the edge information in the process of region growing. Therefore, the method is flexible enough to be applicable to the segmentation of other images including soft tissues which have complicated shapes and fluctuation in gray levels. (author)

NADPH-dependent D-aldose reductases and xylose fermentation in Fusarium oxysporum

DEFF Research Database (Denmark)

Panagiotou, Gianni; Christakopoulos, P.

2004-01-01

Two aldose (xylose) reductases (ARI and ARII) from Fusarium oxysporum were purified and characterized. The native ARI was a monomer with M-r 41000, pI 5.2 and showed a 52-fold preference for NADPH over NADH, while ARII was homodimeric with a subunit of M-r 37000, pI 3.6 and a 60-fold preference...

Xylose-rich polysaccharides from the primary walls of embryogenic cell line of Pinus caribaea.

Science.gov (United States)

Mollard, A; Domon, J M; David, H; Joseleau, J P

1997-08-01

Embryogenic cell lines of Pinus caribaea were isolated from somatic embryogenesis from zygotic embryos. Previous studies showed that the proteins and glycoproteins were characteristic of the embryogenic state. In the present work we were seeking typical feature in the polysaccharide from the cell walls of embryogenic calli at nine days of culture. Sequential extraction with water, ammonium oxalate, dimethyl sulfoxide, sodium borohydride and 4.3 M potassium hydroxide revealed that the extracted polysaccharides contained high proportions of arabinose and significant amounts of xylose. Fractionation of the hydrosoluble polymers on DEAE cellulose afforded a xylose-rich fraction (80% xylose, 24% glucose and lower properties of fucose and mannose). Methylation analysis and 13C-NMR spectra showed that the glycan backbone consisted of beta 1 --> 4 linked xylosyl residues Similar study of the fractions extracted respectively with DMSO and 4.3 M KOH showed the presence of polydisperse glycoxylans but excluded the presence of xyloglucan in significant amount. This could be a characteristic feature of embryogenic cells walls of Pinus caribaea or could be typical of cells grown as calluses. In the various fractions obtained from DEAE cellulose chromatography of the alkaline extract the infrequent occurrence of fucoxylans beside an arabinogalactan showed again the unusual nature of the cell wall polymers of this embryogenic lines, which seems to differ greatly from those found in the primary wall of cells from suspension cultures.

Energy positive domestic wastewater treatment: the roles of anaerobic and phototrophic technologies.

Science.gov (United States)

Shoener, B D; Bradley, I M; Cusick, R D; Guest, J S

2014-05-01

The negative energy balance of wastewater treatment could be reversed if anaerobic technologies were implemented for organic carbon oxidation and phototrophic technologies were utilized for nutrient recovery. To characterize the potential for energy positive wastewater treatment by anaerobic and phototrophic biotechnologies we performed a comprehensive literature review and analysis, focusing on energy production (as kJ per capita per day and as kJ m(-3) of wastewater treated), energy consumption, and treatment efficacy. Anaerobic technologies included in this review were the anaerobic baffled reactor (ABR), anaerobic membrane bioreactor (AnMBR), anaerobic fluidized bed reactor (AFB), upflow anaerobic sludge blanket (UASB), anaerobic sequencing batch reactor (ASBR), microbial electrolysis cell (MEC), and microbial fuel cell (MFC). Phototrophic technologies included were the high rate algal pond (HRAP), photobioreactor (PBR), stirred tank reactor, waste stabilization pond (WSP), and algal turf scrubber (ATS). Average energy recovery efficiencies for anaerobic technologies ranged from 1.6% (MFC) to 47.5% (ABR). When including typical percent chemical oxygen demand (COD) removals by each technology, this range would equate to roughly 40-1200 kJ per capita per day or 110-3300 kJ m(-3) of treated wastewater. The average bioenergy feedstock production by phototrophic technologies ranged from 1200-4700 kJ per capita per day or 3400-13 000 kJ m(-3) (exceeding anaerobic technologies and, at times, the energetic content of the influent organic carbon), with usable energy production dependent upon downstream conversion to fuels. Energy consumption analysis showed that energy positive anaerobic wastewater treatment by emerging technologies would require significant reductions of parasitic losses from mechanical mixing and gas sparging. Technology targets and critical barriers for energy-producing technologies are identified, and the role of integrated anaerobic and

Energy positive domestic wastewater treatment: the roles of anaerobic and phototrophic technologies

KAUST Repository

Shoener, B. D.

2014-01-01

The negative energy balance of wastewater treatment could be reversed if anaerobic technologies were implemented for organic carbon oxidation and phototrophic technologies were utilized for nutrient recovery. To characterize the potential for energy positive wastewater treatment by anaerobic and phototrophic biotechnologies we performed a comprehensive literature review and analysis, focusing on energy production (as kJ per capita per day and as kJ m-3 of wastewater treated), energy consumption, and treatment efficacy. Anaerobic technologies included in this review were the anaerobic baffled reactor (ABR), anaerobic membrane bioreactor (AnMBR), anaerobic fluidized bed reactor (AFB), upflow anaerobic sludge blanket (UASB), anaerobic sequencing batch reactor (ASBR), microbial electrolysis cell (MEC), and microbial fuel cell (MFC). Phototrophic technologies included were the high rate algal pond (HRAP), photobioreactor (PBR), stirred tank reactor, waste stabilization pond (WSP), and algal turf scrubber (ATS). Average energy recovery efficiencies for anaerobic technologies ranged from 1.6% (MFC) to 47.5% (ABR). When including typical percent chemical oxygen demand (COD) removals by each technology, this range would equate to roughly 40-1200 kJ per capita per day or 110-3300 kJ m-3 of treated wastewater. The average bioenergy feedstock production by phototrophic technologies ranged from 1200-4700 kJ per capita per day or 3400-13000 kJ m-3 (exceeding anaerobic technologies and, at times, the energetic content of the influent organic carbon), with usable energy production dependent upon downstream conversion to fuels. Energy consumption analysis showed that energy positive anaerobic wastewater treatment by emerging technologies would require significant reductions of parasitic losses from mechanical mixing and gas sparging. Technology targets and critical barriers for energy-producing technologies are identified, and the role of integrated anaerobic and phototrophic

Engineering of Saccharomyces cerevisiae for the production of fuel ethanol from xylose

NARCIS (Netherlands)

Kuijper, S.M.

2006-01-01

For various reasons mankind is looking for alternatives for fossil fuels. One of these alternatives is ethanol made from plant biomass. However, the plant material when broken down by hydrolysis into its sugar monomers contains a significant amount of xylose, a 5-carbon-sugar or pentose. Contrary to

The impact of anaerobic microorganisms activities in ruminant waste and coal

Science.gov (United States)

Harlia, Ellin; Hamdani, H.; Winantris, Kurnani, Tb. B. A.; Hidayati, Y. A.; Marlina, E. T.; Rahmah, K. N.; Arief, H.; Ridwan, R.; Joni, I. M.

2018-02-01

Ruminant (dairy cattle, beef cattle and buffalo) waste from intensive farming concentrated in highly populated areas when stacked and accumulated in certain heights and in anaerobic condition, may produce Green House Gases (GHGs) which lead to global warming. This condition is generated through fermentation by microorganism contained in livestock waste and biogenic activities on coal. The GHGs include CH4 (methane), CO2 (carbon dioxide) and N2O (nitrous oxide). The GHG emission should be early monitored to minimize greater problems. In the other hand, methane can be utilized as an environmental friendly energy after stored as biogas on digester. The aim of this research is to detect how much GHGs formed from ruminant waste and biogenic activities on coal, which can be utilized as an alternative energy. This research conducted as an explorative study utilizing dairy cattle feces, beef cattle feces, buffalo feces and three types of coal: lignite, bituminous and sub-bituminous, which is separately added into medium 98-5 made from mixture of agar medium and chemical components in powder and crystal form diluted with distilled water and rumen liquid, with six repetitions. Each sample was stored into 250 mL anaerobic digester, observed weekly for period of 4 weeks, analyzed by Gas Chromatography (GC-A14). The result showed that GHGs: CH4, CO2 and N2O were found in all samples. Anticipation of GHGs formation to avoid air pollution is by utilizing livestock waste and coal in aerobic condition or in anaerobic condition through digester.

Novel transporters from Kluyveromyces marxianus and Pichia guilliermondii expressed in Saccharomyces cerevisiae enable growth on L-arabinose and D-xylose.

Science.gov (United States)

Knoshaug, Eric P; Vidgren, Virve; Magalhães, Frederico; Jarvis, Eric E; Franden, Mary Ann; Zhang, Min; Singh, Arjun

2015-10-01

Genes encoding L-arabinose transporters in Kluyveromyces marxianus and Pichia guilliermondii were identified by functional complementation of Saccharomyces cerevisiae whose growth on L-arabinose was dependent on a functioning L-arabinose transporter, or by screening a differential display library, respectively. These transporters also transport D-xylose and were designated KmAXT1 (arabinose-xylose transporter) and PgAXT1, respectively. Transport assays using L-arabinose showed that KmAxt1p has K(m) 263 mM and V(max) 57 nM/mg/min, and PgAxt1p has K(m) 0.13 mM and V(max) 18 nM/mg/min. Glucose, galactose and xylose significantly inhibit L-arabinose transport by both transporters. Transport assays using D-xylose showed that KmAxt1p has K(m) 27 mM and V(max) 3.8 nM/mg/min, and PgAxt1p has K(m) 65 mM and V(max) 8.7 nM/mg/min. Neither transporter is capable of recovering growth on glucose or galactose in a S. cerevisiae strain deleted for hexose and galactose transporters. Transport kinetics of S. cerevisiae Gal2p showed K(m) 371 mM and V(max) 341 nM/mg/min for L-arabinose, and K(m) 25 mM and V(max) 76 nM/mg/min for galactose. Due to the ability of Gal2p and these two newly characterized transporters to transport both L-arabinose and D-xylose, one scenario for the complete usage of biomass-derived pentose sugars would require only the low-affinity, high-throughput transporter Gal2p and one additional high-affinity general pentose transporter, rather than dedicated D-xylose or L-arabinose transporters. Additionally, alignment of these transporters with other characterized pentose transporters provides potential targets for substrate recognition engineering. Copyright © 2015 John Wiley & Sons, Ltd.

On the use of prior information in modelling metabolic utilization of energy in growing pigs

DEFF Research Database (Denmark)

Strathe, Anders Bjerring; Jørgensen, Henry; Fernández, José Adalberto

2011-01-01

Construction of models that provide a realistic representation of metabolic utilization of energy in growing animals tend to be over-parameterized because data generated from individual metabolic studies are often sparse. In the Bayesian framework prior information can enter the data analysis......, PD and LD) made on a given pig at a given time followed a multivariate normal distribution. Two different equation systems were adopted from Strathe et al. (2010), generating the expected values in the multivariate normal distribution. Non-informative prior distributions were assigned for all model......, kp and kf, respectively. Utilizing both sets of priors showed that the maintenance component was sensitive to the statement of prior belief and, hence, that the estimate of 0.91 MJkg0.60d1 (95% CI: 0.78; 1.09) should be interpreted with caution. It was shown that boars were superior in depositing...

Transcriptomic comparison of Aspergillus niger growing on two different sugars reveals coordinated regulation of the secretory pathway

DEFF Research Database (Denmark)

Jørgensen, Thomas R; Goosen, Theo; Hondel, Cees A M J J van den

2009-01-01

BACKGROUND: The filamentous fungus, Aspergillus niger, responds to nutrient availability by modulating secretion of various substrate degrading hydrolases. This ability has made it an important organism in industrial production of secreted glycoproteins. The recent publication of the A. niger...... the physiology and transcriptome of A. niger growing at the same specific growth rate (0.16 h(-1)) on xylose or maltose in carbon-limited chemostat cultures. Transcription profiles were obtained using Affymetrix GeneChip analysis of six replicate cultures for each of the two growth-limiting carbon sources...

A Highly Efficient Xylan-Utilization System in Aspergillus niger An76: A Functional-Proteomics Study

Directory of Open Access Journals (Sweden)

Weili Gong

2018-03-01

Full Text Available Xylan constituted with β-1,4-D-xylose linked backbone and diverse substituted side-chains is the most abundant hemicellulose component of biomass, which can be completely and rapidly degraded into fermentable sugars by Aspergillus niger. This is of great value for obtaining renewable biofuels and biochemicals. To clarify the underlying mechanisms associated with highly efficient xylan degradation, assimilation, and metabolism by A. niger, we utilized functional proteomics to analyze the secreted proteins, sugar transporters, and intracellular proteins of A. niger An76 grown on xylan-based substrates. Results demonstrated that the complete xylanolytic enzyme system required for xylan degradation and composed of diverse isozymes was secreted in a sequential order. Xylan-backbone-degrading enzymes were preferentially induced by xylose or other soluble sugars, which efficiently produced large amounts of xylooligosaccharides (XOS and xylose; however, XOS was more efficient than xylose in triggering the expression of the key transcription activator XlnR, resulting in higher xylanase activity and shortening xylanase-production time. Moreover, the substituted XOS was responsible for improving the abundance of side-chain-degrading enzymes, specific transporters, and key reductases and dehydrogenases in the pentose catabolic pathway. Our findings indicated that industries might be able to improve the species and concentrations of xylan-degrading enzymes and shorten fermentation time by adding abundant intermediate products of natural xylan (XOS to cultures of filamentous fungi.

A Highly Efficient Xylan-Utilization System in Aspergillus niger An76: A Functional-Proteomics Study

Science.gov (United States)

Gong, Weili; Dai, Lin; Zhang, Huaiqiang; Zhang, Lili; Wang, Lushan

2018-01-01

Xylan constituted with β-1,4-D-xylose linked backbone and diverse substituted side-chains is the most abundant hemicellulose component of biomass, which can be completely and rapidly degraded into fermentable sugars by Aspergillus niger. This is of great value for obtaining renewable biofuels and biochemicals. To clarify the underlying mechanisms associated with highly efficient xylan degradation, assimilation, and metabolism by A. niger, we utilized functional proteomics to analyze the secreted proteins, sugar transporters, and intracellular proteins of A. niger An76 grown on xylan-based substrates. Results demonstrated that the complete xylanolytic enzyme system required for xylan degradation and composed of diverse isozymes was secreted in a sequential order. Xylan-backbone-degrading enzymes were preferentially induced by xylose or other soluble sugars, which efficiently produced large amounts of xylooligosaccharides (XOS) and xylose; however, XOS was more efficient than xylose in triggering the expression of the key transcription activator XlnR, resulting in higher xylanase activity and shortening xylanase-production time. Moreover, the substituted XOS was responsible for improving the abundance of side-chain-degrading enzymes, specific transporters, and key reductases and dehydrogenases in the pentose catabolic pathway. Our findings indicated that industries might be able to improve the species and concentrations of xylan-degrading enzymes and shorten fermentation time by adding abundant intermediate products of natural xylan (XOS) to cultures of filamentous fungi. PMID:29623069

A Highly Efficient Xylan-Utilization System in Aspergillus niger An76: A Functional-Proteomics Study.

Science.gov (United States)

Gong, Weili; Dai, Lin; Zhang, Huaiqiang; Zhang, Lili; Wang, Lushan

2018-01-01

Xylan constituted with β-1,4-D-xylose linked backbone and diverse substituted side-chains is the most abundant hemicellulose component of biomass, which can be completely and rapidly degraded into fermentable sugars by Aspergillus niger . This is of great value for obtaining renewable biofuels and biochemicals. To clarify the underlying mechanisms associated with highly efficient xylan degradation, assimilation, and metabolism by A. niger , we utilized functional proteomics to analyze the secreted proteins, sugar transporters, and intracellular proteins of A. niger An76 grown on xylan-based substrates. Results demonstrated that the complete xylanolytic enzyme system required for xylan degradation and composed of diverse isozymes was secreted in a sequential order. Xylan-backbone-degrading enzymes were preferentially induced by xylose or other soluble sugars, which efficiently produced large amounts of xylooligosaccharides (XOS) and xylose; however, XOS was more efficient than xylose in triggering the expression of the key transcription activator XlnR, resulting in higher xylanase activity and shortening xylanase-production time. Moreover, the substituted XOS was responsible for improving the abundance of side-chain-degrading enzymes, specific transporters, and key reductases and dehydrogenases in the pentose catabolic pathway. Our findings indicated that industries might be able to improve the species and concentrations of xylan-degrading enzymes and shorten fermentation time by adding abundant intermediate products of natural xylan (XOS) to cultures of filamentous fungi.

Biphasic single-reactor process for dehydration of xylose and hydrogenation of produced furfural

NARCIS (Netherlands)

Ordomskiy, V.; Schouten, J.C.; Schaaf, van der J.; Nijhuis, T.A.

2013-01-01

The processes of xylose dehydration and the consecutive furfural hydrogenation have been combined in a single biphasic reactor. The dehydration was studied over Amberlyst-15 and the hydrogenation over a hydrophobic Ru/C catalyst. 1-Butanol, 2-methyltetrahydrofuran and cyclohexane were used as

PENGOLAHAN LIMBAH CAIR INDUSTRI FARMASI FORMULASI DENGAN METODE ANAEROB-AEROB DAN ANAEROB-KOAGULASI

Directory of Open Access Journals (Sweden)

Farida Crisnaningtyas

2016-05-01

Full Text Available Studi ini membahas mengenai pengolahan limbah cair industri farmasi dalam skala laboratorium dengan menggunakan konsep anaerob-kimia-fisika dan anaerob-aerob. Proses anaerob dilakukan dengan menggunakan reaktor Upflow Anaerobic Sludge Bed reactor (UASBr pada kisaran OLR (Organic Loading Rate 0,5 – 2 kg COD/m3hari, yang didahului dengan proses aklimatisasi menggunakan substrat gula. Proses anaerob mampu memberikan efisiensi penurunan COD hingga 74%. Keluaran dari proses anaerob diolah lebih lanjut dengan menggunakan dua opsi proses: (1 fisika-kimia, dan (2 aerob. Koagulan alumunium sulfat dan flokulan kationik memberikan efisiensi penurunan COD tertinggi (73% pada kecepatan putaran masing-masing 100 rpm dan 40 rpm. Uji coba aerob dilakukan pada kisaran MLSS antara 4000-5000 mg/L dan mampu memberikan efisiensi penurunan COD hingga 97%. Hasil uji coba menunjukkan bahwa efisiensi penurunan COD total yang dapat dicapai dengan menggunakan teknologi anaerob-aerob adalah 97%, sedangkan kombinasi anaerob-koagulasi-flokulasi hanya mampu menurunkan COD total sebesar 72,53%. Berdasarkan hasil tersebut, kombinasi proses anaerob-aerob merupakan teknologi yang potensial untuk diaplikasikan dalam sistem pengolahan limbah cair industri farmasi. 

Indigenous utilization of termite mounds and their sustainability in a rice growing village of the central plain of Laos.

Science.gov (United States)

Miyagawa, Shuichi; Koyama, Yusaku; Kokubo, Mika; Matsushita, Yuichi; Adachi, Yoshinao; Sivilay, Sengdeaune; Kawakubo, Nobumitsu; Oba, Shinya

2011-08-18

The objective of this study was to investigate the indigenous utilization of termite mounds and termites in a rain-fed rice growing village in the central plain of Laos, where rice production is low and varies year-to-year, and to assess the possibility of sustainable termite mound utilization in the future. This research was carried out from 2007 to 2009. The termites were collected from their mounds and surrounding areas and identified. Twenty villagers were interviewed on their use of termites and their mounds in the village. Sixty-three mounds were measured to determine their dimensions in early March, early July and middle to late November, 2009. Eleven species of Termitidae were recorded during the survey period. It was found that the villagers use termite mounds as fertilizer for growing rice, vegetable beds and charcoal kilns. The villagers collected termites for food and as feed for breeding fish. Over the survey period, 81% of the mounds surveyed increased in volume; however, the volume was estimated to decrease by 0.114 m3 mound(-1) year(-1) on average due to several mounds being completely cut out. It was concluded that current mound utilization by villagers is not sustainable. To ensure sustainable termite utilization in the future, studies should be conducted to enhance factors that promote mound restoration by termites. Furthermore, it will be necessary to improve mound conservation methods used by the villagers after changes in the soil mass of mounds in paddy fields and forests has been measured accurately. The socio-economic factors that affect mound utilization should also be studied.

Influence Of Quinolone Lethality on Irradiated Anaerobic Growth of Escherichia Coli

International Nuclear Information System (INIS)

Ibrahim, I.M.; El-Kabbany, H.M.; El-Esseily, E.SH.

2012-01-01

Bacteriostatic and bactericidal activities were measured with wild type cells and isomerase mutants of Escherichia coli for ciprofloxacin, formation of quinolone-gyrase-DNA complexes, observed as a sodium dodecyl sulfate (SDS) dependent drop in cell lysate viscosity, occurred during aerobic and anaerobic growth and in the presence and in the absence of chloramphenicol. Quinolone activity against Escherichia coli was examined during aerobic growth, aerobic treatment with chloramphenicol, and anaerobic growth. Nalidixic acid, norfloxacin and ciprofloxacin were lethal for cultures growing aerobically, and the bacteriostatic activity of each quinolone was unaffected by anaerobic growth. However, lethal activity was distinct for each quinolone with cells treated aerobically with chloramphenicol or grown anaerobically. Nalidixic acid failed to kill cells under both conditions, norfloxacin killed cells when they were grown anaerobically but not when they were treated with chloramphenicol, ciprofloxacin killed cells under both conditions but required higher concentrations than those required with cells grown aerobically, C-methoxy fluoro quinolone was equally lethal under all conditions. However, lethal chromosome fragmentation, detected as a drop in viscosity in the absence of SDS, was occurred with nalidixic acid treatment only under aerobic conditions in the absence of chloramphenicol, thus, all quinolones tested appeared to form reversible bacteriostatic complexes containing broken DNA during aerobic growth, during anaerobic growth, and when protein synthesis is blocked. The ability to fragment chromosomes rapidly kill cells under these conditions depends on quinolone structure. The radiation of sublethal dose was 3 Gy at rate of 0.6 Gy/min was shown as non-significant result

Systematic strain construction and process development: Xylitol production by Saccharomyces cerevisiae expressing Candida tenuis xylose reductase in wild-type or mutant form.

Science.gov (United States)

Pratter, S M; Eixelsberger, T; Nidetzky, B

2015-12-01

A novel Saccharomyces cerevisiae whole-cell biocatalyst for xylitol production based on Candida tenuis xylose reductase (CtXR) is presented. Six recombinant strains expressing wild-type CtXR or an NADH-specific mutant were constructed and evaluated regarding effects of expression mode, promoter strength, biocatalyst concentration and medium composition. Intracellular XR activities ranged from 0.09 U mgProt(-1) to 1.05 U mgProt(-1) but did not correlate with the strains' xylitol productivities, indicating that other factors limited xylose conversion in the high-activity strains. The CtXR mutant decreased the biocatalyst's performance, suggesting use of the NADPH-preferring wild-type enzyme when (semi-)aerobic conditions are applied. In a bioreactor process, the best-performing strain converted 40 g L(-1) xylose with an initial productivity of 1.16 g L(-1)h(-1) and a xylitol yield of 100%. The obtained results underline the potential of CtXR wild-type for xylose reduction and point out parameters to improve "green" xylitol production. Copyright © 2015 Elsevier Ltd. All rights reserved.

An innovative intermittent-vacuum assisted thermophilic anaerobic digestion process for effective animal manure utilization and treatment.

Science.gov (United States)

Zhang, Renchuan; Anderson, Erik; Addy, Min; Deng, Xiangyuan; Kabir, Fayal; Lu, Qian; Ma, Yiwei; Cheng, Yanling; Liu, Yuhuan; Chen, Paul; Ruan, Roger

2017-11-01

Intermittent-vacuum stripping (IVS) was developed as a pretreatment for thermophilic anaerobic digestion (TAD) to improve methanogenesis and hydrolysis activity through preventing free ammonia and hydrogen sulfide (H 2 S) inhibition from liquid swine manure (LSM). Over 98% of ammonia and 38% organic nitrogen were removed in 60min from 55°C to 85°C with vacuum pressure (from 100.63±3.79mmHg to 360.91±7.39mmHg) at initial pH 10.0 by IVS. Thermophilic methanogenesis and hydrolysis activity of pretreated LSM increased 52.25% (from 11.56±1.75% to 17.60±0.49%) in 25days and 40% (from 10days to 6days) in bio-methane potential assay. Over 80% H 2 S and total nitrogen were removed by IVS assistance, while around 70% nitrogen was recycled as ammonium sulfate. Therefore, IVS-TAD combination could be an effective strategy to improve TAD efficiency, whose elution is more easily utilized in algae cultivation and/or hydroponic system. Copyright © 2017 Elsevier Ltd. All rights reserved.

Biobutanol production by Clostridium acetobutylicum using xylose recovered from birch Kraft black liquor.

Science.gov (United States)

Kudahettige-Nilsson, Rasika L; Helmerius, Jonas; Nilsson, Robert T; Sjöblom, Magnus; Hodge, David B; Rova, Ulrika

2015-01-01

Acetone-butanol-ethanol (ABE) fermentation was studied using acid-hydrolyzed xylan recovered from hardwood Kraft black liquor by CO2 acidification as the only carbon source. Detoxification of hydrolyzate using activated carbon was conducted to evaluate the impact of inhibitor removal and fermentation. Xylose hydrolysis yields as high as 18.4% were demonstrated at the highest severity hydrolysis condition. Detoxification using active carbon was effective for removal of both phenolics (76-81%) and HMF (38-52%). Batch fermentation of the hydrolyzate and semi-defined P2 media resulted in a total solvent yield of 0.12-0.13g/g and 0.34g/g, corresponding to a butanol concentration of 1.8-2.1g/L and 7.3g/L respectively. This work is the first study of a process for the production of a biologically-derived biofuel from hemicelluloses solubilized during Kraft pulping and demonstrates the feasibility of utilizing xylan recovered directly from industrial Kraft pulping liquors as a feedstock for biological production of biofuels such as butanol. Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.

Ethanol production from cellulose, lactose and xylose using yeasts and enzymes. Gewinnung von Ethanol aus Cellulose, Lactose, und Xylose mit Hilfe von Hefen und Enzymen

Energy Technology Data Exchange (ETDEWEB)

Schwank, U

1986-07-03

Experiments with mixtures of whey and corn showed that more than 85% of the lactose was degraded into ethanol. The applicability of cellulose was investigated by means of potatoes. Cellulase is inhibited by glucose, which is a fermentation intermediate, as well as by the end product ethanol. A cellulase inhibitor in potatoes was detected and stabilized; this inhibitor could be degraded into neutral components by a suitable enzyme. Saccharification and fermentation experiments showed that the cellulose fraction of potatoes can be reduced efficiently. The effects of non-enzymatic pretreatment on enzymatic degradation of cellulose, combined with fermentation of the degradation products, are illustrated by the example of cellulose treated with acid and alkaline substances. A continuous fermentation system was developed from which the ethanol is withdrawn in vapour form. The system made better use of the cellulase activity and increased the efficiency of a xylose-fermenting yeast. The new method is compared with batch experiments in order to assess its efficiency. The advantages of the continuous process are proved for two yeasts of the species Pachysolu and Pichia. Specific fermentation rates up to 0.08 g/(g x h) and fermentation yields up to 0.42 g ethanol/g xylose were achieved with Pichia stipitis.

Continuous biohydrogen production from waste bread by anaerobic sludge.

Science.gov (United States)

Han, Wei; Huang, Jingang; Zhao, Hongting; Li, Yongfeng

2016-07-01

In this study, continuous biohydrogen production from waste bread by anaerobic sludge was performed. The waste bread was first hydrolyzed by the crude enzymes which were generated by Aspergillus awamori and Aspergillus oryzae via solid-state fermentation. It was observed that 49.78g/L glucose and 284.12mg/L free amino nitrogen could be produced with waste bread mass ratio of 15% (w/v). The waste bread hydrolysate was then used for biohydrogen production by anaerobic sludge in a continuous stirred tank reactor (CSTR). The optimal hydrogen production rate of 7.4L/(Ld) was achieved at chemical oxygen demand (COD) of 6000mg/L. According to the results obtained from this study, 1g waste bread could generate 0.332g glucose which could be further utilized to produce 109.5mL hydrogen. This is the first study which reports continuous biohydrogen production from waste bread by anaerobic sludge. Copyright © 2016 Elsevier Ltd. All rights reserved.

Enrichment of Thermophilic Syntrophic Anaerobic Glutamate-Degrading Consortia using a Dialysis Membrane Reactor

NARCIS (Netherlands)

Plugge, C.M.; Stams, A.J.M.

2002-01-01

A dialysis cultivation system was used to enrich slow-growing moderately thermophilic anaerobic bacteria at high cell densities. Bicarbonate buffered mineral salts medium with 5 mM glutamate as the sole carbon and energy source was used and the incubation temperature was 55 degrees C. The reactor

Dehydration of D-xylose over SiO{sub 2}-Al{sub 2}O{sub 3} catalyst: Perspective on the pathways for condensed products

Energy Technology Data Exchange (ETDEWEB)

You, Su Jin; Park, Eun Duck; Park, Myung-June [Ajou University, Suwon (Korea, Republic of)

2016-03-15

This work addresses the kinetic mechanism for the dehydration of D-xylose over the SiO{sub 2}-Al{sub 2}O{sub 3} solid catalyst, where the formation of condensed products is included in addition to the production of furfural and its decomposition. The kinetic modeling and parametric sensitivity show that the isomerization of D-xylose takes place in the early stages of the reaction, followed by the dehydration of isomers. Accordingly, the homogeneous polymerization of isomers is found to be dominant. The developed model is used to evaluate the effects of operating conditions on the catalytic performance; high temperature and D-xylose concentration guarantee high furfural yield.

Retention and transport of an anaerobic trichloroethene dechlorinating microbial culture in anaerobic porous media.

Science.gov (United States)

Zhang, Huixin; Ulrich, Ania C; Liu, Yang

2015-06-01

The influence of solution chemistry on microbial transport was examined using the strictly anaerobic trichloroethene (TCE) bioaugmentation culture KB-1(®). A column was employed to determine transport behaviors and deposition kinetics of three distinct functional species in KB-1(®), Dehalococcoides, Geobacter, and Methanomethylovorans, over a range of ionic strengths under a well-controlled anaerobic condition. A quantitative polymerase chain reaction (qPCR) was utilized to enumerate cell concentration and complementary techniques were implemented to evaluate cell surface electrokinetic potentials. Solution chemistry was found to positively affect the deposition rates, which was consistent with calculated Derjaguin-Landau-Verwey-Overbeek (DLVO) interaction energies. Retained microbial profiles showed spatially constant colloid deposition rate coefficients, in agreement with classical colloid filtration theory (CFT). It was interesting to note that the three KB-1(®) species displayed similar transport and retention behaviors under the defined experimental conditions despite their different cell electrokinetic properties. A deeper analysis of cell characteristics showed that factors, such as cell size and shape, concentration, and motility were involved in determining adhesion behavior. Copyright © 2015 Elsevier B.V. All rights reserved.

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.

Anaerobic degradation of aircraft deicing fluid (ADF) in upflow anaerobic sludge blanket (UASB) reactors and the fate of ADF additives

Science.gov (United States)

Pham, Thi Tham

2002-11-01

effects on acidogenesis and methanogenesis at the concentration levels studied. A significant inhibition of acetoclastic activity was observed for NP at 100 mg/L, with acetic acid consumption rate at 38% of that for controls. No evidence for anaerobic degradation of benzotriazole and its derivatives was observed; however, both batch and continuous experiments suggested that anaerobic degradation of NP occurred. Kinetic analysis of operational data obtained for the anaerobic treatment of ADF in UASB reactors indicated that the substrate utilization rate was independent of the reactor biomass concentration. The maximum rate of substrate utilization and the half-velocity constants for ADF treatment were 28.4 g COD/L/d and 648 mg COD/L, respectively. For 1.2% ADF, the biomass yield and endogenous decay coefficients were 0.027 g VSS/g COD and 0.012 d-1 , respectively.

Energy positive domestic wastewater treatment: the roles of anaerobic and phototrophic technologies

KAUST Repository

Shoener, B. D.; Bradley, I. M.; Cusick, R. D.; Guest, J. S.

2014-01-01

The negative energy balance of wastewater treatment could be reversed if anaerobic technologies were implemented for organic carbon oxidation and phototrophic technologies were utilized for nutrient recovery. To characterize the potential for energy

Quantitative investigations of xylose and arabinose substituents in hydroxypropylated and hydroxyvinylethylated arabinoxylans.

Science.gov (United States)

Lorenz, Dominic; Knöpfle, Anna; Akil, Youssef; Saake, Bodo

2017-11-01

The chemical structures obtained by the modification of arabinoxylans with the cyclic carbonates propylene carbonate (PC) and 4-vinyl-1,3-dioxolan-2-one (VEC) with varying degrees of substitution were investigated. Therefore, a new analytical method was developed that is based on a microwave-assisted hydrolysis of the polysaccharides with trifluoroacetic acid and the reductive amination with 2-aminobenzoic acid. The peak assignment was achieved by HPLC-MS and the carbohydrate derivatives were quantified by HPLC-fluorescence. The obtained maximum molar substitution of PC-derivatized xylan (X HP ) was 1.8; the molar substitution of VEC-derivatized xylan (X HVE ) was 2.3. Investigations of xylose and arabinose based mono- and disubstituted derivatives revealed a preferred reaction of the cyclic carbonates with arabinose. Conversion rates were up to 2.4 times higher for monosubstitution and up to 3.0 times for disubstitution compared to xylose. Furthermore, the reaction with VEC was preferred due to higher reactivity of the newly introduced side chains. Copyright © 2017 Elsevier Ltd. All rights reserved.

Genes related to xylose fermentation and methods of using same for enhanced biofuel production

Science.gov (United States)

Wohlbach, Dana J.; Gasch, Audrey P.

2014-08-05

The present invention provides isolated gene sequences involved in xylose fermentation and related recombinant yeast which are useful in methods of enhanced biofuel production, particularly ethanol production. Methods of bioengineering recombinant yeast useful for biofuel production are also provided.

Molecular analysis of the biomass of a fluidized bed reactor treating synthetic vinasse at anaerobic and micro-aerobic conditions.

Science.gov (United States)

Rodríguez, Elisa; Lopes, Alexandre; Fdz-Polanco, María; Stams, Alfons J M; García-Encina, Pedro A

2012-03-01

The microbial communities (Bacteria and Archaea) established in an anaerobic fluidized bed reactor used to treat synthetic vinasse (betaine, glucose, acetate, propionate, and butyrate) were characterized by denaturing gradient gel electrophoresis (DGGE) and phylogenetic analysis. This study was focused on the competitive and syntrophic interactions between the different microbial groups at varying influent substrate to sulfate ratios of 8, 4, and 2 and anaerobic or micro-aerobic conditions. Acetogens detected along the anaerobic phases at substrate to sulfate ratios of 8 and 4 seemed to be mainly involved in the fermentation of glucose and betaine, but they were substituted by other sugar or betaine degraders after oxygen application. Typical fatty acid degraders that grow in syntrophy with methanogens were not detected during the entire reactor run. Likely, sugar and betaine degraders outnumbered them in the DGGE analysis. The detected sulfate-reducing bacteria (SRB) belonged to the hydrogen-utilizing Desulfovibrio. The introduction of oxygen led to the formation of elemental sulfur (S(0)) and probably other sulfur compounds by sulfide-oxidizing bacteria (γ-Proteobacteria). It is likely that the sulfur intermediates produced from sulfide oxidation were used by SRB and other microorganisms as electron acceptors, as was supported by the detection of the sulfur respiring Wolinella succinogenes. Within the Archaea population, members of Methanomethylovorans and Methanosaeta were detected throughout the entire reactor operation. Hydrogenotrophic methanogens mainly belonging to the genus Methanobacterium were detected at the highest substrate to sulfate ratio but rapidly disappeared by increasing the sulfate concentration.

Sulfur and Oxygen Isotope Fractionation During Bacterial Sulfur Disproportionation Under Anaerobic Haloalkaline Conditions

NARCIS (Netherlands)

Poser, Alexander; Vogt, Carsten; Knöller, Kay; Sorokin, Dimitry Y.; Finster, Kai W.; Richnow, Hans H.

2016-01-01

Sulfur and oxygen isotope fractionation of elemental sulfur disproportionation at anaerobic haloalkaline conditions was evaluated for the first time. Isotope enrichment factors of the strains Desulfurivibrio alkaliphilus and Dethiobacter alkaliphilus growing at pH 9 or 10 were −0.9‰ to −1‰ for

Ethanol fermentation by xylose-assimilating Saccharomyces cerevisiae using sugars in a rice straw liquid hydrolysate concentrated by nanofiltration.

Science.gov (United States)

Sasaki, Kengo; Sasaki, Daisuke; Sakihama, Yuri; Teramura, Hiroshi; Yamada, Ryosuke; Hasunuma, Tomohisa; Ogino, Chiaki; Kondo, Akihiko

2013-11-01

Concentrating sugars using membrane separation, followed by ethanol fermentation by recombinant xylose-assimilating Saccharomyces cerevisiae, is an attractive technology. Three nanofiltration membranes (NTR-729HF, NTR-7250, and ESNA3) were effective in concentrating glucose, fructose, and sucrose from dilute molasses solution and no permeation of sucrose. The separation factors of acetate, formate, furfural, and 5-hydroxymethyl furfural, which were produced by dilute acid pretreatment of rice straw, over glucose after passage through these three membranes were 3.37-11.22, 4.71-20.27, 4.32-16.45, and 4.05-16.84, respectively, at pH 5.0, an applied pressure of 1.5 or 2.0 MPa, and 25 °C. The separation factors of these fermentation inhibitors over xylose were infinite, as there was no permeation of xylose. Ethanol production from approximately two-times concentrated liquid hydrolysate using recombinant S. cerevisiae was double (5.34-6.44 g L(-1)) that compared with fermentation of liquid hydrolysate before membrane separation (2.75 g L(-1)). Copyright © 2013 Elsevier Ltd. All rights reserved.

Indigenous utilization of termite mounds and their sustainability in a rice growing village of the central plain of Laos

Directory of Open Access Journals (Sweden)

Sivilay Sengdeaune

2011-08-01

Full Text Available Abstract Background The objective of this study was to investigate the indigenous utilization of termite mounds and termites in a rain-fed rice growing village in the central plain of Laos, where rice production is low and varies year-to-year, and to assess the possibility of sustainable termite mound utilization in the future. This research was carried out from 2007 to 2009. Methods The termites were collected from their mounds and surrounding areas and identified. Twenty villagers were interviewed on their use of termites and their mounds in the village. Sixty-three mounds were measured to determine their dimensions in early March, early July and middle to late November, 2009. Results Eleven species of Termitidae were recorded during the survey period. It was found that the villagers use termite mounds as fertilizer for growing rice, vegetable beds and charcoal kilns. The villagers collected termites for food and as feed for breeding fish. Over the survey period, 81% of the mounds surveyed increased in volume; however, the volume was estimated to decrease by 0.114 m3 mound-1 year-1 on average due to several mounds being completely cut out. Conclusion It was concluded that current mound utilization by villagers is not sustainable. To ensure sustainable termite utilization in the future, studies should be conducted to enhance factors that promote mound restoration by termites. Furthermore, it will be necessary to improve mound conservation methods used by the villagers after changes in the soil mass of mounds in paddy fields and forests has been measured accurately. The socio-economic factors that affect mound utilization should also be studied.

Sensitive and selective culture medium for detection of environmental Clostridium difficile isolates without requirement for anaerobic culture conditions.

Science.gov (United States)

Cadnum, Jennifer L; Hurless, Kelly N; Deshpande, Abhishek; Nerandzic, Michelle M; Kundrapu, Sirisha; Donskey, Curtis J

2014-09-01

Effective and easy-to-use methods for detecting Clostridium difficile spore contamination would be useful for identifying environmental reservoirs and monitoring the effectiveness of room disinfection. Culture-based detection methods are sensitive for detecting C. difficile, but their utility is limited due to the requirement of anaerobic culture conditions and microbiological expertise. We developed a low-cost selective broth medium containing thioglycolic acid and l-cystine, termed C. difficile brucella broth with thioglycolic acid and l-cystine (CDBB-TC), for the detection of C. difficile from environmental specimens under aerobic culture conditions. The sensitivity and specificity of CDBB-TC (under aerobic culture conditions) were compared to those of CDBB (under anaerobic culture conditions) for the recovery of C. difficile from swabs collected from hospital room surfaces. CDBB-TC was significantly more sensitive than CDBB for recovering environmental C. difficile (36/41 [88%] versus 21/41 [51%], respectively; P = 0.006). C. difficile latex agglutination, an enzyme immunoassay for toxins A and B or glutamate dehydrogenase, and a PCR for toxin B genes were all effective as confirmatory tests. For 477 total environmental cultures, the specificity of CDBB-TC versus that of CDBB based upon false-positive yellow-color development of the medium without recovery of C. difficile was 100% (0 false-positive results) versus 96% (18 false-positive results), respectively. False-positive cultures for CDBB were attributable to the growth of anaerobic non-C. difficile organisms that did not grow in CDBB-TC. Our results suggest that CDBB-TC provides a sensitive and selective medium for the recovery of C. difficile organisms from environmental samples, without the need for anaerobic culture conditions. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Accelerated anaerobic hydrolysis rates under a combination of intermittent aeration and anaerobic conditions

DEFF Research Database (Denmark)

Jensen, T. R.; Lastra Milone, T.; Petersen, G.

2017-01-01

Anaerobic hydrolysis in activated return sludge was investigated in laboratory scale experiments to find if intermittent aeration would accelerate anaerobic hydrolysis rates compared to anaerobic hydrolysis rates under strict anaerobic conditions. The intermittent reactors were set up in a 240 h...... for calculating hydrolysis rates based on soluble COD were compared. Two-way ANOVA with the Bonferroni post-test was performed in order to register any significant difference between reactors with intermittent aeration and strictly anaerobic conditions respectively. The experiment demonstrated a statistically...... significant difference in favor of the reactors with intermittent aeration showing a tendency towards accelerated anaerobic hydrolysis rates due to application of intermittent aeration. The conclusion of the work is thus that intermittent aeration applied in the activated return sludge process (ARP) can...

Co-fermentation of the main sugar types from a beechwood organosolv hydrolysate by several strains of Bacillus coagulans results in effective lactic acid production

Directory of Open Access Journals (Sweden)

Robert Glaser

2018-06-01

Full Text Available Bacillus coagulans is an interesting facultative anaerobic microorganism for biotechnological production of lactic acid that arouses interest. To determine the efficiency of biotechnological production of lactic acid from lignocellulosic feedstock hydrolysates, five Bacillus coagulans strains were grown in lignocellulose organosolv hydrolysate from ethanol/water-pulped beechwood. Parameter estimation based on a Monod-type model was used to derive the basic key parameters for a performance evaluation of the batch process. Three of the Bacillus coagulans strains, including DSM No. 2314, were able to produce lactate, primarily via uptake of glucose and xylose. Two other strains were identified as having the ability of utilizing cellobiose to a high degree, but they also had a lower affinity to xylose. The lactate yield concentration varied from 79.4 ± 2.1 g/L to 93.7 ± 1.4 g/L (85.4 ± 4.7 % of consumed carbohydrates from the diluted organosolv hydrolysate.

The Effect of Anaerobic and Aerobic Fish Sludge Supernatant on Hydroponic Lettuce

Directory of Open Access Journals (Sweden)

Simon Goddek

2016-06-01

Full Text Available The mobilization of nutrients from fish sludge (i.e., feces and uneaten feed plays a key role in optimizing the resource utilization and thus in improving the sustainability of aquaponic systems. While several studies have documented the aerobic and anaerobic digestion performance of aquaculture sludge, the impact of the digestate on plant growth has yet to be understood. The present study examines the impact of either an aerobic or an anaerobic digestion effluent on lettuce plant growth, by enriching a mixture of aquaculture and tap water with supernatants from both aerobic and anaerobic batch reactors. The lettuce plants grown in the hydroponic system supplied with supernatant from an anaerobic reactor had significantly better performance with respect to weight gain than both, those in the system where supernatant from the aerobic reactor was added, as well as the control system. It can be hypothesized that this effect was caused by the presence of NH4+ as well as dissolved organic matter, plant growth promoting rhizobacteria and fungi, and humic acid, which are predominantly present in anaerobic effluents. This study should therefore be of value to researchers and practitioners wishing to further develop sludge remineralization in aquaponic systems.

Differential RNA-seq, Multi-Network Analysis and Metabolic Regulation Analysis of Kluyveromyces marxianus Reveals a Compartmentalised Response to Xylose.

Directory of Open Access Journals (Sweden)

Du Toit W P Schabort

Full Text Available We investigated the transcriptomic response of a new strain of the yeast Kluyveromyces marxianus, in glucose and xylose media using RNA-seq. The data were explored in a number of innovative ways using a variety of networks types, pathway maps, enrichment statistics, reporter metabolites and a flux simulation model, revealing different aspects of the genome-scale response in an integrative systems biology manner. The importance of the subcellular localisation in the transcriptomic response is emphasised here, revealing new insights. As was previously reported by others using a rich medium, we show that peroxisomal fatty acid catabolism was dramatically up-regulated in a defined xylose mineral medium without fatty acids, along with mechanisms to activate fatty acids and transfer products of β-oxidation to the mitochondria. Notably, we observed a strong up-regulation of the 2-methylcitrate pathway, supporting capacity for odd-chain fatty acid catabolism. Next we asked which pathways would respond to the additional requirement for NADPH for xylose utilisation, and rationalised the unexpected results using simulations with Flux Balance Analysis. On a fundamental level, we investigated the contribution of the hierarchical and metabolic regulation levels to the regulation of metabolic fluxes. Metabolic regulation analysis suggested that genetic level regulation plays a major role in regulating metabolic fluxes in adaptation to xylose, even for the high capacity reactions, which is unexpected. In addition, isozyme switching may play an important role in re-routing of metabolic fluxes in subcellular compartments in K. marxianus.

Fuel gas production by anaerobic digestion of kelp

Energy Technology Data Exchange (ETDEWEB)

Troiano, R.A. (Dynatech R/D Co., Cambridge, MA); Wise, D.L.; Augenstein, D.C.; Kispert, R.G.; Cooney, C.L.

1976-12-01

The purpose of the experimental program was to explore the feasibility of the anaerobic digestion of kelp to produce methane. Experiments were carried out with freshly harvested U.S. East Coast kelp, Laminaria saccharina. The use for fuel conversion of the rapidly growing U.S. West Coast kelp, the so-called ''giant kelp,'' Macrocystis pyrifera, has been elsewhere. L. saccharina is similar to M. pyrifera in physical structure as well as chemical composition. Both are brown algae (phaeophyta) of the order Laminariales (kelp). Their principal products of photosynthesis are the sugar alcohol, mannitol, and the polysaccharide, laminarin. The cell walls are composed mostly of algin with some cellulose and fucoidin (a phycocolloid-like algin) and the brown color is due to fucoxanthin pigment. It was anticipated that all these constituents of kelp would be subject to anaerobic digestion. The digester operation, alkali pretreatment of kelp, and a comparison of kelp digestion with other substrates are discussed.

Comparative Evaluation of Anaerobic Bacterial Communities Associated with Roots of Submerged Macrophytes Growing in Marine or Brackish Water Sediments

Science.gov (United States)

Sediment microbial communities are important for seagrass growth and carbon cycling, however relatively few studies have addressed the composition of prokaryotic communities in seagrass bed sediments. Selective media were used enumerate culturable anaerobic bacteria associated ...

Xylose fermentation to biofuels (hydrogen and ethanol) by extreme thermophilic (70 C) mixed culture

DEFF Research Database (Denmark)

Chenxi, Zhao; Karakashev, Dimitar Borisov; Lu, W.

2010-01-01

-xylose corresponding to 55% of the theoretical hydrogen yield based on acetate metabolic pathway. An empirical model was established to reveal the quantitative effect of factors significant for biohydrogen (quadratic model) production and for bioethanol (linear model) production. Changes in hydrogen/ethanol yields...

Improvement on D-xylose to Xylitol Biotransformation by Candida guilliermondii Using Cells Permeabilized with Triton X-100 and Selected Process Conditions.

Science.gov (United States)

Cortez, Daniela Vieira; Mussatto, Solange I; Roberto, Inês Conceição

2016-11-01

Cells of Candida guilliermondii permeabilized with Triton X-100 were able to efficiently produce xylitol from a medium composed only by D-xylose and MgCl 2 ·6H 2 O in potassium phosphate buffer, at 35 °C and pH 6.5. Under these conditions, the results were similar to those obtained when cofactor and co-substrate or nutrients were added to the medium (about 95 % D-xylose was assimilated producing 42 g/L of xylitol, corresponding to 0.80 g/g yield and 2.65 g/L h volumetric productivity). Furthermore, the permeabilized cells kept the D-xylose assimilation in about 90 % and the xylitol production in approx. 40 g/L during three bioconversion cycles of 16 h each. These values are highly relevant when compared to others reported in the literature using enzyme technology and fermentative process, thereby demonstrating the effectiveness of the proposed method. The present study reveals that the use of permeabilized cells is an interesting alternative to obtain high xylitol productivity using low cost medium formulation. This approach may allow the future development of xylitol production from xylose present in lignocellulosic biomass, with additional potential for implementation in biorefinery strategies.

Shotgun proteomics of Aspergillus niger microsomes upon D-xylose induction.

Science.gov (United States)

Ferreira de Oliveira, José Miguel P; van Passel, Mark W J; Schaap, Peter J; de Graaff, Leo H

2010-07-01

Protein secretion plays an eminent role in cell maintenance and adaptation to the extracellular environment of microorganisms. Although protein secretion is an extremely efficient process in filamentous fungi, the mechanisms underlying protein secretion have remained largely uncharacterized in these organisms. In this study, we analyzed the effects of the d-xylose induction of cellulase and hemicellulase enzyme secretion on the protein composition of secretory organelles in Aspergillus niger. We aimed to systematically identify the components involved in the secretion of these enzymes via mass spectrometry of enriched subcellular microsomal fractions. Under each condition, fractions enriched for secretory organelles were processed for tandem mass spectrometry, resulting in the identification of peptides that originate from 1,081 proteins, 254 of which-many of them hypothetical proteins-were predicted to play direct roles in the secretory pathway. d-Xylose induction led to an increase in specific small GTPases known to be associated with polarized growth, exocytosis, and endocytosis. Moreover, the endoplasmic-reticulum-associated degradation (ERAD) components Cdc48 and all 14 of the 20S proteasomal subunits were recruited to the secretory organelles. In conclusion, induction of extracellular enzymes results in specific changes in the secretory subproteome of A. niger, and the most prominent change found in this study was the recruitment of the 20S proteasomal subunits to the secretory organelles.

Energy self-supply of large abattoir by sustainable waste utilization based on anaerobic mono-digestion

International Nuclear Information System (INIS)

Ortner, Markus; Wöss, David; Schumergruber, Alexander; Pröll, Tobias; Fuchs, Werner

2015-01-01

Highlights: • Successful implementation of a new waste and energy concept to large size abattoir. • 85% of slaughterhouse waste accumulated converted to energy by anaerobic digestion. • Coverage of abattoirs’ electrical and thermal energy demand between 50% and 60%. • Reduction of main energy and disposal cost by 63%. • Reduction of greenhouse gas emissions by 79%. - Abstract: Abattoirs have a large number of energy intensive processes. Beside energy supply, disposal costs of animal by-products (ABP) are the main relevant cost drivers. In this study, successful implementation of a new waste and energy management system based on anaerobic digestion is described. Several limitations and technical challenges regarding the anaerobic digestion of the protein rich waste material had to be overcome. The most significant problems were process imbalances such as foaming and floatation as well as high accumulation of volatile fatty acids and low biogas yields caused by lack of essential microelements, high ammonia concentrations and fluctuation in operation temperature. Ultimately, 85% of the waste accumulated during the slaughter process is converted into 2700 MW h thermal and 3200 MW h electrical energy in a biogas combined heat and power (CHP) plant. The thermal energy is optimally integrated into the production process by means of a stratified heat buffer. The energy generated by the biogas CHP-plant can cover a significant share of the energy requirement of the abattoir corresponding to 50% of heat and 60% of electric demand, respectively. In terms of annual cost for energy supply and waste disposal a reduction of 63% from 1.4 Mio € to about 0.5 Mio € could be achieved with the new system. The payback period of the whole investment is approximately 9 years. Beside the economic benefits also the positive environmental impact should be highlighted: a 79% reduction of greenhouse gas emissions from 4.5 Mio kg CO 2 to 0.9 Mio kg CO 2 annually was achieved

Anaerobic biodegradability of macropollutants

DEFF Research Database (Denmark)

Angelidaki, Irini

2002-01-01

A variety of test procedures for determination of anaerobic biodegradability has been reported. This paper reviews the methods developed for determination of anaerobic biodegradability of macro-pollutants. Anaerobic biodegradability of micro-pollutants is not included. Furthermore, factors...

Biochar-mediated reductions in greenhouse gas emissions from soil amended with anaerobic digestates

International Nuclear Information System (INIS)

Martin, Sarah L.; Clarke, Michèle L.; Othman, Mukhrizah; Ramsden, Stephen J.; West, Helen M.

2015-01-01

This investigation examines nitrous oxide (N 2 O) fluxes from soil with simultaneous amendments of anaerobic digestates and biochar. The main source of anthropogenic emissions of N 2 O is agriculture and in particular, manure and slurry application to fields. Anaerobic digestates are increasingly used as a fertiliser and interest is growing in their potential as sources of N 2 O via nitrification and denitrification. Biochar is a stable product of pyrolysis and may affect soil properties such as cation exchange capacity and water holding capacity. Whilst work has been conducted on the effects of biochar amendment on N 2 O emissions in soils fertilised with mineral fertilisers and raw animal manures, little work to date has focused on the effects of biochar on nitrogen transformations within soil amended with anaerobic digestates. The aim of the current investigation was to quantify the effects of biochar application on ammonification, nitrification and N 2 O fluxes within soil amended with three anaerobic digestates derived from different feedstocks. A factorial experiment was undertaken in which a sandy loam soil (Dunnington Heath series) was either left untreated, or amended with three different anaerobic digestates and one of three biochar treatments; 0%, 1% or 3%. Nitrous oxide emissions were greatest from soil amended with anaerobic digestate originating from a maize feedstock. Biochar amendment reduced N 2 O emissions from all treatments, with the greatest effect observed in treatments with maximum emissions. The degree of N 2 O production and efficacy of biochar amelioration of gas emissions is discussed in context of soil microbial biomass and soil available carbon. - Highlights: • Nitrous oxide was emitted from anaerobic digestates applied to soil. • Simultaneous amendment of soil with biochar and anaerobic digestate reduced N 2 O emissions. • Soil nitrate accumulation occurred but was digestate dependent

Isolation from estuarine sediments of a Desulfovibrio strain which can grow on lactate coupled to the reductive dehalogenation of 2,4,6-tribromophenol

Energy Technology Data Exchange (ETDEWEB)

Boyle, A.W.; Phelps, C.D.; Young, L.Y. [Rutgers-The State Univ. of New Jersey, New Brunswick, NJ (United States). Biotechnology Center for Agriculture and the Environment

1999-03-01

Strain TBP-1, an anaerobic bacterium capable of reductively dehalogenating 2,4,6-tribromophenol to phenol, was isolated from estuarine sediments of the Arthur Kill in the New York/New Jersey harbor. It is a gram-negative, motile, vibrio-shaped, obligate anaerobe which grows on lactate, pyruvate, hydrogen, and fumarate when provided sulfate as an electron acceptor. The organism accumulates acetate when grown on lactate and sulfate, contains desulfoviridin, and will not grow in the absence of NaCl. It will not utilize acetate, succinate, propionate, or butyrate for growth via sulfate reduction. When supplied with lactate as an electron donor, strain TBP-1 will utilize sulfate, sulfite, sulfur, and thiosulfate for growth but not nitrate, fumarate, or acrylate. This organism debrominates 2-, 4-, 2,4-, 2,6-, and 2,4,6-bromophenol but not 3- or 2,3-bromophenol or monobrominated benzoates. It will not dehalogenate monochlorinated, fluorinated, or iodinated phenols or chlorinated benzoates. Together with its physiological characteristics, its 16S rRNA gene sequence places it in the genus Desulfovibrio. The average growth yield of strain TBP-1 grown on a defined medium supplemented with lactate and 2,4,6-bromophenol is 3.71 mg of protein/mmol of phenol produced, and the yield was 1.42 mg of protein/mmol of phenol produced when 40bromophenol was the electron acceptor. Average growth yields for Desulfovibrio sp. strain TBP-1 grown with 2,4,6-bromophenol, 4-bromophenol, or sulfate are 0.62, 0.71, and 1.07, respectively. Growth did not occur when either lactate or 2,4,6-bromophenol was omitted from the growth medium. These results indicate that Desulfovibrio sp. strain TBP-1 is capable of growth via halorespiration.

Anaerobic Digestion and its Applications

Science.gov (United States)

Anaerobic digestion is a natural biological process. The initials "AD" may refer to the process of anaerobic digestion, or the built systems of anaerobic digesters. While there are many kinds of digesters, the biology is basically the same for all. Anaerobic digesters are built...

Effect of Addition of High Strength Food Wastes on Anaerobic Digestion of Sewage Sludge

OpenAIRE

Vaidya, Ramola Vinay

2015-01-01

Anaerobic co-digestion of municipal sludge and food wastes high in chemical oxygen demand (COD) has been an area of interest for waste water treatment facilities looking to increase methane production, and at the same time, dispose of the wastes and increase the revenue. However, addition of food wastes containing fats, oils and grease (FOG) to the conventional anaerobic digestion process can be difficult and pose challenges to utilities. Incorporating these wastes into the treatment plants c...

The use of a thermotolerant fermentative Kluyveromyces marxianus IMB3 yeast strain for ethanol production

Energy Technology Data Exchange (ETDEWEB)

Banat, I.M. [Univ. of the United Arab Emirates, Al-Ain (United Arab Emirates). Dept. of Biolology; Singh, D. [Haryana Agriculture Univ., Hisar (India). Dept. of Microbiology; Marchant, R. [Ulster Univ. (United Kingdom). School of Applied Biological and Chemical Sciences

1996-12-31

An investigation was carried out on the growth and ethanol production of a novel thermotolerant ethanol-producing Kluyveromyces marxianus IMB3 yeast strain. It grew aerobically on glucose, lactose, cellobiose, xylose and whey permeate and fermented all the above carbon sources to ethanol at 45 C. This strain was capable of growing under anaerobic chemostat fermentation conditions at 45 C and a dilution rate of 0.15 h{sup -1} and produced {<=}0.9 g/l biomass and 1.8% (v/v) ethanol. An increase in biomass (up to 10.0 g/l) and ethanol (up to 4.3% v/v at 45 C and 7.7% v/v at 40 C) were achieved by applying a continuous two-stage fermentation in sequence (one aerobic and one anerobic stage) or a two-stage anaerobic fermentation with cell recycling. Potential applications, involving alcohol production systems, for use in dairy and wood related industries, were discussed. (orig.)

The fate of methanol in anaerobic bioreactors

OpenAIRE

Florencio, L.

1994-01-01

Methanol is an important component of certain industrial wastewaters. In anaerobic environments, methanol can be utilized by methanogens and acetogens. In wastewater treatment plants, the conversion of methanol into methane is preferred because this conversion is responsible for chemical oxygen demand (COD) removal, whereas with the formation of volatile fatty acids (VFA) little COD removal is achieved. Moreover, the accumulation of VFA can lead to reactor instability due to pH drops...

Morphology and physiology of the dimorphic fungus Mucor circinelloides (syn. M. racemosus) during anaerobic growth

DEFF Research Database (Denmark)

Lübbehüsen, Tina Louise; Nielsen, Jens; Mcintyre, Mhairi

2003-01-01

The dimorphic Mucor circinelloides requires an anaerobic atmosphere and the presence of 30% CO2 to grow as a multipolar budding yeast, otherwise hyphal growth predominates. Establishing other means to control the morphology would be a distinct advantage in the development of a fermentation process...

Systems biology and pathway engineering enable Saccharomyces cerevisiae to utilize C-5 and C-6 sugars simultaneously for cellulosic ethanol production

Science.gov (United States)

Saccharomyces cerevisiae is a traditional industrial workhorse for ethanol production. However, conventional ethanologenic yeast is superior in fermentation of hexose sugars (C-6) such as glucose but unable to utilize pentose sugars (C-5) such as xylose richly embedded in lignocellulosic biomass. In...

Optimization of furfural production from D-xylose with formic acid as catalyst in a reactive extraction system.

Science.gov (United States)

Yang, Wandian; Li, Pingli; Bo, Dechen; Chang, Heying; Wang, Xiaowei; Zhu, Tao

2013-04-01

Furfural is one of the most promising platform chemicals derived from biomass. In this study, response surface methodology (RSM) was utilized to determine four important parameters including reaction temperature (170-210°C), formic acid concentration (5-25 g/L), o-nitrotoluene volume percentage (20-80 vt.%), and residence time (40-200 min). The maximum furfural yield of 74% and selectivity of 86% were achieved at 190°C for 20 g/L formic acid concentration and 75 vt.% o-nitrotoluene by 75 min. The high boiling solvent, o-nitrotoluene, was recommended as extraction solvent in a reactive extraction system to obtain high furfural yield and reduce furfural-solvent separation costs. Although the addition of halides to the xylose solutions enhanced the furfural yield and selectivity, the concentration of halides was not an important factor on the furfural yield and selectivity. Copyright © 2013 Elsevier Ltd. All rights reserved.

Anaerobic Thermophiles

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Francesco Canganella

2014-02-01

Full Text Available The term “extremophile” was introduced to describe any organism capable of living and growing under extreme conditions. With the further development of studies on microbial ecology and taxonomy, a variety of “extreme” environments have been found and an increasing number of extremophiles are being described. Extremophiles have also been investigated as far as regarding the search for life on other planets and even evaluating the hypothesis that life on Earth originally came from space. The first extreme environments to be largely investigated were those characterized by elevated temperatures. The naturally “hot environments” on Earth range from solar heated surface soils and water with temperatures up to 65 °C, subterranean sites such as oil reserves and terrestrial geothermal with temperatures ranging from slightly above ambient to above 100 °C, to submarine hydrothermal systems with temperatures exceeding 300 °C. There are also human-made environments with elevated temperatures such as compost piles, slag heaps, industrial processes and water heaters. Thermophilic anaerobic microorganisms have been known for a long time, but scientists have often resisted the belief that some organisms do not only survive at high temperatures, but actually thrive under those hot conditions. They are perhaps one of the most interesting varieties of extremophilic organisms. These microorganisms can thrive at temperatures over 50 °C and, based on their optimal temperature, anaerobic thermophiles can be subdivided into three main groups: thermophiles with an optimal temperature between 50 °C and 64 °C and a maximum at 70 °C, extreme thermophiles with an optimal temperature between 65 °C and 80 °C, and finally hyperthermophiles with an optimal temperature above 80 °C and a maximum above 90 °C. The finding of novel extremely thermophilic and hyperthermophilic anaerobic bacteria in recent years, and the fact that a large fraction of them belong

Aerobic Oxidation of Xylose to Xylaric acid in Water over Pt Catalysts.

Science.gov (United States)

Saha, Basudeb; Sadula, Sunitha

2018-05-02

Energy-efficient catalytic conversion of biomass intermediates to functional chemicals can enable bio-products viable. Herein, we report an efficient and low temperature aerobic oxidation of xylose to xylaric acid, a promising bio-based chemical for the production of glutaric acid, over commercial catalysts in water. Among several heterogeneous catalysts investigated, Pt/C exhibits the best activity. Systematic variation of reaction parameters in the pH range of 2.5 to 10 suggests that the reaction is fast at higher temperatures but high C-C scission of intermediate C5-oxidized products to low carbon carboxylic acids undermines xylaric acid selectivity. The C-C cleavage is also high in basic solution. The oxidation at neutral pH and 60 C achieves the highest xylaric acid yield (64%). O2 pressure and Pt-amount have significant influence on the reactivity. Decarboxylation of short chain carboxylic acids results in formation of CO2, causing some carbon loss; however such decarboxylation is slow in the presence of xylose. The catalyst retained comparable activity, in terms of product selectivity, after five cycles with no sign of Pt leaching. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Diversity of Cultured Thermophilic Anaerobes in Hot Springs of Yunnan Province, China

Science.gov (United States)

Lin, L.; Lu, Y.; Dong, X.; Liu, X.; Wei, Y.; Ji, X.; Zhang, C.

2010-12-01

Thermophilic anaerobes including Archaea and Bacteria refer to those growing optimally at temperatures above 50°C and do not use oxygen as the terminal electron acceptor for growth. Study on thermophilic anaerobes will help to understand how life thrives under extreme conditions. Meanwhile thermophilic anaerobes are of importance in potential application and development of thermophilic biotechnology. We have surveyed culturable thermophilic anaerobes in hot springs (pH6.5-7.5; 70 - 94°C) in Rehai of Tengchong, Bangnazhang of Longlin, Eryuan of Dali,Yunnan, China. 50 strains in total were cultured from the hot springs water using Hungate anaerobic technique, and 30 strains were selected based on phenotypic diversity for analysis of 16S rDNA sequences. Phylogenetic analysis showed that 28 strains belonged to the members of five genera: Caldanaerobacter, Calaramator, Thermoanaerobacter, Dictyoglomus and Fervidobacterium, which formed five branches on the phylogenetic tree. Besides, 2 strains of methanogenic archaea were obtained. The majority of the isolates were the known species, however, seven strains were identified as novel species affiliated to the five genera based on the lower 16S rDNA sequence similarities (less than 93 - 97%) with the described species. This work would provide the future study on their diversity, distribution among different regions and the potential application of thermophilic enzyme. Supported by State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences(SKLMR-080605)and the Foundation of State Natural Science (30660009, 30960022, 31081220175).

Clostridium scatologenes strain SL1 isolated as an acetogenic bacterium from acidic sediments.

Science.gov (United States)

Küsel, K; Dorsch, T; Acker, G; Stackebrandt, E; Drake, H L

2000-03-01

A strictly anaerobic, H2-utilizing bacterium, strain SL1, was isolated from the sediment of an acidic coal mine pond. Cells of strain SL1 were sporulating, motile, long rods with a multilayer cell wall. Growth was observed at 5-35 degrees C and pH 3.9-7.0. Acetate was the sole end product of H2 utilization and was produced in stoichiometries indicative of an acetyl-CoA-pathway-dependent metabolism. Growth and substrate utilization also occurred with CO/CO2, vanillate, syringate, ferulate, ethanol, propanol, 1-butanol, glycerine, cellobiose, glucose, fructose, mannose, xylose, formate, lactate, pyruvate and gluconate. With most substrates, acetate was the main or sole product formed. Growth in the presence of H2/CO2 or CO/CO2 was difficult to maintain in laboratory cultures. Methoxyl, carboxyl and acrylate groups of various aromatic compounds were O-demethylated, decarboxylated and reduced, respectively. Small amounts of butyrate were produced during the fermentation of sugars. The acrylate group of ferulate was reduced. Nitrate, sulfate, thiosulfate, dimethylsulfoxide and Fe(III) were not utilized as electron acceptors. Analysis of the 16S rRNA gene sequence of strain SL1 demonstrated that it is closely related to Clostridium scatologenes (99.6% sequence similarity), an organism characterized as a fermentative anaerobe but not previously shown to be capable of acetogenic growth. Comparative experiments with C. scatologenes DSM 757T demonstrated that it utilized H2/CO2 (negligible growth), CO/CO2 (negligible growth), formate, ethanol and aromatic compounds according to stoichiometries indicative of the acetyl-CoA pathway. CO dehydrogenase, formate dehydrogenase and hydrogenase activities were present in both strain SL1 and C. scatologenes DSM 757T. These results indicate that (i) sediments of acidic coal mine ponds harbour acetogens and (ii) C. scatologenes is an acetogen that tends to lose its capacity to grow acetogenically under H2/CO2 or CO/CO2 after prolonged

[Current clinical significance of anaerobic bacteremia].

Science.gov (United States)

Jirsa, Roman; Marešová, Veronika; Brož, Zdeněk

2010-10-01

to estimate tje current clinical significance of anaerobic bacteremia in a group of Czech hospitals. this retrospective analysis comprised 8 444 anaerobic blood cultures in patients admitted to four Czech hospitals between 2004 and 2007. in 16 patients, blood cultures yielded significant anaerobic bacteria. Thus, anaerobic bacteremia accounted for less than 2 % of clinically significant bacteremia. Four patients (18 %) died but none of the deaths could be clearly attributable to anaerobic bacteria in the bloodstream. The most common comorbidities predisposing to anaerobic bacteremia and the most frequent sources of infection were similar to those reported by other authors. The majority of anaerobic bacteremia cases were due to gram-negative bacteria, followed by Clostridium perfringens and, surprisingly, Eubacterium spp. (particularly Eubacterium lentum). anaerobic bacteremia remains rare. The comparison of our data with those by other authors suggests that (despite the reported high mortality) the actual clinical significance of anaerobic bacteremia is rather controversial and that the anaerobic bacteremia might not correspond to more serious pathogenic role of the anaerobic bacteria as the source of infection.

Single-cell analysis of growth and cell division of the anaerobe Desulfovibrio vulgaris Hildenborough

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Anouchka eFievet

2015-12-01

Full Text Available Recent years have seen significant progress in understanding basic bacterial cell cycle properties such as cell growth and cell division. While characterization and regulation of bacterial cell cycle is quite well documented in the case of fast growing aerobic model organisms, no data has been so far reported for anaerobic bacteria. This lack of information in anaerobic microorganisms can mainly be explained by the absence of molecular and cellular tools such as single cell microscopy and fluorescent probes usable for anaerobes and essential to study cellular events and/or subcellular localization of the actors involved in cell cycle.In this study, single-cell microscopy has been adapted to study for the first time, in real time, the cell cycle of a bacterial anaerobe, Desulfovibrio vulgaris Hildenborough (DvH. This single-cell analysis provides mechanistic insights into the cell division cycle of DvH, which seems to be governed by the recently discussed so-called incremental model that generates remarkably homogeneous cell sizes. Furthermore, cell division was reversibly blocked during oxygen exposure. This may constitute a strategy for anaerobic cells to cope with transient exposure to oxygen that they may encounter in their natural environment, thereby contributing to their aerotolerance. This study lays the foundation for the first molecular, single-cell assay that will address factors that cannot otherwise be resolved in bulk assays and that will allow visualization of a wide range of molecular mechanisms within living anaerobic cells.

Urea utilization in growing lambs. 6

International Nuclear Information System (INIS)

Ulbrich, M.; Nikitin, S.; Geissler, C.; Hoffmann, M.

1988-01-01

Lambs aged 2 or 4 months and of an average live weight of 14.7 and 27.4 kg, resp., received rations consisting of 44% cereals, 46% dried sugar beet pulp, 6% wheat starch, 2% urea and 2% mineral-vitamin mixture. The crude protein content was 17.1 and 15.9%, resp., in the dry matter, that of natively crude protein 10.6 and 9.4%, resp. During a 6-day N balance period 8 and 16 g 15 N-urea resp. with a 15 N excess ( 15 N') of 9.26 and 9.40 atom-% were fed orally instead of commercial feeding urea. There were no significant differences between the two age groups with regard to the digestibility of the organic matter and the crude nutrients. The average N balance of 372 ± 85 mg/kg LW 0.75 /day were in the intermediate range of N retention capacity and accounted for 26 ± 5% of the consumed N. N retention in per cent. was slightly lower in younger lambs. Projections of urea utilization in a quasi stationary state resulted in an efficiency of the utilization of 33 ± 4%. The cutting up of the lambs at the end of the main period showed between 0.02 and 0.22 atom-% 15 N' in the total N, TCA precipitable N and amino acid N of the meat. At between 0.24 and 0.38 atom-% 15 N' they were highest in the heart and jaw muscles. The quota of 15 N' amounts found in the total N of the meat were 10.6 ± 3% of the 15 N intake and 20.1 ± 5.1% of the 15 N'amount remaining in the body. The bones contained 7.7 ± 1.7% and the fleece 7.9 ± 3.1% of the 15 N' intake. Total N and urea utilization was slightly lower in younger lambs than in older ones. (author)

Bio-recovery of N and P from an anaerobic digester effluent: The ...

African Journals Online (AJOL)

The results obtained indicated that duckweed was capable of growing on the anaerobic digester effluent provided its TAN content did not exceed 42 mg N l-1. Nitrogen uptake by the duckweed from the effluent ranged between 53 and 115.7 mg l-1 whereas P uptake varied from 1.40 to 8.4 mg P l-1. The relative growth rate ...

Anaerobe Tolerance to Oxygen and the Potentials of Anaerobic and Aerobic Cocultures for Wastewater Treatment

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M.T. Kato

1997-12-01

Full Text Available The anaerobic treatment processes are considered to be well-established methods for the elimination of easily biodegradable organic matter from wastewaters. Some difficulties concerning certain wastewaters are related to the possible presence of dissolved oxygen. The common belief is that anaerobes are oxygen intolerant. Therefore, the common practice is to use sequencing anaerobic and aerobic steps in separate tanks. Enhanced treatment by polishing off the residual biodegradable oxygen demand from effluents of anaerobic reactors, or the biodegradation of recalcitrant wastewater pollutants, usually requires sequenced anaerobic and aerobic bacteria activities. However, the combined activity of both bacteria can also be obtained in a single reactor. Previous experiments with either pure or mixed cultures showed that anaerobes can tolerate oxygen to a certain extent. The oxygen toxicity to methanogens in anaerobic sludges was quantified in batch experiments, as well as in anaerobic reactors. The results showed that methanogens have a high tolerance to oxygen. In practice, it was confirmed that dissolved oxygen does not constitute any detrimental effect on reactor treatment performance. This means that the coexistence of anaerobic and aerobic bacteria in one single reactor is feasible and increases the potentials of new applications in wastewater treatment

Anaerobic fungal populations

International Nuclear Information System (INIS)

Brookman, J.L.; Nicholson, M.J.

2005-01-01

The development of molecular techniques has greatly broadened our view of microbial diversity and enabled a more complete detection and description of microbial communities. The application of these techniques provides a simple means of following community changes, for example, Ishii et al. described transient and more stable inhabitants in another dynamic microbial system, compost. Our present knowledge of anaerobic gut fungal population diversity within the gastrointestinal tract is based upon isolation, cultivation and observations in vivo. It is likely that there are many species yet to be described, some of which may be non-culturable. We have observed a distinct difference in the ease of cultivation between the different genera, for example, Caecomyes isolates are especially difficult to isolate and maintain in vitro, a feature that is likely to result in the under representation of this genera in culture-based enumerations. The anaerobic gut fungi are the only known obligately anaerobic fungi. For the majority of their life cycles, they are found tightly associated with solid digesta in the rumen and/or hindgut. They produce potent fibrolytic enzymes and grow invasively on and into the plant material they are digesting making them important contributors to fibre digestion. This close association with intestinal digesta has made it difficult to accurately determine the amount of fungal biomass present in the rumen, with Orpin suggesting 8% contribution to the total microbial biomass, whereas Rezaeian et al. more recently gave a value of approximately 20%. It is clear that the rumen microbial complement is affected by dietary changes, and that the fungi are more important in digestion in the rumens of animals fed with high-fibre diets. It seems likely that the gut fungi play an important role within the rumen as primary colonizers of plant fibre, and so we are particularly interested in being able to measure the appearance and diversity of fungi on the plant

Enzymatic Xylose Release from Pretreated Corn Bran Arabinoxylan: Differential Effects of Deacetylation and Deferuloylation on Insoluble and Soluble Substrate Fractions

DEFF Research Database (Denmark)

Agger, Jane; Viksø-Nielsen, Ander; Meyer, Anne S.

2010-01-01

In the present work enzymatic hydrolysis of arabinoxylan from pretreated corn bran (190 °C, 10 min) was evaluated by measuring the release of xylose and arabinose after treatment with a designed minimal mixture of monocomponent enzymes consisting of α-l-arabinofuranosidases, an endoxylanase......, and a β-xylosidase. The pretreatment divided the corn bran material 50:50 into soluble and insoluble fractions having A:X ratios of 0.66 and 0.40, respectively. Addition of acetyl xylan esterase to the monocomponent enzyme mixture almost doubled the xylose release from the insoluble substrate fraction...

Dual utilization of NADPH and NADH cofactors enhances xylitol production in engineered Saccharomyces cerevisiae.

Science.gov (United States)

Jo, Jung-Hyun; Oh, Sun-Young; Lee, Hyeun-Soo; Park, Yong-Cheol; Seo, Jin-Ho

2015-12-01

Xylitol, a natural sweetener, can be produced by hydrogenation of xylose in hemicelluloses. In microbial processes, utilization of only NADPH cofactor limited commercialization of xylitol biosynthesis. To overcome this drawback, Saccharomyces cerevisiae D452-2 was engineered to express two types of xylose reductase (XR) with either NADPH-dependence or NADH-preference. Engineered S. cerevisiae DWM expressing both the XRs exhibited higher xylitol productivity than the yeast strain expressing NADPH-dependent XR only (DWW) in both batch and glucose-limited fed-batch cultures. Furthermore, the coexpression of S. cerevisiae ZWF1 and ACS1 genes in the DWM strain increased intracellular concentrations of NADPH and NADH and improved maximum xylitol productivity by 17%, relative to that for the DWM strain. Finally, the optimized fed-batch fermentation of S. cerevisiae DWM-ZWF1-ACS1 resulted in 196.2 g/L xylitol concentration, 4.27 g/L h productivity and almost the theoretical yield. Expression of the two types of XR utilizing both NADPH and NADH is a promising strategy to meet the industrial demands for microbial xylitol production. Copyright © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Utilization of Urea Treated and Untreated Cocoa Pod Husk Based Diets by Growing Pigs : An On-farm Study

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Iyayi, EA.

2001-01-01

Full Text Available An on-farm adaptation study of the utilization of urea treated and untreated cocoa pod husk (CPH by growing pigs was carried out on a commercial pig farm. Thirty-two Landrace X Large White growing pigs (16 males + 16 females were randomly assigned to 4 experimental diets. Diet 1 was a standard grower ration (control. In Diets 2 and 3 CPH meal was included at 250 g/kg, that used in Diet 3 being treated with a 5 % urea solution. Diet 4 was the farmer's diet. There was no significant difference (P> 0.05 between diets 3 and the control in their effect on the performance of the animals. These two diets caused a better (P 0.05 influence on the backfat thickness. Carcass cuts were also not significantly (P> 0.05 influenced by inclusion of CPH meal. Results suggest (1 the possibility of formulating diets for growing pigs using CPH meals and (2 that further treatment of the CPH meal with urea improves its nutritive value resulting in better performance and economy of production.

Strain-resolved microbial community proteomics reveals simultaneous aerobic and anaerobic function during gastrointestinal tract colonization of a preterm infant

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Brandon eBrooks

2015-07-01

Full Text Available While there has been growing interest in the gut microbiome in recent years, it remains unclear whether closely related species and strains have similar or distinct functional roles and if organisms capable of both aerobic and anaerobic growth do so simultaneously. To investigate these questions, we implemented a high-throughput mass spectrometry-based proteomics approach to identify proteins in fecal samples collected on days of life 13-21 from an infant born at 28 weeks gestation. No prior studies have coupled strain-resolved community metagenomics to proteomics for such a purpose. Sequences were manually curated to resolve the genomes of two strains of Citrobacter that were present during the later stage of colonization. Proteome extracts from fecal samples were processed via a nano-2D-LC-MS/MS and peptides were identified based on information predicted from the genome sequences for the dominant organisms, Serratia and the two Citrobacter strains. These organisms are facultative anaerobes, and proteomic information indicates the utilization of both aerobic and anaerobic metabolisms throughout the time series. This may indicate growth in distinct niches within the gastrointestinal tract. We uncovered differences in the physiology of coexisting Citrobacter strains, including differences in motility and chemotaxis functions. Additionally, for both Citrobacter strains we resolved a community-essential role in vitamin metabolism and a predominant role in propionate production. Finally, in this case study we detected differences between genome abundance and activity levels for the dominant populations. This underlines the value in layering proteomic information over genetic potential.

Effect of high-voltage pulsed electric field (HPEF pretreatment on biogas production rates of hybrid Pennisetum by anaerobic fermentation

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Baijuan Wang

2018-02-01

Full Text Available In this paper, the raw materials of hybrid Pennisetum were pretreated in different conditions of high voltage pulsed electric field (HPEF to improve its material utilization ratios and biogas production rates of anaerobic fermentation. Then, anaerobic digestion experiments were conducted within 32 days at moderate temperature (35 °C with TS mass fraction (6%, inoculation rate (20% and initial pH (7.0. It is indicated that compared with the control group, 9 groups of hybrid Pennisetum pretreated by HPEF are obviously superior in gas production efficiency of anaerobic fermentation, and higher in cumulative gas production, peak daily gas production and maximum methane concentration; that the most remarkable stimulation occurs in the HPEF condition of 15 kV/120 Hz/60 min, in that situation, the cumulative gas production in the fermentation period of 32 days is up to 9587 mL, 26.95% higher than that of the control group, the peak daily gas production increases and the range of peak period extends. It is demonstrated that the optimal HPEF pretreatment time is 60 min and three HPEF parameters have a better effect on gas production in the order of voltage > time > frequency; and that the effect degree of treatment parameters on peak daily gas production is voltage, frequency and time in turn. It is concluded that HPEF can improve material utilization ratio and gas production rate of hybrid Pennisetum by anaerobic fermentation and shorten the gas production cycle. By virtue of this physical pretreatment method, the resource of Pennisetum is utilized sufficiently and the classes of energy plants are enlarged effectively. Keywords: Hybrid Pennisetum, Anaerobic fermentation, High voltage pulsed electric field (HPEF, Biogas, Material utilization ratio, Gas generation rate, Model, Stimulation

Anaerobic treatment techniques

International Nuclear Information System (INIS)

Boehnke, B.; Bischofsberger, W.; Seyfried, C.F.

1993-01-01

This practical and theoretical guide presents the current state of knowledge in anaerobic treatment of industrial effluents with a high organic pollutant load and sewage sludges resulting from the treatment of municipal and industrial waste water. Starting from the microbiological bases of anaerobic degradation processes including a description and critical evaluation of executed plants, the book evolves the process-technical bases of anaerobic treatment techniques, derives relative applications, and discusses these with reference to excuted examples. (orig./UWA). 232 figs [de

Combination of anaerobic effluent and lignocellulosic bacterial consortium to reduce vermicomposting time

Science.gov (United States)

Utilization of solid bio-fertilizers is an alternative to avoid chemical degradation of soil. Anaerobic biodigestor effluents/digestates have been used effectively as fertilizers. However, they may have several risk factors such as the presence of pathogens and heavy metals. Vermicomposting could he...

Comparative economic analysis: Anaerobic digester case study

International Nuclear Information System (INIS)

Lusk, P.D.

1991-01-01

An economic guide is developed to assess the value of anaerobic digesters used on dairy farms. Two varieties of anaerobic digesters, a conventional mixed-tank mesophilic and an innovative earthen psychrophilic, are comparatively evaluated using a cost-effectiveness index. The two case study examples are also evaluated using three other investment merit statistics: simple payback period, net present value, and internal rate of return. Life-cycle savings are estimated for both varieties, with sensitivities considered for investment risk. The conclusion is that an earthen psychrophilic digester can have a significant economic advantage over a mixed-tank mesophilic digester because of lower capital cost and reduced operation and maintenance expenses. Because of this economic advantage, additional projects are being conducted in North Carolina to increase the rate of biogas utilization. The initial step includes using biogas for milk cooling at the dairy farm where the existing psychrophilic digester is located. Further, a new project is being initiated for electricity production with thermal reclaim at a swine operation

Furfural synthesis from D-xylose in the presence of sodium chloride : Microwave versus conventional heating

NARCIS (Netherlands)

Xiouras, C.; Radacsi, N.; Sturm, G.S.J.; Stefanidis, G.

2016-01-01

We investigate the existence of specific/nonthermal microwave effects for the dehydration reaction of xylose to furfural in the presence of NaCl. Such effects are reported for sugars dehydration reactions in several literature reports. To this end, we adopted three approaches that compare

Shotgun Proteomics of Aspergillus niger Microsomes upon d-Xylose Induction▿ †

Science.gov (United States)

de Oliveira, José Miguel P. Ferreira; van Passel, Mark W. J.; Schaap, Peter J.; de Graaff, Leo H.

2010-01-01

Protein secretion plays an eminent role in cell maintenance and adaptation to the extracellular environment of microorganisms. Although protein secretion is an extremely efficient process in filamentous fungi, the mechanisms underlying protein secretion have remained largely uncharacterized in these organisms. In this study, we analyzed the effects of the d-xylose induction of cellulase and hemicellulase enzyme secretion on the protein composition of secretory organelles in Aspergillus niger. We aimed to systematically identify the components involved in the secretion of these enzymes via mass spectrometry of enriched subcellular microsomal fractions. Under each condition, fractions enriched for secretory organelles were processed for tandem mass spectrometry, resulting in the identification of peptides that originate from 1,081 proteins, 254 of which—many of them hypothetical proteins—were predicted to play direct roles in the secretory pathway. d-Xylose induction led to an increase in specific small GTPases known to be associated with polarized growth, exocytosis, and endocytosis. Moreover, the endoplasmic-reticulum-associated degradation (ERAD) components Cdc48 and all 14 of the 20S proteasomal subunits were recruited to the secretory organelles. In conclusion, induction of extracellular enzymes results in specific changes in the secretory subproteome of A. niger, and the most prominent change found in this study was the recruitment of the 20S proteasomal subunits to the secretory organelles. PMID:20453123

The Effect of Anaerobic and Aerobic Fish Sludge Supernatant on Hydroponic Lettuce

NARCIS (Netherlands)

Goddek, Simon; Schmautz, Zala; Scott, Ben; Delaide, Boris; Keesman, Karel; Wuertz, Sven; Junge, Ranka

2016-01-01

The mobilization of nutrients from fish sludge (i.e., feces and uneaten feed) plays a key role in optimizing the resource utilization and thus in improving the sustainability of aquaponic systems. While several studies have documented the aerobic and anaerobic digestion performance of aquaculture

Sequential ethanol fermentation and anaerobic digestion increases bioenergy yields from duckweed.

Science.gov (United States)

Calicioglu, O; Brennan, R A

2018-06-01

The potential for improving bioenergy yields from duckweed, a fast-growing, simple, floating aquatic plant, was evaluated by subjecting the dried biomass directly to anaerobic digestion, or sequentially to ethanol fermentation and then anaerobic digestion, after evaporating ethanol from the fermentation broth. Bioethanol yields of 0.41 ± 0.03 g/g and 0.50 ± 0.01 g/g (glucose) were achieved for duckweed harvested from the Penn State Living-Filter (Lemna obscura) and Eco-Machine™ (Lemna minor/japonica and Wolffia columbiana), respectively. The highest biomethane yield, 390 ± 0.1 ml CH 4 /g volatile solids added, was achieved in a reactor containing fermented duckweed from the Living-Filter at a substrate-to-inoculum (S/I) ratio (i.e., duckweed to microorganism ratio) of 1.0. This value was 51.2% higher than the biomethane yield of a replicate reactor with raw (non-fermented) duckweed. The combined bioethanol-biomethane process yielded 70.4% more bioenergy from duckweed, than if anaerobic digestion had been run alone. Copyright © 2018 Elsevier Ltd. All rights reserved.

Anaerobic Psychrophiles from Lake Zub and Lake Untersee, Antarctica

Science.gov (United States)

Townsend, Alisa; Pikuta, Elena V.; Guisler, Melissa; Stahl, Sarah; Hoover, Richard B.

2009-01-01

The study of samples from Antarctica 2008 and 2009 expeditions organized and successfully conducted by Richard Hoover led to the isolation of diverse anaerobic strains with psychrotolerant and psychrophilic physiology. Due to the fact that Lake Untersee has never been subject to microbiological study, this work with the samples has significant and pioneering impact to the knowledge about the biology of this unique ecosystem. Also, the astrobiological significance for the study of these ecosystems is based on new findings of ice covered water systems on other bodies of our solar system. Anaerobic psychrotolerant strain LZ-22 was isolated from a frozen sample of green moss with soils around the rhizosphere collected near Lake Zub in Antarctica. Morphology of strain LZ-22 was observed to be motile, rod shaped and spore-forming cells with sizes 1 x 5-10 micron. This new isolate is a mesophile with the maximum temperature of growth at 40C. Strain LZ-22 is able to live on media without NaCl and in media with up to 7% (w/v) NaCl. It is catalase negative and grows only on sugars with the best growth rate being on lactose. The strain is a neutrophile and grows between pH 5 and 9.0 with the optimum at 7.8. Another two strains UL7-96mG and LU-96m7P were isolated from deep water samples of Lake Untersee. Proteolytic strain LU-96m7P had a truly psychrophilic nature and refused to grow at room temperature. Sugarlytic strain UL7-96mG was found to be psychrotolerant, but its rate of growth at 3C was very high compared with other mesophiles. Two homoacetogenic psychrophilic strains A7AC-96m and AC-DS7 were isolated and purified from samples of Lake Untersee; both of them are able to grow chemolithotrophically on H2+CO2. In the presence of lactate, these strains are able to grow only at 0-18C, and growth at 22C was observed only with yeast extract stimulation. In this paper, physiological and morphological characteristics of novel psychrophilic and psychrotolerant isolates from

Ethanol production from wet-exploded wheat straw hydrolysate by thermophilic anaerobic bacterium Thermoanaerobacter BG1L1 in a continuous immobilized reactor

DEFF Research Database (Denmark)

Georgieva, Tania I.; Mikkelsen, Marie Just; Ahring, Birgitte Kiær

2008-01-01

was not detoxified, ethanol yield in a range of 0.39-0.42 g/g was obtained. Overall, sugar efficiency to ethanol was 68-76%. The reactor was operated continuously for approximately 143 days, and no contamination was seen without the use of any agent for preventing bacterial infections. The tested microorganism has......Thermophilic ethanol fermentation of wet-exploded wheat straw hydrolysate was investigated in a continuous immobilized reactor system. The experiments were carried out in a lab-scale fluidized bed reactor (FBR) at 70C. Undetoxified wheat straw hydrolysate was used (3-12% dry matter), corresponding...... to sugar mixtures of glucose and xylose ranging from 12 to 41 g/l. The organism, thermophilic anaerobic bacterium Thermoanaerobacter BG1L1, exhibited significant resistance to high levels of acetic acid (up to 10 g/l) and other metabolic inhibitors present in the hydrolysate. Although the hydrolysate...

Clinical features of anaerobic orthopaedic infections.

Science.gov (United States)

Lebowitz, Dan; Kressmann, Benjamin; Gjoni, Shpresa; Zenelaj, Besa; Grosgurin, Olivier; Marti, Christophe; Zingg, Matthieu; Uçkay, Ilker

2017-02-01

Some patient populations and types of orthopaedic surgery could be at particular risk for anaerobic infections. In this retrospective cohort study of operated adult patients with infections from 2004 to 2014, we assessed obligate anaerobes and considered first clinical infection episodes. Anaerobes, isolated from intra-operative samples, were identified in 2.4% of 2740 surgical procedures, of which half (33/65; 51%) were anaerobic monomicrobial infections. Propionibacterium acnes, a penicillin and vancomycin susceptible pathogen, was the predominantly isolated anaerobe. By multivariate analysis, the presence of fracture fixation plates was the variable most strongly associated with anaerobic infection (odds ratio: 2.1, 95% CI: 1.3-3.5). Anaerobes were also associated with spondylodesis and polymicrobial infections. In contrast, it revealed less likely in native bone or prosthetic joint infections and was not related to prior antibiotic use. In conclusion, obligate anaerobes in our case series of orthopaedic infections were rare, and mostly encountered in infections related to trauma with open-fracture fixation devices rather than clean surgical site infection. Anaerobes were often co-pathogens, and cultures most frequently recovered P. acnes. These observations thus do not support changes in current practices such as broader anaerobe coverage for perioperative prophylaxis.

RISK FACTORS IN NEONATAL ANAEROBIC INFECTIONS

Directory of Open Access Journals (Sweden)

M. S. Tabib

2008-06-01

Full Text Available Anaerobic bacteria are well known causes of sepsis in adults but there are few studies regarding their role in neonatal sepsis. In an attempt to define the incidence of neonatal anaerobic infections a prospective study was performed during one year period. A total number of 400 neonates under sepsis study were entered this investigation. Anaerobic as well as aerobic cultures were sent. The patients were subjected to comparison in two groups: anaerobic culture positive and anaerobic culture negative and this comparison were analyzed statistically. There were 7 neonates with positive anaerobic culture and 35 neonates with positive aerobic culture. A significant statistical relationship was found between anaerobic infections and abdominal distention and pneumonia. It is recommended for those neonates with abdominal distention and pneumonia refractory to antibiotic treatment to be started on antibiotics with anaerobic coverage.

Evaluation of a kinetic model for computer simulation of growth and fermentation by Scheffersomyces (Pichia) stipitis fed D-xylose.

Science.gov (United States)

Slininger, P J; Dien, B S; Lomont, J M; Bothast, R J; Ladisch, M R; Okos, M R

2014-08-01

Scheffersomyces (formerly Pichia) stipitis is a potential biocatalyst for converting lignocelluloses to ethanol because the yeast natively ferments xylose. An unstructured kinetic model based upon a system of linear differential equations has been formulated that describes growth and ethanol production as functions of ethanol, oxygen, and xylose concentrations for both growth and fermentation stages. The model was validated for various growth conditions including batch, cell recycle, batch with in situ ethanol removal and fed-batch. The model provides a summary of basic physiological yeast properties and is an important tool for simulating and optimizing various culture conditions and evaluating various bioreactor designs for ethanol production. © 2014 Wiley Periodicals, Inc.

Microbial ecology of anaerobic digesters: the key players of anaerobiosis.

Science.gov (United States)

Ali Shah, Fayyaz; Mahmood, Qaisar; Maroof Shah, Mohammad; Pervez, Arshid; Ahmad Asad, Saeed

2014-01-01

Anaerobic digestion is the method of wastes treatment aimed at a reduction of their hazardous effects on the biosphere. The mutualistic behavior of various anaerobic microorganisms results in the decomposition of complex organic substances into simple, chemically stabilized compounds, mainly methane and CO2. The conversions of complex organic compounds to CH4 and CO2 are possible due to the cooperation of four different groups of microorganisms, that is, fermentative, syntrophic, acetogenic, and methanogenic bacteria. Microbes adopt various pathways to evade from the unfavorable conditions in the anaerobic digester like competition between sulfate reducing bacteria (SRB) and methane forming bacteria for the same substrate. Methanosarcina are able to use both acetoclastic and hydrogenotrophic pathways for methane production. This review highlights the cellulosic microorganisms, structure of cellulose, inoculum to substrate ratio, and source of inoculum and its effect on methanogenesis. The molecular techniques such as DGGE (denaturing gradient gel electrophoresis) utilized for dynamic changes in microbial communities and FISH (fluorescent in situ hybridization) that deal with taxonomy and interaction and distribution of tropic groups used are also discussed.

Microbial Ecology of Anaerobic Digesters: The Key Players of Anaerobiosis

Science.gov (United States)

Ali Shah, Fayyaz; Mahmood, Qaisar; Maroof Shah, Mohammad; Pervez, Arshid; Ahmad Asad, Saeed

2014-01-01

Anaerobic digestion is the method of wastes treatment aimed at a reduction of their hazardous effects on the biosphere. The mutualistic behavior of various anaerobic microorganisms results in the decomposition of complex organic substances into simple, chemically stabilized compounds, mainly methane and CO2. The conversions of complex organic compounds to CH4 and CO2 are possible due to the cooperation of four different groups of microorganisms, that is, fermentative, syntrophic, acetogenic, and methanogenic bacteria. Microbes adopt various pathways to evade from the unfavorable conditions in the anaerobic digester like competition between sulfate reducing bacteria (SRB) and methane forming bacteria for the same substrate. Methanosarcina are able to use both acetoclastic and hydrogenotrophic pathways for methane production. This review highlights the cellulosic microorganisms, structure of cellulose, inoculum to substrate ratio, and source of inoculum and its effect on methanogenesis. The molecular techniques such as DGGE (denaturing gradient gel electrophoresis) utilized for dynamic changes in microbial communities and FISH (fluorescent in situ hybridization) that deal with taxonomy and interaction and distribution of tropic groups used are also discussed. PMID:24701142

Tolerance of anaerobic bacteria to chlorinated solvents.

Science.gov (United States)

Koenig, Joanna C; Groissmeier, Kathrin D; Manefield, Mike J

2014-01-01

The aim of this research was to evaluate the effects of four chlorinated aliphatic hydrocarbons (CAHs), perchloroethene (PCE), carbon tetrachloride (CT), chloroform (CF) and 1,2-dichloroethane (1,2-DCA), on the growth of eight anaerobic bacteria: four fermentative species (Escherichia coli, Klebsiella sp., Clostridium sp. and Paenibacillus sp.) and four respiring species (Pseudomonas aeruginosa, Geobacter sulfurreducens, Shewanella oneidensis and Desulfovibrio vulgaris). Effective concentrations of solvents which inhibited growth rates by 50% (EC50) were determined. The octanol-water partition coefficient or log Po/w of a CAH proved a generally satisfactory measure of its toxicity. Most species tolerated approximately 3-fold and 10-fold higher concentrations of the two relatively more polar CAHs CF and 1,2-DCA, respectively, than the two relatively less polar compounds PCE and CT. EC50 values correlated well with growth rates observed in solvent-free cultures, with fast-growing organisms displaying higher tolerance levels. Overall, fermentative bacteria were more tolerant to CAHs than respiring species, with iron- and sulfate-reducing bacteria in particular appearing highly sensitive to CAHs. These data extend the current understanding of the impact of CAHs on a range of anaerobic bacteria, which will benefit the field of bioremediation.

Kinetic modeling of the effect of solids retention time on methanethiol dynamics in anaerobic digestion.

Science.gov (United States)

Zhang, Dian; Strawn, Mary; Novak, John T; Wang, Zhi-Wu

2018-07-01

The highly volatile methanethiol (MT) with an extremely low odor threshold and distinctive putrid smell is often identified as a major odorous compound generated under anaerobic conditions. As an intermediate compound in the course of anaerobic digestion, the extent of MT emission is closely related to the time of anaerobic reaction. In this study, lab-scale anaerobic digesters were operated at solids retention time (SRTs) of 15, 20, 25, 30, 40 and 50 days to investigate the effect of SRT on MT emission. The experimental results demonstrated a bell-shaped curve of MT emission versus SRT with a peak around 20 days SRT. In order to understand this SRT effect, a kinetic model was developed to describe MT production and utilization dynamics in the course of anaerobic digestion and calibrated with the experimental results collected from this study. The model outcome revealed that the high protein content in the feed sludge together with the large maintenance coefficient of MT fermenters are responsible for the peak MT emission emergence in the range of typical SRT used for anaerobic digestion. A further analysis of the kinetic model shows that it can be extensively simplified with reasonable approximation to a form that anaerobic digestion practitioners could easily use to predict the MT and SRT relationship. Copyright © 2018 Elsevier Ltd. All rights reserved.

Pathways and bioenergetics of anaerobic carbon monoxide fermentation.

OpenAIRE

Martijn eDiender; Alfons J.M. Stams; Alfons J.M. Stams; Diana Z. Sousa

2015-01-01

Carbon monoxide can act as a substrate for different modes of fermentative anaerobic metabolism. The trait of utilizing CO is spread among a diverse group of microorganisms, including members of bacteria as well as archaea. Over the last decade this metabolism has gained interest due to the potential of converting CO rich gas, such as synthesis gas, into bio-based products. Three main types of fermentative CO metabolism can be distinguished: hydrogenogenesis, methanogenesis and acetogenesis, ...

D-Xylose fermentation, xylitol production and xylanase activities by seven new species of Sugiyamaella.

Science.gov (United States)

Sena, Letícia M F; Morais, Camila G; Lopes, Mariana R; Santos, Renata O; Uetanabaro, Ana P T; Morais, Paula B; Vital, Marcos J S; de Morais, Marcos A; Lachance, Marc-André; Rosa, Carlos A

2017-01-01

Sixteen yeast isolates identified as belonging to the genus Sugiyamaella were studied in relation to D-xylose fermentation, xylitol production, and xylanase activities. The yeasts were recovered from rotting wood and sugarcane bagasse samples in different Brazilian regions. Sequence analyses of the internal transcribed spacer (ITS) region and the D1/D2 domains of large subunit rRNA gene showed that these isolates belong to seven new species. The species are described here as Sugiyamaella ayubii f.a., sp. nov. (UFMG-CM-Y607 T  = CBS 14108 T ), Sugiyamaella bahiana f.a., sp. nov. (UFMG-CM-Y304 T  = CBS 13474 T ), Sugiyamaella bonitensis f.a., sp. nov. (UFMG-CM-Y608 T  = CBS 14270 T ), Sugiyamaella carassensis f.a., sp. nov. (UFMG-CM-Y606 T  = CBS 14107 T ), Sugiyamaella ligni f.a., sp. nov. (UFMG-CM-Y295 T  = CBS 13482 T ), Sugiyamaella valenteae f.a., sp. nov. (UFMG-CM-Y609 T  = CBS 14109 T ) and Sugiyamaella xylolytica f.a., sp. nov. (UFMG-CM-Y348 T  = CBS 13493 T ). Strains of the described species S. boreocaroliniensis, S. lignohabitans, S. novakii and S. xylanicola, isolated from rotting wood of Brazilian ecosystems, were also compared for traits relevant to xylose metabolism. S. valenteae sp. nov., S. xylolytica sp. nov., S. bahiana sp. nov., S. bonitensis sp. nov., S. boreocarolinensis, S. lignohabitans and S. xylanicola were able to ferment D-xylose to ethanol. Xylitol production was observed for all Sugiyamaella species studied, except for S. ayubii sp. nov. All species studied showed xylanolytic activity, with S. xylanicola, S. lignohabitans and S. valenteae sp. nov. having the highest values. Our results suggest these Sugiyamaella species have good potential for biotechnological applications.

Anaerobic carbon monoxide metabolism by Pleomorphomonas carboxyditropha sp. nov., a new mesophilic hydrogenogenic carboxydotroph.

Science.gov (United States)

Esquivel-Elizondo, Sofia; Maldonado, Juan; Krajmalnik-Brown, Rosa

2018-06-01

Carbon monoxide (CO)-metabolism and phenotypic and phylogenetic characterization of a novel anaerobic, mesophilic and hydrogenogenic carboxydotroph are reported. Strain SVCO-16 was isolated from anaerobic sludge and grows autotrophically and mixotrophically with CO. The genes cooS and cooF, coding for a CO dehydrogenase complex, and genes similar to hycE2, encoding a CO-induced hydrogenase, were present in its genome. The isolate produces H2 and CO2 from CO, and acetate and formate from organic substrates. Based on the 16S rRNA sequence, it is an Alphaproteobacterium most closely related to the genus Pleomorphomonas (98.9%-99.2% sequence identity). Comparison with other previously characterized Pleomorphomonas showed that P. diazotrophica and P. oryzae do not metabolize CO, and P. diazotrophica does not grow anaerobically with organic substrates. Average nucleotide identity values between strain SVCO-16 and P. diazotrophica, P. oryzae or P. koreensis were 86.66 ± 0.21%. These values are below the boundary to define species (95%-96%). Digital DNA-DNA hybridization estimates between strain SVCO-16 and reference strains were also below the 70% threshold for species delineation: 29.1%-34.5%. Based on the differences in CO metabolism, genome analyses and cellular fatty acid composition, the isolate should be classified into the genus Pleomorphomonas as a representative of a novel species, Pleomorphomonas carboxyditropha. The type strain of Pleomorphomonas carboxyditropha is SVCO-16T (strain deposit numbers, DSM 106132T and TSD-119T).

Isolation and characterization of a new hydrogen-utilizing bacterium from the rumen.

Science.gov (United States)

Rieu-Lesme, F; Fonty, G; Doré, J

1995-01-01

A new H2/CO2-utilizing acetogenic bacterium was isolated from the rumen of a mature deer. This is the first report of a spore-forming Gram-negative bacterial species from the rumen. The organism was a strictly anaerobic, motile rod and was able to grow autotrophically on hydrogen and carbon dioxide. Acetate was the major product detected. Glucose, fructose and lactate were also fermented heterotrophically. The optimum pH for growth was 7.0-7.5, and the optimum temperature was 37-42 degrees C. Yeast extract was required for growth and rumen fluid was highly stimulatory. The DNA base ratio was 52.9 +/- 0.5 mol% G+C. On the basis of these characteristics and fermentation products, the isolate was considered to be different from acetogenic bacteria described previously.

Bioelectrochemical enhancement of anaerobic methanogenesis for high organic load rate wastewater treatment in a up-flow anaerobic sludge blanket (UASB) reactor.

Science.gov (United States)

Zhao, Zhiqiang; Zhang, Yaobin; Chen, Shuo; Quan, Xie; Yu, Qilin

2014-10-17

A coupling process of anaerobic methanogenesis and electromethanogenesis was proposed to treat high organic load rate (OLR) wastewater. During the start-up stage, acetate removal efficiency of the electric-biological reactor (R1) reached the maximization about 19 percentage points higher than that of the control anaerobic reactor without electrodes (R2), and CH4 production rate of R1 also increased about 24.9% at the same time, while additional electric input was 1/1.17 of the extra obtained energy from methane. Coulombic efficiency and current recorded showed that anodic oxidation contributed a dominant part in degrading acetate when the metabolism of methanogens was low during the start-up stage. Along with prolonging operating time, aceticlastic methanogenesis gradually replaced anodic oxidation to become the main pathway of degrading acetate. When the methanogens were inhibited under the acidic conditions, anodic oxidation began to become the main pathway of acetate decomposition again, which ensured the reactor to maintain a stable performance. FISH analysis confirmed that the electric field imposed could enrich the H2/H(+)-utilizing methanogens around the cathode to help for reducing the acidity. This study demonstrated that an anaerobic digester with a pair of electrodes inserted to form a coupling system could enhance methanogenesis and reduce adverse impacts.

Cellulosic ethanol production via consolidated bioprocessing by a novel thermophilic anaerobic bacterium isolated from a Himalayan hot spring.

Science.gov (United States)

Singh, Nisha; Mathur, Anshu S; Tuli, Deepak K; Gupta, Ravi P; Barrow, Colin J; Puri, Munish

2017-01-01

Cellulose-degrading thermophilic anaerobic bacterium as a suitable host for consolidated bioprocessing (CBP) has been proposed as an economically suited platform for the production of second-generation biofuels. To recognize the overall objective of CBP, fermentation using co-culture of different cellulolytic and sugar-fermenting thermophilic anaerobic bacteria has been widely studied as an approach to achieving improved ethanol production. We assessed monoculture and co-culture fermentation of novel thermophilic anaerobic bacterium for ethanol production from real substrates under controlled conditions. In this study, Clostridium sp. DBT-IOC-C19, a cellulose-degrading thermophilic anaerobic bacterium, was isolated from the cellulolytic enrichment cultures obtained from a Himalayan hot spring. Strain DBT-IOC-C19 exhibited a broad substrate spectrum and presented single-step conversion of various cellulosic and hemicellulosic substrates to ethanol, acetate, and lactate with ethanol being the major fermentation product. Additionally, the effect of varying cellulose concentrations on the fermentation performance of the strain was studied, indicating a maximum cellulose utilization ability of 10 g L -1 cellulose. Avicel degradation kinetics of the strain DBT-IOC-C19 displayed 94.6% degradation at 5 g L -1 and 82.74% degradation at 10 g L -1 avicel concentration within 96 h of fermentation. In a comparative study with Clostridium thermocellum DSM 1313, the ethanol and total product concentrations were higher by the newly isolated strain on pretreated rice straw at an equivalent substrate loading. Three different co-culture combinations were used on various substrates that presented two-fold yield improvement than the monoculture during batch fermentation. This study demonstrated the direct fermentation ability of the novel thermophilic anaerobic bacteria on various cellulosic and hemicellulosic substrates into ethanol without the aid of any exogenous enzymes

Waste biorefineries - integrating anaerobic digestion and microalgae cultivation for bioenergy production.

Science.gov (United States)

Chen, Yi-di; Ho, Shih-Hsin; Nagarajan, Dillirani; Ren, Nan-Qi; Chang, Jo-Shu

2018-04-01

Commercialization of microalgal cultivation has been well realized in recent decades with the use of effective strains that can yield the target products, but it is still challenged by the high costs arising from mass production, harvesting, and further processing. Recently, more interest has been directed towards the utilization of waste resources, such as sludge digestate, to enhance the economic feasibility and sustainability of microalgae production. Anaerobic digestion for waste disposal and phototrophic microalgal cultivation are well-characterized technologies in both fields. However, integration of anaerobic digestion and microalgal cultivation to achieve substantial economic and environmental benefits is extremely limited, and thus deserves more attention and research effort. In particular, combining these two makes possible an ideal 'waste biorefinery' model, as the C/N/P content in the anaerobic digestate can be used to produce microalgal biomass that serves as feedstock for biofuels, while biogas upgrading can simultaneously be performed by phototrophic CO 2 fixation during microalgal growth. This review is thus aimed at elucidating recent advances as well as challenges and future directions with regard to waste biorefineries associated with the integration of anaerobic waste treatment and microalgal cultivation for bioenergy production. Copyright © 2017 Elsevier Ltd. All rights reserved.

Synthesis of furfural from xylose, xylan, and biomass using AlCl3·6H2O in biphasic media via xylose isomerization to xylulose.

Science.gov (United States)

Yang, Yu; Hu, Chang-Wei; Abu-Omar, Mahdi M

2012-02-13

Furfural was prepared in high yields (75 %) from the reaction of xylose in a water-tetrahydrofuran biphasic medium containing AlCl(3)·6H2O and NaCl under microwave heating at 140 °C. The reaction profile revealed the formation of xylulose as an intermediate en route to the dehydration product (furfural). The reaction under these conditions reached completion in 45 min. The aqueous phase containing AlCl(3)·6H(2)O and NaCl could be recycled multiple times (>5) without any loss of activity or selectivity for furfural. Extension of this biphasic reaction system to include xylan as the starting material afforded furfural in 64 % yield. The use of corn stover, pinewood, switchgrass, and poplar gave furfural in 55, 38, 56, and 64 % yield, respectively, at 160 °C. Even though AlCl(3)·6H(2)O did not affect the conversion of crystalline cellulose, moderate yields of the by-product 5-hydroxymethylfurfural (HMF) were noted. The highest HMF yield of 42 % was obtained from pinewood. The coproduction of HMF and furfural from biomass was attributed to the weakening of the cellulose network in the biomass, as a result of hemicellulose hydrolysis. The multifunctional capacity of AlCl(3)·6H(2)O (hemicellulose hydrolysis, xylose isomerization, and xylulose dehydration) in combination with its ease of recyclability make it an attractive candidate/catalyst for the selective synthesis of furfural from various biomass feedstocks. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A growing danger: the risks posed by marihuana grow-ops

Energy Technology Data Exchange (ETDEWEB)

Bradley, F. [Canadian Electricity Association (Canada)

2005-02-01

The proliferation of sophisticated illegal indoor multi-plant marihuana cultivation operations is discussed, focusing primarily on public health and safety issues. Public health issues arise from the high level of molds and pollens caused by high humidity, which can cause asthma, respiratory conditions and allergies, particularly among children, and the likelihood of deadly levels of carbon monoxide build-up resulting from faulty rerouting of the residence's ventilation system. Safety issues discussed are: fires and electrocutions associated with the use of electrical diversions or bipasses to circumvent utility meters, the chemical and electrical hazards involved in investigating and dismantling growing operations, the significant dangers to utility crews who must repair illegal electrical bypasses, injuries by the booby-traps planted to protect the operation from other criminals or law enforcement agents, and the physical danger from the violence, including homicide and assaults, carried out by operators to exert control over production and distribution. Although in general, there is a relaxed attitude towards marihuana use in Canada. there is growing evidence of increasing public concern over large-scale growing operations. Nevertheless, to date operators of grow-ops have been dealt with lightly by the justice system. For example, in British Columbia 11,733 cases have come to the attention of police during the 1997 to 2000 period. Of these about half were dealt with informally (i.e. 'no case' seizures) and 2,255 cases led to at least one offender being convicted. The majority of convictions did not result in custodial dispositions. Only 18 per cent of the cases resulted in prison sentences, the average term being only 4.5 months.

Selected Topics in Anaerobic Bacteriology.

Science.gov (United States)

Church, Deirdre L

2016-08-01

Alteration in the host microbiome at skin and mucosal surfaces plays a role in the function of the immune system, and may predispose immunocompromised patients to infection. Because obligate anaerobes are the predominant type of bacteria present in humans at skin and mucosal surfaces, immunocompromised patients are at increased risk for serious invasive infection due to anaerobes. Laboratory approaches to the diagnosis of anaerobe infections that occur due to pyogenic, polymicrobial, or toxin-producing organisms are described. The clinical interpretation and limitations of anaerobe recovery from specimens, anaerobe-identification procedures, and antibiotic-susceptibility testing are outlined. Bacteriotherapy following analysis of disruption of the host microbiome has been effective for treatment of refractory or recurrent Clostridium difficile infection, and may become feasible for other conditions in the future.

Cadmium removal by Euglena gracilis is enhanced under anaerobic growth conditions