Oxygen at nanomolar levels reversibly suppresses process rates and gene expression in anammox and denitrification in the oxygen minimum zone off Northern Chil

DEFF Research Database (Denmark)

Dalsgaard, Tage; Stewart, Frank J.; Thamdrup, Bo

2014-01-01

at nanomolar levels affects anammox and denitrification rates and the transcription of nitrogen cycle genes in the anoxic OMZ off Chile. Rates of anammox and denitrification were reversibly suppressed, most likely at the enzyme level. Fiftypercent inhibition of N2 and N2O production by denitrification...

Implication of using different carbon sources for denitrification in wastewater treatments.

Science.gov (United States)

Cherchi, Carla; Onnis-Hayden, Annalisa; El-Shawabkeh, Ibrahim; Gu, April Z

2009-08-01

suggested that the complex biochemical pathways/enzymes involved in the denitrification process depended on the carbon sources used.

Emissions of N2O and CO2, denitrification measurements and soil properties in red clover and ryegrass stands

Czech Academy of Sciences Publication Activity Database

Šimek, Miloslav; Elhottová, Dana; Klimeš, F.; Hopkins, D. W.

2004-01-01

Roč. 36, - (2004), s. 9-21 ISSN 0038-0717 Grant - others:NATO(XX) CRG 97/1331; Evropská unie(XE) EVK2-CT-2000-00096 Institutional research plan: CEZ:AV0Z6066911; CEZ:MSM 122200003 Keywords : emission * denitrification * denitrifying enzyme activity Subject RIV: EH - Ecology, Behaviour Impact factor: 2.234, year: 2004

Distributed denitrification in a northeastern agricultural landscape

Science.gov (United States)

Anderson, T. R.; Groffman, P. M.; Walter, M. T.

2011-12-01

Denitrification may be an important sink of anthropogenic nitrogen (N) in eastern US watersheds. Denitrification occurs primarily under anaerobic conditions by heterotrophic microbes, and is therefore expected to be vigorous in wet soils containing large amounts of organic carbon. Actual rates of denitrification, however, have been difficult to quantify, and remain one of the critical unresolved N processes at the landscape scale. We measured denitrification rates in situ along hydrologic flow paths and across gradients of hydroperiodicities, i.e., frequencies and durations of saturated conditions, at Cornell University's Teaching & Research Center in Harford, NY (an active dairy farm). Denitrification rates were measured monthly using the 15N push-pull method from 14 mini-piezometers arrayed along a gradient of hydroperiodicity as indicated by a soil topographic index (STI). Measured rates of denitrification were spatially variable across sites and ranged from undetectable to over 4500 μg N/kg soil/day with a mean of 572 ± 167 μg N/kg soil/day. Mean rates of denitrification increased with STI, which ranged from 8.7 to 23.0 across our mini-piezometer sites. This relationship was used to estimate denitrification rates across the landscape and resolve a missing piece of the N budget for the farm. Only 14% of the farm fell into areas of STI greater than 8.7; however, denitrification in these areas account for more than 60% of the missing N balance for the entire landscape. Improved understanding of the distribution and magnitudes of denitrification in agricultural landscapes has good potential to facilitate new, novel, and better management practices for controlling N loading to streams and rivers. Indeed, the very areas that appear to have a propensity to harbor denitrification, i.e., areas prone to be wet, are often artificially drained as part of standard agricultural practices which reduces the frequency that these areas are likely to be anaerobic and

Small scale denitrification variability in riparian zones: Results from a high-resolution dataset

Science.gov (United States)

Gassen, Niklas; Knöller, Kay; Musolff, Andreas; Popp, Felix; Lüders, Tillmann; Stumpp, Christine

2017-04-01

Riparian zones are important compartments at the interface between groundwater and surface water where biogeochemical processes like denitrification are often enhanced. Nitrate loads of either groundwater entering a stream through the riparian zone or streamwater infiltrating into the riparian zone can be substantially reduced. These processes are spatially and temporally highly variable, making it difficult to capture solute variabilities, estimate realistic turnover rates and thus to quantify integral mass removal. A crucial step towards a more detailed characterization is to monitor solutes on a scale which adequately resemble the highly heterogeneous distribution and on a scale where processes occur. We measured biogeochemical parameters in a spatial high resolution within a riparian corridor of a German lowland river system over the course of one year. Samples were taken from three newly developed high-resolution multi-level wells with a maximum vertical resolution of 5 cm and analyzed for major ions, DOC and N-O isotopes. Sediment derived during installation of the wells was analyzed for specific denitrifying enzymes. Results showed a distinct depth zonation of hydrochemistry within the shallow alluvial aquifer, with a 1 m thick zone just below the water table with lower nitrate concentrations and EC values similar to the nearby river. Conservative parameters were consistent inbetween the three wells, but nitrate was highly variable. In addition, spots with low nitrate concentrations showed isotopic and microbial evidence for higher denitrification activities. The depth zonation was observed throughout the year, with stronger temporal variations of nitrate concentrations just below the water table compared to deeper layers. Nitrate isotopes showed a clear seasonal trend of denitrification activities (high in summer, low in winter). Our dataset gives new insight into river-groundwater exchange processes and shows the highly heterogeneous distribution of

Syringe test screening of microbial gas production activity: Cases denitrification and biogas formation.

Science.gov (United States)

Østgaard, Kjetill; Kowarz, Viktoria; Shuai, Wang; Henry, Ingrid A; Sposob, Michal; Haugen, Hildegunn Hegna; Bakke, Rune

2017-01-01

Mass produced plastic syringes may be applied as vessels for cheap, simple and large scale batch culture testing. As illustrated for the cases of denitrification and of biogas formation, metabolic activity was monitored by direct reading of the piston movement due to the gas volume formed. Pressure buildup due to friction was shown to be moderate. A piston pull and slide back routine can be applied before recording gas volume to minimize experimental errors due to friction. Inoculum handling and activity may be conveniently standardized as illustrated by applying biofilm carriers. A robust set of positive as well as negative controls ("blanks") should be included to ensure quality of the actual testing. The denitrification test showed saturation response at increasing amounts of inoculum in the form of adapted moving bed biofilm reactor (MBBR) carriers, with well correlated nitrate consumption vs. gas volume formed. As shown, the denitrification test efficiently screened different inocula at standardized substrates. Also, different substrates were successfully screened and compared at standardized inocula. The biogas potential test showed efficient screening of different substrates with effects of relative amounts of carbohydrate, protein, fat. A second case with CO 2 capture reclaimer waste as substrate demonstrated successful use of co-feeding to support waste treatment and how temperature effects on kinetics and stoichiometry can be observed. In total, syringe test screening of microbial gas production seems highly efficient at a low cost when properly applied. Copyright © 2016 Elsevier B.V. All rights reserved.

Oxygen requirement for denitrification by the fungus Fusarium oxysporum.

Science.gov (United States)

Zhou, Z; Takaya, N; Sakairi, M A; Shoun, H

2001-01-01

The effects of dioxygen (O2) on the denitrification activity of the fungus Fusarium oxysporum MT-811 in fed-batch culture in a stirred jar fermentor were examined. The results revealed that fungal denitrifying activity requires a minimal amount of O2 for induction, which is repressed by excess O2. The optimal O2 supply differed between the denitrification substrates : 690 micromol O2 x h(-1) (g dry cell wt.)(-1) for nitrate (NO3-) and about 250 micromol O2 x h(-1) (g dry cell wt.)(-1) for nitrite (NO2-). The reduction of NO3- required more O2 than that of NO2- . With an optimal O2 supply, 80% and 52% of nitrogen atoms in NO3- and NO2-, respectively, were recovered as the denitrification product N2O. These features of F. oxysporum differ from those of bacterial denitrifiers that work exclusively under anoxic conditions. The denitrification activity of F. oxysporum MT-811 mutants with impaired NO3- assimilation was about double that of the wild-type strain, suggesting competition for the substrate between assimilatory and dissimilatory types of NO3- reduction. These results showed that denitrification by F. oxysporum has unique features, namely, a minimal O2 requirement and competition with assimilatory NO3-.

Performance and microbial community analysis of bio-electrocoagulation on simultaneous nitrification and denitrification in submerged membrane bioreactor at limited dissolved oxygen.

Science.gov (United States)

Li, Liang; Dong, Yihua; Qian, Guangsheng; Hu, Xiaomin; Ye, Linlin

2018-06-01

A pair of Fe-C electrodes was installed in a traditional submerged membrane bioreactor (MBR, Rc), and a novel asynchronous periodic reversal bio-electrocoagulation system (Re) was developed. The simultaneous nitrification and denitrification (SND) performance was discussed under limited dissolved oxygen (DO). Results showed that electrocoagulation enhanced total nitrogen (TN) removal from 59.48% to 75.09% at 1.2 mg/L DO. Additionally, Fe electrode could increase sludge concentration, particle size, and enzyme activities related to nitrogen removal. The enzyme activities of Hydroxylamine oxidoreductase (HAO), Nitrate Reductase (NAR), nitric oxide reductase NOR and nitrous oxide reductase (N 2 OR) in Re were 38.35%, 21.59%, 89.96% and 38.64% higher than Rc, respectively. Moreover, electrocoagulation was advantageous for nitrite accumulation, indicating partial nitrification and denitrification were more easily achieved in Re. Besides, results from high throughput sequencing analysis revealed that electrocoagulation increased the relative abundance of most genera related to nitrogen removal, including Nitrosomonas, Comamonadaceae_unclassified, Haliangium and Denitratisoma. Copyright © 2018 Elsevier Ltd. All rights reserved.

Effect of growing plants on denitrification at high soil nitrate concentrations

International Nuclear Information System (INIS)

Haider, K.; Mosier, A.; Heinemeyer, O.

1987-01-01

The availability of plant rhizosphere C deposits and its influence on microbial denitrification is not clearly defined. Conflicting reports as to the influence of plants and root exudation on denitrification continue to appear in the literature. The results of the authors earlier phytotron study indicated that denitrification was not stimulated in soils planted with corn or wheat compared to unplanted soils. Lower nitrate concentrations in the planted soils, however, may have led to misinterpretation of this data. A second study was conducted, to evaluate the effect of actively growing plants on denitrification where the NO 3 7 content of planted soils was maintained similar to unplanted soils. Simultaneously the C fixed by corn (Zea mays) and the fate of fertilizer N applied to the soil during the growing season were quantified. The corn was grown in a phytotron under a continuous supply of 14 CO 2 in 15 N fertilized soils to which 15 N-NO 3 - was added periodically during the growing season. The results of these studies showed that denitrification was not stimulated in soils planted with corn during active plant growth phase even when soil NO 3 - was relatively high. Denitrification was, however, greater in corn planted than unplanted soil when the recoverable root biomass began to decrease. Less N was immobilized and net 15 N immobilization was lower in planted soils than in unplanted soils. As denitrification was lower in planted soils during the time of active plant growth, the study suggests that root exudates did not stimulate either process

Assessment of denitrification process in lower Ishikari river system, Japan.

Science.gov (United States)

Jha, Pawan Kumar; Minagawa, Masao

2013-11-01

Sediment denitrification rate and its role in removal of dissolved nitrate load in lower Ishikari river system were examined. Denitrification rate were measured using acetylene inhibition technique on the sediment samples collected during August 2009-July 2010. The denitrification rate varied from 0.001 to 1.9 μg Ng(-1) DM h(-1) with an average value of 0.21 μg Ng(-1) DM h(-1) in lower Ishikari river system. Denitrification rate showed positive correlation with dissolved nitrate concentration in the river basin, indicating overlying water column supplied nitrate for the sediment denitrification processes. Nutrient enrichment experiments result showed that denitrification rate increased significantly with addition of nitrate in case of samples collected from Barato Lake however no such increase was observed in the samples collected from Ishikari river main channel and its major tributaries indicating that factors other than substrate concentration such as population of denitrifier and hydrological properties of stream channel including channel depth and flow velocity may affects the denitrification rate in lower Ishikari river system. Denitrification rate showed no significant increase with the addition of labile carbon (glucose), indicating that sediment samples had sufficient organic matter to sustain denitrification activity. The result of nutrient spiraling model indicates that in- stream denitrification process removes on an average 5%d(-1) of dissolve nitrate load in Ishikari river. This study was carried out to fill the gap present in the availability of riverine denitrification rate measurement and its role in nitrogen budget from Japanese rivers characterize by small river length and high flow rate. Copyright © 2013 Elsevier Ltd. All rights reserved.

Denitrification across landscapes and waterscapes: A synthesis

Science.gov (United States)

Seitzinger, S.; Harrison, J.A.; Böhlke, J.K.; Bouwman, A.F.; Lowrance, R.; Peterson, B.; Tobias, C.; Van Drecht, G.

2006-01-01

Denitrification is a critical process regulating the removal of bioavailable nitrogen (N) from natural and human-altered systems. While it has been extensively studied in terrestrial, freshwater, and marine systems, there has been limited communication among denitrification scientists working in these individual systems. Here, we compare rates of denitrification and controlling factors across a range of ecosystem types. We suggest that terrestrial, freshwater, and marine systems in which denitrification occurs can be organized along a continuum ranging from (1) those in which nitrification and denitrification are tightly coupled in space and time to (2) those in which nitrate production and denitrification are relatively decoupled.In aquatic ecosystems, N inputs influence denitrification rates whereas hydrology and geomorphology influence the proportion of N inputs that are denitrified. Relationships between denitrification and water residence time and N load are remarkably similar across lakes, river reaches, estuaries, and continental shelves.Spatially distributed global models of denitrification suggest that continental shelf sediments account for the largest portion (44%) of total global denitrification, followed by terrestrial soils (22%) and oceanic oxygen minimum zones (OMZs; 14%). Freshwater systems (groundwater, lakes, rivers) account for about 20% and estuaries 1% of total global denitrification. Denitrification of land-based N sources is distributed somewhat differently. Within watersheds, the amount of land-based N denitrified is generally highest in terrestrial soils, with progressively smaller amounts denitrified in groundwater, rivers, lakes and reservoirs, and estuaries. A number of regional exceptions to this general trend of decreasing denitrification in a downstream direction exist, including significant denitrification in continental shelves of N from terrestrial sources. Though terrestrial soils and groundwater are responsible for much

Denitrification in the water column of the central Baltic Sea

DEFF Research Database (Denmark)

Dalsgaard, Tage; De Brabandere, Loreto; Hall, Per O. J.

2013-01-01

Removal of fixed nitrogen in the water column of the eastern Gotland Basin, central Baltic Sea, was studied during two cruises in September 2008 and August 2010. The water column was stratified with anoxic sulfidic bottom water meeting oxic nitrate containing water at the oxic–anoxic interface......3 or sulfide concentrations were converted to in situ rates using the measured water column concentrations of NO3 and sulfide and the actual measured relations between NO3 and sulfide concentrations and enitrification rates. In situ denitrification ranged from 0.24 to 15.9 nM N2 h1. Assuming...... area. Even with an active denitrification layer of 3–6 m thickness the pelagic denitrification per unit area clearly exceeded sediment denitrification rates elsewhere in the Baltic Sea. When extrapolated to the entire Baltic Proper (BP) denitrification in the water column was in the range of 132...

Denitrification

National Research Council Canada - National Science Library

Payne, W. J

1981-01-01

.... After an introduction on the discovery of the denitrification process and the development of research into it, the author considers successively the four anerobic, reductive steps that comprise...

Denitrification prevails over anammox in tropical mangrove sediments (Goa, India)

Digital Repository Service at National Institute of Oceanography (India)

Fernandes, S.O.; Michotey, V.D.; Guasco, S.; Bonin, P.C.; LokaBharathi, P.A.

denitrification as the main process of N sub(2) production in mangrove sediments. At Divar, denitrification was approx 3 times higher than at Tuvem with maximum activity of 224.51 + or -6.63 nmol N sub(2) g sup(−1) h sup(−1) at 0–2 cm. Denitrifying genes (nos...

Potential Denitrification: Concept and Conditions of its Measurement ...

African Journals Online (AJOL)

Denitrification plays important roles in nitrogen cycling and management affecting both the environment and agricultural systems. Potential denitrification has a role in comparison of denitrification magnitudes in different soils. This paper discusses the concept of potential denitrification in relation to denitrification rate and ...

Advances on simultaneous desulfurization and denitrification using activated carbon irradiated by microwaves.

Science.gov (United States)

Ma, Shuang-Chen; Gao, Li; Ma, Jing-Xiang; Jin, Xin; Yao, Juan-Juan; Zhao, Yi

2012-06-01

This paper describes the research background and chemistry of desulfurization and denitrification technology using microwave irradiation. Microwave-induced catalysis combined with activated carbon adsorption and reduction can reduce nitric oxide to nitrogen and sulfur dioxide to sulfur from flue gas effectively. This paper also highlights the main drawbacks of this technology and discusses future development trends. It is reported that the removal of sulfur dioxide and nitric oxide using microwave irradiation has broad prospects for development in the field of air pollution control.

Modelling the soil nitrogen denitrification

International Nuclear Information System (INIS)

Budoi, G.H.; Danuso, F.; Giovanardi, R.; Gavriluta, A.; Alexandrescu, A.; Bireescu, L.

1999-01-01

The paper presents the differential equations used to compute the daily amounts of N denitrified and to compute the amount of N denitrified in a given period of time. It shows also the equations which compute the correction factors of the maximum denitrification rate as a function of soil temperature (F td ), moisture (F md ) and pH (F pHd ), original equations used by NICROS - nitrogen crop simulation model to describe the influence of these abiotic factors. The temperature factor, F td . The optimum temperature for denitrification is between 25-37 o C. The process is slow at temperatures below 10 o C, there is an increased inhibition below 5 o C and stop completely at 0 o C. The maximum temperature for denitrification is practically that which limits the soil microbiological activity, generally 75 o C. The following relations are used to compute the F td factor: F td 1/(1 + e -0,3347 tmed+ 4,99 ) if t med ≤ 37; F td = 1 - (t med - 37)/38 if 75 > t med > 37; F td = 0 if t med ≥ 75, where t med is the average daily soil temperature. The moisture factor, F md . The denitrification has maximum intensity at soil water saturation, U sat , and stop below 80 % from U sat . F md = 0 if soil moisture U s ≤ 0,8*U sat , and F md = (U s - 0,8*U sat )/(U sat - 0,8*U sat ) if U s > 0,8*U sat . The pH factor, F pHd . Denitrification takes place at pH between 4-9 and is maximum at pH between 7-8. The relations used to compute the F pHd factor are: F pHd = 1/(1 + e -3,1923 pH + 18,87 ) if pH ≤ 8; F pHd = (9 - pH) when pH is between 8-9, and F pHd = 0 if pH > 9. Refs. 6 (author)

Denitrification processes in the Arabian Sea

Digital Repository Service at National Institute of Oceanography (India)

Naqvi, S.W.A.

rate in the vicinity of 30 Tg Ny@u-1@@, but the extent of benthic contribution remains unknown. A decoupling of denitrification from primary production, unique to the Arabian Sea, is revealed by nitrite, Electron Transport System (ETS) activity...

Bacterial community evolutions driven by organic matter and powder activated carbon in simultaneous anammox and denitrification (SAD) process.

Science.gov (United States)

Ge, Cheng-Hao; Sun, Na; Kang, Qi; Ren, Long-Fei; Ahmad, Hafiz Adeel; Ni, Shou-Qing; Wang, Zhibin

2018-03-01

A distinct shift of bacterial community driven by organic matter (OM) and powder activated carbon (PAC) was discovered in the simultaneous anammox and denitrification (SAD) process which was operated in an anti-fouling submerged anaerobic membrane bio-reactor. Based on anammox performance, optimal OM dose (50 mg/L) was advised to start up SAD process successfully. The results of qPCR and high throughput sequencing analysis indicated that OM played a key role in microbial community evolutions, impelling denitrifiers to challenge anammox's dominance. The addition of PAC not only mitigated the membrane fouling, but also stimulated the enrichment of denitrifiers, accounting for the predominant phylum changing from Planctomycetes to Proteobacteria in SAD process. Functional genes forecasts based on KEGG database and COG database showed that the expressions of full denitrification functional genes were highly promoted in R C , which demonstrated the enhanced full denitrification pathway driven by OM and PAC under low COD/N value (0.11). Copyright © 2017 Elsevier Ltd. All rights reserved.

Nitric oxide in denitrification - an elusive signal molecule emitted from soil

Science.gov (United States)

Bakken, L. R.; Frostegard, A.

2010-12-01

Soils emit variable amounts of NO and N2O, with environmental consequences (atmosphere chemistry and global warming). Nitrification was for some time considered the main source of NO emission, but several investigations have indicated that denitrification may be a potent source as well. However, strong emission of NO from denitrifying organisms is in some conflict with common understanding of the role of NO in the regulation of denitrification, as based on paradigm model strains. NO appears to be an important signal molecule for denitrifying organisms by exerting a positive feedback on the expression of the genes coding for denitrification. On the other hand, a careful control of the NO concentrations at nanomolar concentrations has long been considered an essential fitness character for denitrifying organisms, since micromolar concentrations of NO is toxic to many organisms. For the same reason, organisms lacking genes encoding NO reductase (NOR) have been considered unfit for denitrification. This view is challenged by isolation of organisms whose primary product of denitrification is NO, either because they lack the genes for NO reductase, or because their synthesis of the denitrification proteome is extremely unbalanced, resulting in transient NO accumulation to micromolar concentrations when grown in pure culture. Such paralyzing NO concentrations are probably never reached in natural environments, however, due to diffusion and NO-absorption by adjacent organisms, be it by NOR or other NO scavenging enzymes. Hypothetically, the production of NO by denitrifying organisms may be an advantage by fending off nearby competitors. We have embarked on a comparative study of denitrification phenotypes regarding their denitrification gene expression and control of NO and N2O concentrations in response to anoxic spells. This includes model strains (Paracoccus denitrificans and Agrobacterium tumefaciens) and recently isolated strains within several genera. Some are found

Denitrification as an adaptive trait in soil and groundwater bacteria

Energy Technology Data Exchange (ETDEWEB)

Bergwall, C

1997-09-01

The focus of this thesis is on selection and adaptation processes in bacteria with emphasis on denitrifying bacteria in groundwater. Other nitrogen transformation processes such as dissimilatory nitrate reduction to ammonium (nitrate ammonification) and nitrification of forest soil bacteria are briefly discussed. Microcosms with sterile sediment and groundwater were inoculated with single denitrifying strains isolated from three groundwater aquifers, two of which are agricultural aquifers (in situ NO{sub 3}{sup -}-N was 24.1 and 35.2 mg1{sup -1}) and the third which is a pristine lake water infiltration aquifer (in situ NO{sub 3}{sup -}-N was 6.3 mg1{sup -1}). The average denitrification activity for strains from the nitrate contaminated sites were twice as high as the activity of the strains from the pristine site. Denitrification were carbon limited and glucose amendment increased the denitrification activity about a 2-fold for all strains. The strain specific differences in denitrification rates increased to a 2.5-fold after carbon addition indicating that the differences in reduction rates cannot be explained by different carbon utilisation rates but rather reflect innate differences in the reductases of the strains. A preliminary identification of the molecular target for adaptation was performed with artificial electron donors and electron acceptors for all enzymatic steps in the denitrification pathway. Nitrous oxide reductase activity was significantly higher in denitrifiers from the nitrate contaminated sites. This suggests that nos genes may be the molecular target, possibly by mutation or gene duplication for adaptation to high nitrate concentrations. Two anaerobic denitrifiers from each of the contaminated sites were capable of aerobic denitrification indicating that high nitrate concentrations may select for strains that denitrifies in the presence of both oxygen and nitrate. Microcosm experiments with fertilized coniferous forest soil showed that the

Denitrification gene expression in clay-soil bacterial community

Science.gov (United States)

Pastorelli, R.; Landi, S.

2009-04-01

Our contribution in the Italian research project SOILSINK was focused on microbial denitrification gene expression in Mediterranean agricultural soils. In ecosystems with high inputs of nitrogen, such as agricultural soils, denitrification causes a net loss of nitrogen since nitrate is reduced to gaseous forms, which are released into the atmosphere. Moreover, incomplete denitrification can lead to emission of nitrous oxide, a potent greenhouse gas which contributes to global warming and destruction of ozone layer. A critical role in denitrification is played by microorganisms and the ability to denitrify is widespread among a variety of phylogenetically unrelated organisms. Data reported here are referred to wheat cultivation in a clay-rich soil under different environmental impact management (Agugliano, AN, Italy). We analysed the RNA directly extracted from soil to provide information on in situ activities of specific populations. The expression of genes coding for two nitrate reductases (narG and napA), two nitrite reductases (nirS and nirK), two nitric oxide reductases (cnorB and qnorB) and nitrous oxide reductase (nosZ) was analyzed by reverse transcription (RT)-nested PCR. Only napA, nirS, nirK, qnorB and nosZ were detected and fragments sequenced showed high similarity with the corresponding gene sequences deposited in GenBank database. These results suggest the suitability of the method for the qualitative detection of denitrifying bacteria in environmental samples and they offered us the possibility to perform the denaturing gradient gel electrophoresis (DGGE) analyzes for denitrification genes.. Earlier conclusions showed nirK gene is more widely distributed in soil environment than nirS gene. The results concerning the nosZ expression indicated that microbial activity was clearly present only in no-tilled and no-fertilized soils.

Denitrification potential of riparian soils in relation to multiscale spatial environmental factors: a case study of a typical watershed, China.

Science.gov (United States)

Wei, Jianbing; Feng, Hao; Cheng, Quanguo; Gao, Shiqian; Liu, Haiyan

2017-02-01

The objective of this study was to test the hypothesis that environmental regulators of riparian zone soil denitrification potential differ according to spatial scale within a watershed; consequently, a second objective was to provide spatial strategies for conserving and restoring the purification function of runoff in riparian ecosystems. The results show that soil denitrification in riparian zones was more heterogeneous at the profile scale than at the cross-section and landscape scales. At the profile scale, biogeochemical factors (including soil total organic carbon, total nitrogen, and nitrate-nitrogen) were the major direct regulators of the spatial distribution of soil denitrification enzyme activity (DEA). At the cross-section scale, factors included distance from river bank and vegetation density, while landscape-scale factors, including topographic index, elevation, and land use types, indirectly regulated the spatial distribution of DEA. At the profile scale, soil DEA was greatest in the upper soil layers. At the cross-section scale, maximum soil DEA occurred in the mid-part of the riparian zone. At the landscape scale, soil DEA showed an increasing trend towards downstream sites, except for those in urbanized areas.

Quantification and enzyme targets of fatty acid amides from duckweed root exudates involved in the stimulation of denitrification.

Science.gov (United States)

Sun, Li; Lu, Yufang; Kronzucker, Herbert J; Shi, Weiming

2016-07-01

Fatty acid amides from plant root exudates, such as oleamide and erucamide, have the ability to participate in strong plant-microbe interactions, stimulating nitrogen metabolism in rhizospheric bacteria. However, mechanisms of secretion of such fatty acid amides, and the nature of their stimulatory activities on microbial metabolism, have not been examined. In the present study, collection, pre-treatment, and determination methods of oleamide and erucamide in duckweed root exudates are compared. The detection limits of oleamide and erucamide by gas chromatography (GC) (10.3ngmL(-1) and 16.1ngmL(-1), respectively) are shown to be much lower than those by liquid chromatography (LC) (1.7 and 5.0μgmL(-1), respectively). Quantitative GC analysis yielded five times larger amounts of oleamide and erucamide in root exudates of Spirodela polyrrhiza when using a continuous collection method (50.20±4.32 and 76.79±13.92μgkg(-1) FW day(-1)), compared to static collection (10.88±0.66 and 15.27±0.58μgkg(-1) FW day(-1)). Furthermore, fatty acid amide secretion was significantly enhanced under elevated nitrogen conditions (>300mgL(-1)), and was negatively correlated with the relative growth rate of duckweed. Mechanistic assays were conducted to show that erucamide stimulates nitrogen removal by enhancing denitrification, targeting two key denitrifying enzymes, nitrate and nitrite reductases, in bacteria. Our findings significantly contribute to our understanding of the regulation of nitrogen dynamics by plant root exudates in natural ecosystems. Copyright © 2016 Elsevier GmbH. All rights reserved.

Copepod carcasses as microbial hot spots for pelagic denitrification

DEFF Research Database (Denmark)

Glud, Ronnie N.; Grossart, Hans-Peter; Larsen, Morten

2015-01-01

Copepods are exposed to a high non-predatory mortality and their decomposing carcasses act as microniches with intensified microbial activity. Sinking carcasses could thereby represent anoxic microenvironment sustaining anaerobic microbial pathways in otherwise oxic water columns. Using non...... investigated carcass samples and thereby documented the potential for microbial denitrification in carcasses. The nirS gene was occasionally expressed in live copepods, but not as consistently as in carcasses. Incubations of sinking carcasses in 15NO2 3 amended seawater demonstrated denitrification, of which...

Measuring the Enzyme Activity of Arabidopsis Deubiquitylating Enzymes.

Science.gov (United States)

Kalinowska, Kamila; Nagel, Marie-Kristin; Isono, Erika

2016-01-01

Deubiquitylating enzymes, or DUBs, are important regulators of ubiquitin homeostasis and substrate stability, though the molecular mechanisms of most of the DUBs in plants are not yet understood. As different ubiquitin chain types are implicated in different biological pathways, it is important to analyze the enzyme characteristic for studying a DUB. Quantitative analysis of DUB activity is also important to determine enzyme kinetics and the influence of DUB binding proteins on the enzyme activity. Here, we show methods to analyze DUB activity using immunodetection, Coomassie Brilliant Blue staining, and fluorescence measurement that can be useful for understanding the basic characteristic of DUBs.

Treatment of domestic wastewater by anaerobic denitrification ...

African Journals Online (AJOL)

SALAMA YOUSSEF

for the denitrification process could be deduced from the. pH variation. The alkalinity and pH increased in heterotrophic and H2-based autotrophic denitrification because nitrite reduction consumed protons (H+). Proton consumption is illustrated in Equations (1 to 4) (Rittmann and McCarty, 2001). Heterotrophic denitrification.

Transcriptional and metabolic regulation of denitrification in Paracoccus denitrificans allows low but significant activity of nitrous oxide reductase under oxic conditions.

Science.gov (United States)

Qu, Zhi; Bakken, Lars R; Molstad, Lars; Frostegård, Åsa; Bergaust, Linda L

2016-09-01

Oxygen is known to repress denitrification at the transcriptional and metabolic levels. It has been a common notion that nitrous oxide reductase (N2 OR) is the most sensitive enzyme among the four N-oxide reductases involved in denitrification, potentially leading to increased N2 O production under suboxic or fluctuating oxygen conditions. We present detailed gas kinetics and transcription patterns from batch culture experiments with Paracoccus denitrificans, allowing in vivo estimation of e(-) -flow to O2 and N2 O under various O2 regimes. Transcription of nosZ took place concomitantly with that of narG under suboxic conditions, whereas transcription of nirS and norB was inhibited until O2 levels approached 0 μM in the liquid. Catalytically functional N2 OR was synthesized and active in aerobically raised cells transferred to vials with 7 vol% O2 in headspace, but N2 O reduction rates were 10 times higher when anaerobic pre-cultures were subjected to the same conditions. Upon oxygen exposure, there was an incomplete and transient inactivation of N2 OR that could be ascribed to its lower ability to compete for electrons compared with terminal oxidases. The demonstrated reduction of N2 O at high O2 partial pressure and low N2 O concentrations by a bacterium not known as a typical aerobic denitrifier may provide one clue to the understanding of why some soils appear to act as sinks rather than sources for atmospheric N2 O. © 2015 Society for Applied Microbiology and John Wiley & Sons Ltd.

Flood-controlled excess-air formation favors aerobic respiration and limits denitrification activity in riparian groundwater

Directory of Open Access Journals (Sweden)

Simone ePeter

2015-11-01

Full Text Available The saturated riparian zones of rivers act as spatially and temporally variable biogeochemical reactors. This complicates the assessment of biogeochemical transport and transformation processes. During a flood event, excess-air formation, i.e. the inclusion and dissolution of air bubbles into groundwater, can introduce high amounts of dissolved O2 and thereby affect biogeochemical processes in groundwater. With the help of a field-installed membrane-inlet mass-spectrometer we resolved the effects of flood induced excess-air formationon organic carbon and nitrogen transformations in groundwater of different riparian zones of a restored section of the River Thur, Switzerland. The results show that the flood event triggered high aerobic respiration activity in the groundwater below a zone densely populated with willow plants. The flood introduced high concentrations of O2 (230 µmol L–1 to the groundwater through the formation of excess air and transported up to ~400 µmol L 1 organic carbon from the soil/root layer into groundwater during the movement of the water table. A rapid respiration process, quantified via the measurements of O2, CO2 and noble-gas concentrations, led to fast depletion of the introduced O2 and organic carbon and to high CO2 concentration (590 µmol L–1 in the groundwater shortly after the flood. The synchronous analysis of different nitrogen species allowed studying the importance of denitrification activity. The results indicate that in the willow zone excess-air formation inhibited denitrification through high O2 concentration input. Instead, the observed decrease in nitrate concentration (~50 µmol N L 1 may be related to fostered nitrate uptake by plants. In the other riparian zones closer to the river, no significant excess-air formation and corresponding respiration activity was observed. Overall, analyzing the dissolved gases in the groundwater significantly contributed to deciphering biogeochemical processes in

Regulation and role of epiphytic nitrification and denitrification in macrophyte-dominated systems

Energy Technology Data Exchange (ETDEWEB)

Eriksson, Peder G.

2000-02-01

This thesis examines mechanisms regulating bacterial nitrification and denitrification in attached microbial communities on surfaces of aquatic macrophytes. It also evaluates the role of epiphytic nitrification and denitrification for the nitrogen turnover in macrophyte-dominated nutrient-rich freshwater. Epiphytic nitrification is promoted in light and epiphytic denitrification occurs mainly in dark, because the metabolic activity of the aquatic macrophyte and its epiphytes induce in light high and in dark low oxygen concentrations in epiphytic communities. Epiphytic nitrification and denitrification are also affected by the physical and chemical characteristics of the aquatic macrophyte. The spatial distribution of nitrification in emergent macrophyte wetlands is related to the species composition of the emergent vegetation, possibly because of a macrophyte species-related release of organic nitrification inhibitors. Contrasting to nitrifying bacteria, which are lithotrophic, denitrifying bacteria use organic substances as an energy source and are therefore stimulated by the release of organic matter from aquatic macrophytes. Epiphytic communities support more denitrification in nutrient-rich than in nutrient-poor environments. In lakes and ponds, epiphytic denitrification is higher at sheltered locations than at locations exposed to wind-induced water movements or currents. In flowing water, epiphytic denitrification occurs mainly at low oxygen concentrations in the surrounding water. However, because aquatic macrophytes impede water flow and induce low oxygen concentrations in dark, epiphytic denitrification can be present within dense vegetation despite of high oxygen concentrations in the surrounding water. Epiphytic nitrification is almost unaffected by flow conditions, and can occur both in light and in dark. In shallow-water systems such as treatment wetlands, aquatic macrophytes often provide most of the accessible surface area for attached nitrifying and

Studies on denitrification performance of tricking filters

International Nuclear Information System (INIS)

Ruediger, A.

1993-01-01

Trickling filters are one of the oldest methods of biological waste water purification, widely used ever since they were first developed. These filters are mostly used for aerobic purification of water as highly polluted or lightly polluted reactors. While these systems are very useful for the elemination of organic pollution and the nitrification of the waste waters, denitrification of the whole plant poses considerable problems. The question is in how far trickling filters can be used as denitrification reactors. The conditions of successful denitrification in trickling filters is investigated, denitrification performance is established. Studies were conducted in laboratory, semi-industrial and industrial scale. (BBR) [de

Denitrification in Soil Aggregate Analogues-Effect of Aggregate Size and Oxygen Diffusion

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Steffen Schlüter

2018-04-01

Full Text Available Soil-borne nitrous oxide (N2O emissions have a high spatial and temporal variability which is commonly attributed to the occurrence of hotspots and hot moments for microbial activity in aggregated soil. Yet there is only limited information about the biophysical processes that regulate the production and consumption of N2O on microscopic scales in undisturbed soil. In this study, we introduce an experimental framework relying on simplified porous media that circumvents some of the complexities occuring in natural soils while fully accounting for physical constraints believed to control microbial activity in general and denitrification in particular. We used this framework to explore the impact of aggregate size and external oxygen concentration on the kinetics of O2 consumption, as well as CO2 and N2O production. Model aggregates of different sizes (3.5 vs. 7 mm diameter composed of porous, sintered glass were saturated with a defined growth medium containing roughly 109 cells ml−1 of the facultative anaerobic, nosZ-deficient denitrifier Agrobacterium tumefaciens with N2O as final denitrification product and incubated at five different oxygen levels (0–13 vol-%. We demonstrate that the onset of denitrification depends on the amount of external oxygen and the size of aggregates. Smaller aggregates were better supplied with oxygen due to a larger surface-to-volume ratio, which resulted in faster growth and an earlier onset of denitrification. In larger aggregates, the onset of denitrification was more gradual, but with comparably higher N2O production rates once the anoxic aggregate centers were fully developed. The normalized electron flow from the reduced carbon substrate to N-oxyanions (edenit-/etotal- ratio could be solely described as a function of initial oxygen concentration in the headspace with a simple, hyperbolic model, for which the two empirical parameters changed with aggregate size in a consistent way. These findings confirm the

Assessment of the denitrification process in alluvial wetlands at floodplain scale using the SWAT model

Science.gov (United States)

As alluvial plains support intensive agricultural activities, they often suffer from groundwater nitrate pollution. Denitrification is recognized as an important process in nitrate pollution control in riparian zones. In shallow aquifer zones influenced by recharged surface water, denitrification ...

Denitrification of acid wastes from uranium purification processes

International Nuclear Information System (INIS)

Clark, F.E.; Francis, C.W.; Francke, H.C.; Strohecker, J.W.

1975-11-01

Laboratory and pilot-plant investigations have shown the technical feasibility of removing nitrates from neutralized acid wastes from uranium purification processes by biological denitrification, a dissimilatory process in which the nitrate ion is reduced to nitrogen gas by specific bacteria. The process requires anaerobic conditions and an organic carbon source, as well as other life-sustaining constituents. These denitrification studies produced process design information on a columnar denitrification plant and on continuous-flow, stirred-bed reactors. Denitrification, using packed columns, was found to be desirable for soluble salts, such as those of sodium and ammonium; denitrification, using stirred reactors, was found to be desirable for mixtures containing insoluble salts, such as those of calcium and aluminum. Packed columns were found to have denitrification rates ranging up to 122 grams of nitrate per day per cubic decimeter of column volume; stirred-bed reactors have been shown to have reaction rates near 10 grams of nitrate per day per cubic decimeter of reactor volume. The continuous-flow, stirred-bed reactors were selected for scaleup studies because of the solids-removal problems associated with packed columns when operating on feeds containing high concentrations of insoluble salts or ions which form insoluble salts with the products of the denitrification reaction

Denitrification in human dental plaque

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Verstraete Willy

2010-03-01

Full Text Available Abstract Background Microbial denitrification is not considered important in human-associated microbial communities. Accordingly, metabolic investigations of the microbial biofilm communities of human dental plaque have focused on aerobic respiration and acid fermentation of carbohydrates, even though it is known that the oral habitat is constantly exposed to nitrate (NO3- concentrations in the millimolar range and that dental plaque houses bacteria that can reduce this NO3- to nitrite (NO2-. Results We show that dental plaque mediates denitrification of NO3- to nitric oxide (NO, nitrous oxide (N2O, and dinitrogen (N2 using microsensor measurements, 15N isotopic labelling and molecular detection of denitrification genes. In vivo N2O accumulation rates in the mouth depended on the presence of dental plaque and on salivary NO3- concentrations. NO and N2O production by denitrification occurred under aerobic conditions and was regulated by plaque pH. Conclusions Increases of NO concentrations were in the range of effective concentrations for NO signalling to human host cells and, thus, may locally affect blood flow, signalling between nerves and inflammatory processes in the gum. This is specifically significant for the understanding of periodontal diseases, where NO has been shown to play a key role, but where gingival cells are believed to be the only source of NO. More generally, this study establishes denitrification by human-associated microbial communities as a significant metabolic pathway which, due to concurrent NO formation, provides a basis for symbiotic interactions.

The mechanism of oxygen isotopic fractionation during fungal denitrification - A pure culture study

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Wrage-Moennig, Nicole; Rohe, Lena; Anderson, Traute-Heidi; Braker, Gesche; Flessa, Heinz; Giesemann, Annette; Lewicka-Szczebak, Dominika; Well, Reinhard

2014-05-01

Nitrous oxide (N2O) from soil denitrification originates from bacteria and - to an unknown extent - also from fungi. During fungal denitrification, oxygen (O) exchange takes place between H2O and intermediates of the denitrification process as in bacterial exchange[1,2]. However, information about enzymes involved in fungal O exchanges and the associated fractionation effects is lacking. The objectives of this study were to estimate the O fractionation and O exchange during the fungal denitrifying steps using a conceptual model[2] adapted from concepts for bacterial denitrification[3], implementing controls of O exchange proposed by Aerssens, et al.[4] and using fractionation models by Snider et al.[5] Six different pure fungal cultures (five Hypocreales, one Sordariales) known to be capable of denitrification were incubated under anaerobic conditions, either with nitrite or nitrate. Gas samples were analyzed for N2O concentration and its isotopic signatures (SP, average δ15N, δ18O). To investigate O exchange, both treatments were also established with 18O-labelled water as a tracer in the medium. The Hypocreales strains showed O exchange mainly at NO2- reductase (Nir) with NO2- as electron acceptor and no additional O exchange at NO3- reductase (Nar) with NO3- as electron acceptor. The only Hypocreales species having higher O exchange with NO3- than with NO2- also showed O exchange at Nar. The Sordariales species tested seems capable of O exchange at NO reductase (Nor) additionally to O exchange at Nir with NO2-. The data will help to better interpret stable isotope values of N2O from soils. .[1] D. M. Kool, N. Wrage, O. Oenema, J. Dolfing, J. W. Van Groenigen. Oxygen exchange between (de)nitrification intermediates and H2O and its implications for source determination of NO?3- and N2O: a review. Rapid Commun. Mass Spec. 2007, 21, 3569. [2] L. Rohe, T.-H. Anderson, B. Braker, H. Flessa, A. Giesemann, N. Wrage-Mönnig, R. Well. Fungal Oxygen Exchange between

Carbon source recovery from excess sludge by mechanical disintegration for biological denitrification.

Science.gov (United States)

Zubrowska-Sudol, M

2018-04-01

The goal of the study was to evaluate the possibility of carbon source recovery from excess sludge by mechanical disintegration for biological denitrification. The total efficiency of denitrification, unit demand for organic compounds for denitrification, unit volume of disintegrated sludge and unit cost of nitrogen removal as a function of energy density used for excess sludge disintegration (70, 140 and 210 kJ/L) were analyzed. In the study a full-scale disc disintegrator was used (motor power: 30 kWh, motor speed: 2,950 rpm). It was shown that the amounts of organic compounds released from the activated sludge flocs at all tested levels of energy density are high enough to be used to intensify the removal of nitrogen compounds from wastewater. It was also documented that the energy density provided during process of disintegration was an important factor determining the characteristics of organic compounds obtained under the disintegration for their use in order to intensify the process of denitrification. The highest value of total efficiency of denitrification (50.5 ± 3.1 mg N/L) was obtained for carbon source recovery from excess sludge at 70 kJ/L, but the lowest unit cost of nitrogen removal occurred for 140 kJ/L (0.0019 ± 0.0011 EUR/g N).

Denitrification activity is closely linked to the total ambient Fe concentration in mangrove sediments of Goa, India

Science.gov (United States)

Fernandes, Sheryl Oliveira; Gonsalves, Maria-Judith; Michotey, Valérie D.; Bonin, Patricia C.; Loka, A.; Bharathi, P.

2013-10-01

Denitrification activity (DNT) and associated environmental parameters were examined in two mangrove ecosystems of Goa, India - the relatively unimpacted Tuvem and the anthropogenically-influenced Divar. Sampling was carried out at every 2 cm interval within the 0-10 cm depth range to determine (1) seasonal (pre-monsoon, monsoon and post-monsoon) down-core variation in DNT (2) assess the environmental factors influencing the DNT and (3) to build predictive models for benthic DNT. Denitrification generally decreased with depth and showed marked seasonal variation at both the locations. Denitrification peaked during the pre-monsoon occurring at a rate of up to 21.00 ± 12.84 nmol N2O g-1 h-1 within 0-4 cm at both the locations. Further, DNT at pre-monsoon was significantly influenced by Fe content at Tuvem and Divar suggesting Fe-mediated nitrate respiration. The influence of other limiting substrates such as NO3- and NO2- was most important during the monsoon and post-monsoon especially at Divar. The multiple regression models developed could predict 67-98% of the observed variability in DNT through the seasons. About 6-9 environmental variables were required to relatively well-predict DNT in these sediments with the complexity governing DNT decreasing from pre-monsoon to post-monsoon. Our results reveal that seasonal dynamics of DNT in tropical mangrove sediments are closely linked to the total Fe at the prevailing ambient concentration in both the systems.

Monovalent Cation Activation of the Radical SAM Enzyme Pyruvate Formate-Lyase Activating Enzyme.

Science.gov (United States)

Shisler, Krista A; Hutcheson, Rachel U; Horitani, Masaki; Duschene, Kaitlin S; Crain, Adam V; Byer, Amanda S; Shepard, Eric M; Rasmussen, Ashley; Yang, Jian; Broderick, William E; Vey, Jessica L; Drennan, Catherine L; Hoffman, Brian M; Broderick, Joan B

2017-08-30

Pyruvate formate-lyase activating enzyme (PFL-AE) is a radical S-adenosyl-l-methionine (SAM) enzyme that installs a catalytically essential glycyl radical on pyruvate formate-lyase. We show that PFL-AE binds a catalytically essential monovalent cation at its active site, yet another parallel with B 12 enzymes, and we characterize this cation site by a combination of structural, biochemical, and spectroscopic approaches. Refinement of the PFL-AE crystal structure reveals Na + as the most likely ion present in the solved structures, and pulsed electron nuclear double resonance (ENDOR) demonstrates that the same cation site is occupied by 23 Na in the solution state of the as-isolated enzyme. A SAM carboxylate-oxygen is an M + ligand, and EPR and circular dichroism spectroscopies reveal that both the site occupancy and the identity of the cation perturb the electronic properties of the SAM-chelated iron-sulfur cluster. ENDOR studies of the PFL-AE/[ 13 C-methyl]-SAM complex show that the target sulfonium positioning varies with the cation, while the observation of an isotropic hyperfine coupling to the cation by ENDOR measurements establishes its intimate, SAM-mediated interaction with the cluster. This monovalent cation site controls enzyme activity: (i) PFL-AE in the absence of any simple monovalent cations has little-no activity; and (ii) among monocations, going down Group 1 of the periodic table from Li + to Cs + , PFL-AE activity sharply maximizes at K + , with NH 4 + closely matching the efficacy of K + . PFL-AE is thus a type I M + -activated enzyme whose M + controls reactivity by interactions with the cosubstrate, SAM, which is bound to the catalytic iron-sulfur cluster.

Denitrification in groundwater at uranium mill tailings sites

International Nuclear Information System (INIS)

Goering, Timothy J.; Groffman, Armando; Thomson, Bruce

1992-01-01

Nitrates are a major contaminant in groundwater at many Uranium Mill Tailings Remedial Action (UMTRA) sites. Microbial denitrification, the transformation of nitrate to nitrogen gas, may be occurring in groundwater at several UMTRA sites. Denitrification is a biologically mediated process whereby facultative anaerobes use nitrate for respiration under anaerobic conditions. Denitrifying bacteria are ubiquitous in soils, sediments, and water. Denitrification requires nitrate, organic carbon, oxygen-limiting conditions, and trace nutrients, especially phosphorus. The lack of organic carbon is the most common limiting factor for denitrification. Denitrification occurs under a limited range of temperature and pH. The uranium milling processes used at UMTRA sites provided a readily available source of carbon and nitrates for denitrifying bacteria. At the Maybell, Colorado, site, the denitrifying organisms Pseudomonas, Flavobacterium and Acinetobacter were identified in core samples of materials from beneath the tailings. In addition, microcosm experiments simulating aquifer conditions beneath the tailings pile showed an average 40 percent decrease in nitrate concentrations over 13 days. At the New Rifle, Colorado, site, aquifer conditions appear favorable for denitrification. Nitrate and organic carbon are readily available in the groundwater, and redox conditions beneath and downgradient of the tailings pile are relatively anoxic. Downgradient from the tailings, total nitrogen is being removed from the groundwater system at a greater rate than the geochemically conservative anion, chloride. This removal may be due to denitrification and adsorption of ammonium onto clay and silt particles. (author)

Denitrification in groundwater at uranium mill tailings sites

Energy Technology Data Exchange (ETDEWEB)

Goering, Timothy J [Jacobs Engineering Group, Inc., Albuquerque, NM (United States); Groffman, Armando [Roy F. Weston, Inc., Albuquerque, NM (United States); Thomson, Bruce [University of New Mexico, Albuquerque, NM (United States)

1992-07-01

Nitrates are a major contaminant in groundwater at many Uranium Mill Tailings Remedial Action (UMTRA) sites. Microbial denitrification, the transformation of nitrate to nitrogen gas, may be occurring in groundwater at several UMTRA sites. Denitrification is a biologically mediated process whereby facultative anaerobes use nitrate for respiration under anaerobic conditions. Denitrifying bacteria are ubiquitous in soils, sediments, and water. Denitrification requires nitrate, organic carbon, oxygen-limiting conditions, and trace nutrients, especially phosphorus. The lack of organic carbon is the most common limiting factor for denitrification. Denitrification occurs under a limited range of temperature and pH. The uranium milling processes used at UMTRA sites provided a readily available source of carbon and nitrates for denitrifying bacteria. At the Maybell, Colorado, site, the denitrifying organisms Pseudomonas, Flavobacterium and Acinetobacter were identified in core samples of materials from beneath the tailings. In addition, microcosm experiments simulating aquifer conditions beneath the tailings pile showed an average 40 percent decrease in nitrate concentrations over 13 days. At the New Rifle, Colorado, site, aquifer conditions appear favorable for denitrification. Nitrate and organic carbon are readily available in the groundwater, and redox conditions beneath and downgradient of the tailings pile are relatively anoxic. Downgradient from the tailings, total nitrogen is being removed from the groundwater system at a greater rate than the geochemically conservative anion, chloride. This removal may be due to denitrification and adsorption of ammonium onto clay and silt particles. (author)

Plant effects on soil denitrification - a review of potential mechanisms

Science.gov (United States)

Malique, Francois; Butterbach-Bahl, Klaus; Dannenmann, Michael

2017-04-01

Denitrification is a microbial process occurring in soils, both producing and consuming the potent greenhouse gas nitrous oxide (NO), competing for nitrate with plants and hydrological leaching pathways, removing nutrients and reactive nitrogen from the biosphere, and closing the global nitrogen cycle. Despite its obvious importance, denitrification remained among the least well quantified biogeochemical processes in soils. This is due to enormous methodological difficulties involved in the direct quantification of soil microbial denitrification rates (mainly with regard to the terminal product N2) and the denitrification nitrogen gas product ratios (NO:N2O:N2), Plants may affect denitrification through a myriad of mechanisms such as e.g., competition for nitrate and water, through oxygen consumption, by regulating litter quality and changing soil pH, and via the exudation of labile carbon or secondary plant compounds involved in shaping the rhizospheric microbial community. However, plant effects on denitrification so far hardly were quantified so that the actual extent of plant control on denitrification is largely unknown. Here, we summarize the current knowledge on mechanisms how plants can affect denitrification rates and N gas product ratios in soils at temporal scales from hours to days and years. We review earlier research to quantify plant effects on denitrification as well as critically discuss the limited methods currently available to quantify plant-soil-denitrifier interactions. Finally, we provide pointers to use plants as tools to manage denitrification, e.g. to improve N use efficiency in agricultural ecosystems and to minimize soil nitrous oxide emissions.

Using natural biomass microorganisms for drinking water denitrification.

Science.gov (United States)

Costa, Darleila Damasceno; Gomes, Anderson Albino; Fernandes, Mylena; Lopes da Costa Bortoluzzi, Roseli; Magalhães, Maria de Lourdes Borba; Skoronski, Everton

2018-07-01

Among the methods that are studied to eliminate nitrate from drinking water, biological denitrification is an attractive strategy. Although several studies report the use of denitrifying bacteria for nitrate removal, they usually involve the use of sewage sludge as biomass to obtain the microbiota. In the present study, denitrifying bacteria was isolated from bamboo, and variable parameters were controlled focusing on optimal bacterial performance followed by physicochemical analysis of water adequacy. In this way, bamboo was used as a source of denitrifying microorganisms, using either Immobilized Microorganisms (IM) or Suspended Microorganisms (SM) for nitrate removal. Denitrification parameters optimization was carried out by analysis of denitrification at different pH values, temperature, nitrate concentrations, carbon sources as well as different C/N ratios. In addition, operational stability and denitrification kinetics were evaluated. Microorganisms present in the biomass responsible for denitrification were identified as Proteus mirabilis. The denitrified water was submitted to physicochemical treatment such as coagulation and flocculation to adjust to the parameters of color and turbidity to drinking water standards. Denitrification using IM occurred with 73% efficiency in the absence of an external carbon source. The use of SM provided superior denitrification efficiency using ethanol (96.46%), glucose (98.58%) or glycerol (98.5%) as carbon source. The evaluation of the operational stability allowed 12 cycles of biomass reuse using the IM and 9 cycles using the SM. After physical-chemical treatment, only SM denitrified water remained within drinking water standards parameters of color and turbidity. Copyright © 2018 Elsevier Ltd. All rights reserved.

Oxygen at nanomolar levels reversibly suppresses process rates and gene expression in anammox and denitrification in the oxygen minimum zone off northern Chile.

Science.gov (United States)

Dalsgaard, Tage; Stewart, Frank J; Thamdrup, Bo; De Brabandere, Loreto; Revsbech, Niels Peter; Ulloa, Osvaldo; Canfield, Don E; DeLong, Edward F

2014-10-28

A major percentage (20 to 40%) of global marine fixed-nitrogen loss occurs in oxygen minimum zones (OMZs). Concentrations of O2 and the sensitivity of the anaerobic N2-producing processes of anammox and denitrification determine where this loss occurs. We studied experimentally how O2 at nanomolar levels affects anammox and denitrification rates and the transcription of nitrogen cycle genes in the anoxic OMZ off Chile. Rates of anammox and denitrification were reversibly suppressed, most likely at the enzyme level. Fifty percent inhibition of N2 and N2O production by denitrification was achieved at 205 and 297 nM O2, respectively, whereas anammox was 50% inhibited at 886 nM O2. Coupled metatranscriptomic analysis revealed that transcripts encoding nitrous oxide reductase (nosZ), nitrite reductase (nirS), and nitric oxide reductase (norB) decreased in relative abundance above 200 nM O2. This O2 concentration did not suppress the transcription of other dissimilatory nitrogen cycle genes, including nitrate reductase (narG), hydrazine oxidoreductase (hzo), and nitrite reductase (nirK). However, taxonomic characterization of transcripts suggested inhibition of narG transcription in gammaproteobacteria, whereas the transcription of anammox narG, whose gene product is likely used to oxidatively replenish electrons for carbon fixation, was not inhibited. The taxonomic composition of transcripts differed among denitrification enzymes, suggesting that distinct groups of microorganisms mediate different steps of denitrification. Sulfide addition (1 µM) did not affect anammox or O2 inhibition kinetics but strongly stimulated N2O production by denitrification. These results identify new O2 thresholds for delimiting marine nitrogen loss and highlight the utility of integrating biogeochemical and metatranscriptomic analyses. The removal of fixed nitrogen via anammox and denitrification associated with low O2 concentrations in oceanic oxygen minimum zones (OMZ) is a major sink in

Denitrification: an important pathway for nitrous oxide production in tropical mangrove sediments (Goa, India).

Science.gov (United States)

Fernandes, Sheryl Oliveira; Bharathi, P A Loka; Bonin, Patricia C; Michotey, Valérie D

2010-01-01

Net nitrous oxide production and denitrification activity were measured in two mangrove ecosystems of Goa, India. The relatively pristine site Tuvem was compared to Divar, which is prone to high nutrient input. Stratified sampling at 2-cm intervals within the 0- to 10-cm depth range showed that N2O production at both the locations decreased with depth. Elevated denitrification activity at Divar resulted in maximum production of up to 1.95 nmol N2O-N g(-1) h(-1) at 2 to 4 cm, which was three times higher than at Tuvem. Detailed investigations to understand the major pathway contributing to N2O production performed at Tuvem showed that incomplete denitrification was responsible for up to 43 to 93% of N2O production. Nitrous oxide production rates closely correlated to nitrite concentration (n = 15; r = -0.47; p production. Nitrous oxide production through nitrification was below detection, affirming that denitrification is the major pathway responsible for production of the greenhouse gas. Net N2O production in these mangrove systems are comparatively higher than those reported from other natural estuarine sediments and therefore warrant mitigation measures.

Simultaneous bioreduction of nitrate and chromate using sulfur-based mixotrophic denitrification process

Energy Technology Data Exchange (ETDEWEB)

Sahinkaya, Erkan, E-mail: erkansahinkaya@yahoo.com [Istanbul Medeniyet University, Bioengineering Department, Goztepe, Istanbul (Turkey); Kilic, Adem [Harran University, Environmental Engineering Department, Osmanbey Campus, 63000 Sanliurfa (Turkey); Calimlioglu, Beste; Toker, Yasemin [Istanbul Medeniyet University, Bioengineering Department, Goztepe, Istanbul (Turkey)

2013-11-15

Highlights: • Simultaneous heterotrophic and autotrophic denitrification was stimulated. • Simultaneous bioreduction of nitrate and chromate was achieved. • Total chromium decreased <50 μg/L when the influent Cr(VI) was ≤5 mg/L. -- Abstract: This study aims at evaluating simultaneous chromate and nitrate reduction using sulfur-based mixotrophic denitrification process in a column reactor packed with elemental sulfur and activated carbon. The reactor was supplemented with methanol at C/N ratio of 1.33 or 2. Almost complete denitrification was achieved at influent NO{sub 3}{sup −}–N and Cr(VI) concentrations of 75 mg/L and 10 mg/L, respectively, and 3.7 h HRT. Maximum denitrification rate was 0.5 g NO{sub 3}{sup −}–N/(L.d) when the bioreactor was fed with 75 mg/L NO{sub 3}{sup −}–N, 150 mg/L methanol and 10 mg/L Cr(VI). The share of autotrophic denitrification was between 12% and 50% depending on HRT, C/N ratio and Cr(VI) concentration. Effluent total chromium was below 50 μg/L provided that influent Cr(VI) concentration was equal or below 5 mg/L. DGGE results showed stable microbial community throughout the operation and the presence of sulfur oxidizing denitrifying bacteria (Thiobacillus denitrificans) and Cr(VI) reducing bacteria (Exiguobacterium spp.) in the column bed.

Warming can boost denitrification disproportionately due to altered oxygen dynamics.

Directory of Open Access Journals (Sweden)

Annelies J Veraart

Full Text Available BACKGROUND: Global warming and the alteration of the global nitrogen cycle are major anthropogenic threats to the environment. Denitrification, the biological conversion of nitrate to gaseous nitrogen, removes a substantial fraction of the nitrogen from aquatic ecosystems, and can therefore help to reduce eutrophication effects. However, potential responses of denitrification to warming are poorly understood. Although several studies have reported increased denitrification rates with rising temperature, the impact of temperature on denitrification seems to vary widely between systems. METHODOLOGY/PRINCIPAL FINDINGS: We explored the effects of warming on denitrification rates using microcosm experiments, field measurements and a simple model approach. Our results suggest that a three degree temperature rise will double denitrification rates. By performing experiments at fixed oxygen concentrations as well as with oxygen concentrations varying freely with temperature, we demonstrate that this strong temperature dependence of denitrification can be explained by a systematic decrease of oxygen concentrations with rising temperature. Warming decreases oxygen concentrations due to reduced solubility, and more importantly, because respiration rates rise more steeply with temperature than photosynthesis. CONCLUSIONS/SIGNIFICANCE: Our results show that denitrification rates in aquatic ecosystems are strongly temperature dependent, and that this is amplified by the temperature dependencies of photosynthesis and respiration. Our results illustrate the broader phenomenon that coupling of temperature dependent reactions may in some situations strongly alter overall effects of temperature on ecological processes.

Enzyme Activities in Waste Water and Activated Sludge

DEFF Research Database (Denmark)

Nybroe, Ole; Jørgensen, Per Elberg; Henze, Mogens

1992-01-01

The purpose of the present study was to evaluate the potential of selected enzyme activity assays to determine microbial abundance and heterotrophic activity in waste water and activated sludge. In waste water, esterase and dehydrogenase activities were found to correlate with microbial abundance...... measured as colony forming units of heterotrophic bacteria. A panel of four enzyme activity assays, α-glucosidase, alanine-aminopeptidase, esterase and dehydrogenase were used to characterize activated sludge and anaerobic hydrolysis sludge from a pilot scale plant. The enzymatic activity profiles were...... distinctly different, suggesting that microbial populations were different, or had different physiological properties, in the two types of sludge. Enzyme activity profiles in activated sludge from four full-scale plants seemed to be highly influenced by the composition of the inlet. Addition of hydrolysed...

Linking denitrification and infiltration rates during managed groundwater recharge.

Science.gov (United States)

Schmidt, Calla M; Fisher, Andrew T; Racz, Andrew J; Lockwood, Brian S; Huertos, Marc Los

2011-11-15

We quantify relations between rates of in situ denitrification and saturated infiltration through shallow, sandy soils during managed groundwater recharge. We used thermal methods to determine time series of point-specific flow rates, and chemical and isotopic methods to assess denitrification progress. Zero order denitrification rates between 3 and 300 μmol L(-1) d(-1) were measured during infiltration. Denitrification was not detected at times and locations where the infiltration rate exceeded a threshold of 0.7 ± 0.2 m d(-1). Pore water profiles of oxygen and nitrate concentration indicated a deepening of the redoxocline at high flow rates, which reduced the thickness of the zone favorable for denitrification. Denitrification rates were positively correlated with infiltration rates below the infiltration threshold, suggesting that for a given set of sediment characteristics, there is an optimal infiltration rate for achieving maximum nitrate load reduction and improvements to water supply during managed groundwater recharge. The extent to which results from this study may be extended to other managed and natural hydrologic settings remains to be determined, but the approach taken in this study should be broadly applicable, and provides a quantitative link between shallow hydrologic and biogeochemical processes.

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

Energy Technology Data Exchange (ETDEWEB)

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

2017-09-28

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

Atmospheric nitrogen deposition influences denitrification and nitrous oxide production in lakes.

Science.gov (United States)

McCrackin, Michelle L; Elser, James J

2010-02-01

Microbially mediated denitrification is an important process that may ameliorate the effects of nitrogen (N) loading by permanently removing excess N inputs. In this study, we measured the rate of denitrification and nitrous oxide (N2O) production during denitrification in sediments from 32 Norwegian lakes at the high and low ends of a gradient of atmospheric N deposition. Denitrification and N2O production rates averaged 41.7 and 1.1 micromol N x m(-2) x h(-1), respectively, for high-deposition lakes. There was no detectable denitrification or N2O production in low-deposition lakes. Epilimnetic nitrate concentration was strongly correlated with denitrification rate (r2 = 0.67). We also measured the denitrification rate in response to experimental additions of organic carbon, nitrate, and phosphorus. Experimental nitrate additions stimulated denitrification in sediments of all lakes, regardless of N deposition level. In fact, the rate of denitrification in nitrate-amended treatments was the same magnitude for lakes in both deposition areas. These findings suggest that lake sediments possess considerable capacity to remove nitrate and that this capacity has not been saturated under conditions of chronic N loading. Further, nitrous oxide was nearly 3% of the total gaseous product during denitrification in high-deposition lakes, a fraction that is comparable to polluted marine sediments. Our findings suggest that, while lakes play an important role in N removal in the landscape, they may be a source of N2O emissions, especially in areas subject to elevated N inputs.

Monitoring N2O Production Using a cNOR Modeled Active Site

Science.gov (United States)

Griffiths, Z. G.; Hegg, E. L.; Finders, C.; Haslun, J. A.

2017-12-01

Nitrous oxide (N2O) is a potent greenhouse gas with a 100-year global warming potential 265-296 times greater than carbon dioxide (CO2). It is the leading contributor to ozone depletion and can persist in the stratosphere for approximately 114 years. Hence, understanding the sources of atmospheric N2O emissions is critical to remediating the effects of climate change. Agricultural activities are the largest contributor to N2O emissions in the U.S. with microbial nitrification and denitrification as the dominating soil processes. The enzyme cytochrome c nitric oxide reductase (cNOR) is involved in bacterial denitrification. It is often difficult to study the enzymes involved in biotic N2O production, hence, model enzymes are a useful tool. The enzyme I107EFeBMb, a sperm whale myoglobin derivative, models the active site of cNOR and was used to simulate the anaerobic reduction of NO to N2O by cNOR. Dithionite was used to induce the catalytic activity of I107EFeBMb by reducing the enzyme. However, dithionite is a strong reductant that is capable of reducing NO to N2O directly. Therefore, the dithionite-enzyme mixture was passed through a size-exclusion column to isolate the reduced enzyme. This reduced and purified enzyme was then utilized to investigate N2O production from NO. This project will provide both an enzymatic and abiotic model to study N2O production.

Substrate-Competitive Activity-Based Profiling of Ester Prodrug Activating Enzymes.

Science.gov (United States)

Xu, Hao; Majmudar, Jaimeen D; Davda, Dahvid; Ghanakota, Phani; Kim, Ki H; Carlson, Heather A; Showalter, Hollis D; Martin, Brent R; Amidon, Gordon L

2015-09-08

Understanding the mechanistic basis of prodrug delivery and activation is critical for establishing species-specific prodrug sensitivities necessary for evaluating preclinical animal models and potential drug-drug interactions. Despite significant adoption of prodrug methodologies for enhanced pharmacokinetics, functional annotation of prodrug activating enzymes is laborious and often unaddressed. Activity-based protein profiling (ABPP) describes an emerging chemoproteomic approach to assay active site occupancy within a mechanistically similar enzyme class in native proteomes. The serine hydrolase enzyme family is broadly reactive with reporter-linked fluorophosphonates, which have shown to provide a mechanism-based covalent labeling strategy to assay the activation state and active site occupancy of cellular serine amidases, esterases, and thioesterases. Here we describe a modified ABPP approach using direct substrate competition to identify activating enzymes for an ethyl ester prodrug, the influenza neuraminidase inhibitor oseltamivir. Substrate-competitive ABPP analysis identified carboxylesterase 1 (CES1) as an oseltamivir-activating enzyme in intestinal cell homogenates. Saturating concentrations of oseltamivir lead to a four-fold reduction in the observed rate constant for CES1 inactivation by fluorophosphonates. WWL50, a reported carbamate inhibitor of mouse CES1, blocked oseltamivir hydrolysis activity in human cell homogenates, confirming CES1 is the primary prodrug activating enzyme for oseltamivir in human liver and intestinal cell lines. The related carbamate inhibitor WWL79 inhibited mouse but not human CES1, providing a series of probes for analyzing prodrug activation mechanisms in different preclinical models. Overall, we present a substrate-competitive activity-based profiling approach for broadly surveying candidate prodrug hydrolyzing enzymes and outline the kinetic parameters for activating enzyme discovery, ester prodrug design, and

A simple model for simultaneous methanogenic-denitrification systems

DEFF Research Database (Denmark)

Garibay-Orijel, C.; Ahring, Birgitte Kiær; Rinderknecht-Seijas, N.

2006-01-01

We describe a useful and simple model for studies of simultaneous methanogenic-denitrification (M-D) systems. One equation predicts an inverse relationship between the percentage of electron donor channeled into dissimilatory denitrification and the loading ratio X given by grams degradable COD per...

Microbial ecology of denitrification in biological wastewater treatment.

Science.gov (United States)

Lu, Huijie; Chandran, Kartik; Stensel, David

2014-11-01

Globally, denitrification is commonly employed in biological nitrogen removal processes to enhance water quality. However, substantial knowledge gaps remain concerning the overall community structure, population dynamics and metabolism of different organic carbon sources. This systematic review provides a summary of current findings pertaining to the microbial ecology of denitrification in biological wastewater treatment processes. DNA fingerprinting-based analysis has revealed a high level of microbial diversity in denitrification reactors and highlighted the impacts of carbon sources in determining overall denitrifying community composition. Stable isotope probing, fluorescence in situ hybridization, microarrays and meta-omics further link community structure with function by identifying the functional populations and their gene regulatory patterns at the transcriptional and translational levels. This review stresses the need to integrate microbial ecology information into conventional denitrification design and operation at full-scale. Some emerging questions, from physiological mechanisms to practical solutions, for example, eliminating nitrous oxide emissions and supplementing more sustainable carbon sources than methanol, are also discussed. A combination of high-throughput approaches is next in line for thorough assessment of wastewater denitrifying community structure and function. Though denitrification is used as an example here, this synergy between microbial ecology and process engineering is applicable to other biological wastewater treatment processes. Copyright © 2014 Elsevier Ltd. All rights reserved.

Thermodynamic activity-based intrinsic enzyme kinetic sheds light on enzyme-solvent interactions.

Science.gov (United States)

Grosch, Jan-Hendrik; Wagner, David; Nistelkas, Vasilios; Spieß, Antje C

2017-01-01

The reaction medium has major impact on biocatalytic reaction systems and on their economic significance. To allow for tailored medium engineering, thermodynamic phenomena, intrinsic enzyme kinetics, and enzyme-solvent interactions have to be discriminated. To this end, enzyme reaction kinetic modeling was coupled with thermodynamic calculations based on investigations of the alcohol dehydrogenase from Lactobacillus brevis (LbADH) in monophasic water/methyl tert-butyl ether (MTBE) mixtures as a model solvent. Substrate concentrations and substrate thermodynamic activities were varied separately to identify the individual thermodynamic and kinetic effects on the enzyme activity. Microkinetic parameters based on concentration and thermodynamic activity were derived to successfully identify a positive effect of MTBE on the availability of the substrate to the enzyme, but a negative effect on the enzyme performance. In conclusion, thermodynamic activity-based kinetic modeling might be a suitable tool to initially curtail the type of enzyme-solvent interactions and thus, a powerful first step to potentially understand the phenomena that occur in nonconventional media in more detail. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:96-103, 2017. © 2016 American Institute of Chemical Engineers.

Mechanism and rate of denitrification in an agricultural watershed: Electron and mass balance along groundwater flow paths

Science.gov (United States)

Tesoriero, Anthony J.; Liebscher, Hugh; Cox, Stephen E.

2000-01-01

The rate and mechanism of nitrate removal along and between groundwater flow paths were investigated using a series of well nests screened in an unconfined sand and gravel aquifer. Intensive agricultural activity in this area has resulted in nitrate concentrations in groundwater often exceeding drinking water standards. Both the extent and rate of denitrification varied depending on the groundwater flow path. While little or no denitrification occurred in much of the upland portions of the aquifer, a gradual redox gradient is observed as aerobic upland groundwater moves deeper in the aquifer. In contrast, a sharp shallow redox gradient is observed adjacent to a third‐order stream as aerobic groundwater enters reduced sediments. An essentially complete loss of nitrate concurrent with increases in excess N2 provide evidence that denitrification occurs as groundwater enters this zone. Electron and mass balance calculations suggest that iron sulfide (e.g., pyrite) oxidation is the primary source of electrons for denitrification. Denitrification rate estimates were based on mass balance calculations using nitrate and excess N2 coupled with groundwater travel times. Travel times were determined using a groundwater flow model and were constrained by chlorofluorocarbon‐based age dates. Denitrification rates were found to vary considerably between the two areas where denitrification occurs. Denitrification rates in the deep, upland portions of the aquifer were found to range from from 1.0 to 2.7 mM of N per year. Potential denitrification rates in groundwater adjacent to the stream may be much faster, with rates up to 140 mM per year based on an in situ experiment conducted in this zone.

Relative importance of plant uptake and plant associated denitrification for removal of nitrogen from mine drainage in sub-arctic wetlands.

Science.gov (United States)

Hallin, Sara; Hellman, Maria; Choudhury, Maidul I; Ecke, Frauke

2015-11-15

Reactive nitrogen (N) species released from undetonated ammonium-nitrate based explosives used in mining or other blasting operations are an emerging environmental problem. Wetlands are frequently used to treat N-contaminated water in temperate climate, but knowledge on plant-microbial interactions and treatment potential in sub-arctic wetlands is limited. Here, we compare the relative importance of plant uptake and denitrification among five plant species commonly occurring in sub-arctic wetlands for removal of N in nitrate-rich mine drainage in northern Sweden. Nitrogen uptake and plant associated potential denitrification activity and genetic potential for denitrification based on quantitative PCR of the denitrification genes nirS, nirK, nosZI and nosZII were determined in plants growing both in situ and cultivated in a growth chamber. The growth chamber and in situ studies generated similar results, suggesting high relevance and applicability of results from growth chamber experiments. We identified denitrification as the dominating pathway for N-removal and abundances of denitrification genes were strong indicators of plant associated denitrification activity. The magnitude and direction of the effect differed among the plant species, with the aquatic moss Drepanocladus fluitans showing exceptionally high ratios between denitrification and uptake rates, compared to the other species. However, to acquire realistic estimates of N-removal potential of specific wetlands and their associated plant species, the total plant biomass needs to be considered. The species-specific plant N-uptake and abundance of denitrification genes on the root or plant surfaces were affected by the presence of other plant species, which show that both multi- and inter-trophic interactions are occurring. Future studies on N-removal potential of wetland plant species should consider how to best exploit these interactions in sub-arctic wetlands. Copyright © 2015 Elsevier Ltd. All rights

Visualization of enzyme activities inside earthworm pores

Science.gov (United States)

Hoang, Duyen; Razavi, Bahar S.

2015-04-01

In extremely dynamic microhabitats as bio-pores made by earthworm, the in situ enzyme activities are assumed as a footprint of complex biotic interactions. Our study focused on the effect of earthworm on the enzyme activities inside bio-pores and visualizing the differences between bio-pores and earthworm-free soil by zymography technique (Spohn and Kuzyakov, 2013). For the first time, we aimed at quantitative imaging of enzyme activities in bio-pores. Lumbricus terrestris L. was placed into transparent box (15×20×15cm). After two weeks when bio-pore systems were formed by earthworms, we visualized in situ enzyme activities of five hydrolytic enzymes (β-glucosidase, cellobiohydrolase, chitinase, xylanase, leucine-aminopeptidase, and phosphatase. Zymography showed higher activity of β-glucosidase, chitinase, xylanase and phosphatase in biopores comparing to bulk soil. However, the differences in activity of cellobiohydrolase and leucine aminopeptidase between bio-pore and bulk soil were less pronounced. This demonstrated an applicability of zymography approach to monitor and to distinguish the in situ activity of hydrolytic enzymes in soil biopores.

Kinetics of nitrous oxide production by denitrification in municipal solid waste.

Science.gov (United States)

Wu, Chuanfu; Shimaoka, Takayuki; Nakayama, Hirofumi; Komiya, Teppei

2015-04-01

As one of the Nitrous Oxide (N2O) production pathways, denitrification plays an important role in regulating the emission of N2O into the atmosphere. In this study, the influences of different substrate concentrations and transient conditions on the denitrification rate and N2O-reducing activities were investigated. Results revealed that N2O production rates (i.e. denitrification rates) were stimulated by increased total organic carbon (TOC) concentration, while it was restrained under high oxygen concentrations. Moreover, the impact of nitrate concentrations on N2O production rates depended on the TOC/NO3--N ratios. All the N2O production rate data fitted well to a multiplicative Monod equation, with terms describing the influence of TOC and nitrate concentrations, and an Arrhenius-type equation. Furthermore, results demonstrated that high temperatures minimized the N2O-reducing activities in aged municipal solid waste, resulting in an accumulation of N2O. On the other hand, a transient condition caused by changing O2 concentrations may strongly influence the N2O production rates and N2O-reducing activities in solid waste. Finally, based on the results, we believe that a landfill aeration strategy properly designed to prevent rising temperatures and to cycle air injection is the key to reducing emissions of N2O during remediation of old landfills by means of in situ aeration. Copyright © 2015 Elsevier Ltd. All rights reserved.

County-Scale Spatial Distribution of Soil Enzyme Activities and Enzyme Activity Indices in Agricultural Land: Implications for Soil Quality Assessment

Directory of Open Access Journals (Sweden)

Xiangping Tan

2014-01-01

Full Text Available Here the spatial distribution of soil enzymatic properties in agricultural land was evaluated on a county-wide (567 km2 scale in Changwu, Shaanxi Province, China. The spatial variations in activities of five hydrolytic enzymes were examined using geostatistical methods. The relationships between soil enzyme activities and other soil properties were evaluated using both an integrated total enzyme activity index (TEI and the geometric mean of enzyme activities (GME. At the county scale, soil invertase, phosphatase, and catalase activities were moderately spatially correlated, whereas urease and dehydrogenase activities were weakly spatially correlated. Correlation analysis showed that both TEI and GME were better correlated with selected soil physicochemical properties than single enzyme activities. Multivariate regression analysis showed that soil OM content had the strongest positive effect while soil pH had a negative effect on the two enzyme activity indices. In addition, total phosphorous content had a positive effect on TEI and GME in orchard soils, whereas alkali-hydrolyzable nitrogen and available potassium contents, respectively, had negative and positive effects on these two enzyme indices in cropland soils. The results indicate that land use changes strongly affect soil enzyme activities in agricultural land, where TEI provides a sensitive biological indicator for soil quality.

Use of bioreactor landfill for nitrogen removal to enhance methane production through ex situ simultaneous nitrification-denitrification and in situ denitrification.

Science.gov (United States)

Sun, Xiaojie; Zhang, Hongxia; Cheng, Zhaowen

2017-08-01

High concentrations of nitrate-nitrogen (NO 3 - -N) derived from ex situ nitrification phase can inhibit methane production during ex situ nitrification and in situ denitrification bioreactor landfill. A combined process comprised of ex situ simultaneous nitrification-denitrification (SND) in an aged refuse bioreactor (ARB) and in situ denitrification in a fresh refuse bioreactor (FRB) was conducted to reduce the negative effect of high concentrationsof NO 3 - -N. Ex situ SND can be achieved because NO 3 - -N concentration can be reduced and the removal rate of ammonium-nitrogen (NH 4 + -N) remains largely unchanged when the ventilation rate of ARB-A2 is controlled. The average NO 3 - -N concentrations of effluent were 470mg/L in ex situ nitrification ARB-A1 and 186mg/L in ex situ SND ARB-A2. The average NH 4 + -N removal rates of ARB-A1 and ARB-A2 were 98% and 94%, respectively. Based on the experimental data from week 4 to week 30, it is predicted that NH 4 + -N concentration in FRB-F1 of the ex situ nitrification and in situ denitrification process would reach 25mg/L after 63weeks, and about 40weeks for the FRB-F2 of ex situ SND and in situ denitrification process . Ex situ SND and in situ denitrification process can improve themethane production of FRB-F2. The lag phase time of methane production for the FRB-F2 was 11weeks. This phase was significantly shorter than the 15-week phases of FRB-F1 in ex situ nitrification and in situ denitrification process. A seven-week stabilizationphase was required to increase methane content from 5% to 50% for FRB-F2. Methane content in FRB-F1 did not reach 50% but reached the 45% peak after 20weeks. Copyright © 2017 Elsevier Ltd. All rights reserved.

Denitrification and inference of nitrogen sources in the karstic Floridan Aquifer

Directory of Open Access Journals (Sweden)

J. B. Heffernan

2012-05-01

Full Text Available Aquifer denitrification is among the most poorly constrained fluxes in global and regional nitrogen budgets. The few direct measurements of denitrification in groundwaters provide limited information about its spatial and temporal variability, particularly at the scale of whole aquifers. Uncertainty in estimates of denitrification may also lead to underestimates of its effect on isotopic signatures of inorganic N, and thereby confound the inference of N source from these data. In this study, our objectives are to quantify the magnitude and variability of denitrification in the Upper Floridan Aquifer (UFA and evaluate its effect on N isotopic signatures at the regional scale. Using dual noble gas tracers (Ne, Ar to generate physical predictions of N₂ gas concentrations for 112 observations from 61 UFA springs, we show that excess (i.e. denitrification-derived N₂ is highly variable in space and inversely correlated with dissolved oxygen (O₂. Negative relationships between O₂ and δ¹⁵N_NO3 across a larger dataset of 113 springs, well-constrained isotopic fractionation coefficients, and strong ¹⁵N:¹⁸O covariation further support inferences of denitrification in this uniquely organic-matter-poor system. Despite relatively low average rates, denitrification accounted for 32 % of estimated aquifer N inputs across all sampled UFA springs. Back-calculations of source δ¹⁵N_NO3 based on denitrification progression suggest that isotopically-enriched nitrate (NO₃^– in many springs of the UFA reflects groundwater denitrification rather than urban- or animal-derived inputs.

Nitrate levels modulate denitrification activity in tropical mangrove sediments (Goa, India)

Digital Repository Service at National Institute of Oceanography (India)

Fernandes, S.O.; LokaBharathi, P.A.

.M., & Bundrick, C.M. (1998). Potential sediment denitrification rates in estuaries of Northern Gulf of Mexico. Journal of environmental quality, 27(4), 859-868. Gruber, N., & Sarmiento, J.L. (1997). Global. Biogeochemical Cycles, 11, 235-266. Hahndel, R... of inorganic nitrogen in mangrove sediments (Terminos Lagoon, Mexico). Limnology and Oceanography, 41, 284-296. Sardessai, S. (1993). Dissolved, particulate and sedimentary humic acids in the mangroves and estuarine ecosystem of Goa, west coast of India...

Lysosomal enzyme activation in irradiated mammary tumors

International Nuclear Information System (INIS)

Clarke, C.; Wills, E.D.

1976-01-01

Lysosomal enzyme activity of C3H mouse mammary tumors was measured quantitatively by a histochemical method. Following whole-body doses of 3600 rad or less no changes were observed in the lysosomal enzyme activity for 12 hr after the irradiation, but very large increases in acid phosphatase and β-naphthylamidase activity were, however, observed 24 hr after irradiation. Significant increases in enzyme activity were detected 72 hr after a dose of 300 rad and the increases of enzyme activity were dose dependent over the range 300 to 900 rad. Testosterone (80 mg/kg) injected into mice 2 hr before irradiation (850 rad) caused a significant increase of lysosomal enzyme activity over and above that of the same dose of irradiation alone. If the tumor-bearing mice were given 95 percent oxygen/5 percent carbon dioxide to breathe for 8 min before irradiation the effect of 850 rad on lysosomal acid phosphatase was increased to 160 percent/that of the irradiation given alone. Activitation of lysosomal enzymes in mammary tumors is an important primary or secondary consequence of radiation

Impact of chloride on denitrification potential in roadside wetlands

International Nuclear Information System (INIS)

Lancaster, Nakita A.; Bushey, Joseph T.; Tobias, Craig R.; Song, Bongkeun; Vadas, Timothy M.

2016-01-01

Developed landscapes are exposed to changes in hydrology and water chemistry that limit their ability to mitigate detrimental impacts to coastal water bodies, particularly those that result from stormwater runoff. The elevated level of impervious cover increases not only runoff but also contaminant loading of nutrients, metals, and road salt used for deicing to water bodies. Here we investigate the impact that road salt has on denitrification in roadside environments. Sediments were collected from a series of forested and roadside wetlands and acclimated with a range of Cl − concentrations from 0 to 5000 mg L −1 for 96 h. Denitrification rates were measured by the isotope pairing technique using 15 N–NO 3 − , while denitrifying community structures were compared using terminal restriction fragment length polymorphism (T-RFLP) of nitrous oxide reductase genes (nosZ). Chloride significantly (p < 0.05) inhibited denitrification in forested wetlands at a Cl − dosage of 2500 or 5000 mg L −1 , but the decrease in denitrification rates was less and not significant for the roadside wetlands historically exposed to elevated concentrations of Cl − . The difference could not be attributed to other significant changes in conditions, such as DOC concentrations, N species concentrations, or pH levels. Denitrifying communities, as measured by T-RFs of the nosZ gene, in the roadside wetlands with elevated concentration of Cl − were distinctly different and more diverse compared to forested wetlands, and also different in roadside wetlands after 96 h exposures to Cl − . The shifts in denitrifying communities seem to minimize the decrease in denitrification rates in the wetlands previously exposed to Cl. As development results in more Cl − use and exposure to a broad range of natural or manmade wetland structures, an understanding of the seasonal effect of Cl on denitrification processes in these systems would aid in design or mitigation of the effects on

Epigenetics of dominance for enzyme activity

Indian Academy of Sciences (India)

Unknown

dimer over a wide range of H+ concentrations accounts for the epigenetics of dominance for enzyme activity. [Trehan K S ... The present study has been carried on acid phosphatase .... enzyme activity over mid parent value (table 3, col. 13),.

SNCR method of flue gas denitrification

Energy Technology Data Exchange (ETDEWEB)

Kuropka, J. [Politechniki Wroclawskiej, Wroclaw (Poland). Instytut Inzynierii Ochrony Srodowiska

1998-12-31

Current achievements in experiments on selective non-catalytic reduction of nitrogen oxides from flue gases were presented. Some basic parameters of denitrification process (temperature of reaction, contact time, molar ratio of agents, additions to reacting substances) which influence the rate of nitrogen oxides emission from flue gases were analysed. On the basis of conducted experiments with calcium hydroxide and urea or calcium carbonate and urea on full-scale FGD installation on WP-120 boiler it was found that SNCR method can be applied to simultaneous denitrification and desulfurisation of flue gases. 27 refs., 10 figs.

Controls on denitrification potential in nitrate-rich waterways and riparian zones of an irrigated agricultural setting.

Science.gov (United States)

Webster, Alex J; Groffman, Peter M; Cadenasso, Mary L

2018-02-21

Denitrification, the microbial conversion of NO 3 - to N gases, is an important process contributing to whether lotic and riparian ecosystems act as sinks for excess NO 3 - from agricultural activities. Though agricultural waterways and riparian zones have been a focus of denitrification research for decades, almost none of this research has occurred in the irrigated agricultural settings of arid and semi-arid climates. In this study, we conducted a broad survey of denitrification potential in riparian soils and channel sediment from 79 waterway reaches in the irrigated agricultural landscape of California's Central Valley. With this approach, we sought to capture the wide range of variation that arose from diverse waterway management and fluctuating flow conditions, and use this variation to identify promising management interventions. We explored associations of denitrification potentials with surface water NO 3 - -N, organic matter, flow conditions, vegetation cover, near-channel riparian bank slope, and channel geomorphic features using generalized linear mixed models. We found strong associations of sediment denitrification potentials with reach flow conditions, which we hypothesize was the result of variation in microbial communities' tolerance to dry-wet cycles. Denitrification potentials in riparian soils, in contrast, did not appear affected by flow conditions, but instead were associated with organic matter, vegetation cover, and bank slope in the riparian zone. These results suggest a strong need for further work on how denitrification responds to varying flow conditions and dry-wet cycles in non-perennial lotic ecosystems. Our findings also demonstrate that denitrifier communities respond to key features of waterway management, which can therefore be leveraged to control denitrification through a variety of management actions. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Heterotrophic Anodic Denitrification in Microbial Fuel Cells

Directory of Open Access Journals (Sweden)

Jakub Drewnowski

2016-06-01

Full Text Available Nowadays, pollution caused by energy production systems is a major environmental concern. Therefore, the development of sustainable energy sources is required. Amongst others, the microbial fuel cell (MFC seems to be a possible solution because it can produce clean energy at the same time that waste is stabilized. Unfortunately, mainly due to industrial discharges, the wastes could contain nitrates, or nitrates precursors such ammonia, which could lead to lower performance in terms of electricity production. In this work, the feasibility of coupling anodic denitrification process with electricity production in MFC and the effect of the nitrates over the MFC performance were studied. During the experiments, it was observed that the culture developed in the anodic chamber of the MFC presented a significant amount of denitrificative microorganisms. The MFC developed was able to denitrify up to 4 ppm, without affecting the current density exerted, of about 1 mA/cm2. Regarding the denitrification process, it must be highlighted that the maximum denitrification rate achieved with the culture was about 60 mg·NO3−·L−1·h−1. Based on these results, it can be stated that it is possible to remove nitrates and to produce energy, without negatively affecting the electrical performance, when the nitrate concentration is low.

Spatial distribution of enzyme activities in the rhizosphere

Science.gov (United States)

Razavi, Bahar S.; Zarebanadkouki, Mohsen; Blagodatskaya, Evgenia; Kuzyakov, Yakov

2015-04-01

The rhizosphere, the tiny zone of soil surrounding roots, certainly represents one of the most dynamic habitat and interfaces on Earth. Activities of enzymes produced by both plant roots and microbes are the primary biological drivers of organic matter decomposition and nutrient cycling. That is why there is an urgent need in spatially explicit methods for the determination of the rhizosphere extension and enzyme distribution. Recently, zymography as a new technique based on diffusion of enzymes through the 1 mm gel plate for analysis has been introduced (Spohn & Kuzyakov, 2013). We developed the zymography technique to visualize the enzyme activities with a higher spatial resolution. For the first time, we aimed at quantitative imaging of enzyme activities as a function of distance from the root tip and the root surface in the soil. We visualized the two dimensional distribution of the activity of three enzymes: β-glucosidase, phosphatase and leucine amino peptidase in the rhizosphere of maize using fluorogenically labelled substrates. Spatial-resolution of fluorescent images was improved by direct application of a substrate saturated membrane to the soil-root system. The newly-developed direct zymography visualized heterogeneity of enzyme activities along the roots. The activity of all enzymes was the highest at the apical parts of individual roots. Across the roots, the enzyme activities were higher at immediate vicinity of the roots (1.5 mm) and gradually decreased towards the bulk soil. Spatial patterns of enzyme activities as a function of distance from the root surface were enzyme specific, with highest extension for phosphatase. We conclude that improved zymography is promising in situ technique to analyze, visualize and quantify spatial distribution of enzyme activities in the rhizosphere hotspots. References Spohn, M., Kuzyakov, Y., 2013. Phosphorus mineralization can be driven by microbial need for carbon. Soil Biology & Biochemistry 61: 69-75

The kinetics of denitrification in permeable sediments

DEFF Research Database (Denmark)

Evrard, Victor; Glud, Ronnie N.; Cook, Perran L. M.

2013-01-01

Permeable sediments comprise the majority of shelf sediments, yet the rates of denitrification remain highly uncertain in these environments. Computational models are increasingly being used to understand the dynamics of denitrification in permeable sediments, which are complex environments...... on sediments taken from six shallow coastal sites in Port Phillip Bay, Victoria, Australia. The results showed that denitrification commenced rapidly (within 30 min) after the onset of anoxia and the kinetics could be well described by Michaelis-Menten kinetics with half saturation constants (apparent K...... in cohesive sediments despite organic carbon contents one order of magnitude lower for the sediments studied here. The ratio of sediment O-2 consumption to V-max was in the range of 0.02-0.09, and was on average much lower than the theoretical ratio of 0.8. As a consequence, models implemented...

Autoxidation and acetylene-accelerated oxidation of NO in a 2-phase system; implications for the expression of denitrification in ex situ experiments

Science.gov (United States)

Nadeem, Shahid; Dörsch, Peter; Bakken, Lars

2013-04-01

Denitrification allows microorganisms to sustain respiration under anoxic conditions. The typical niche for denitrification is an environment with fluctuating oxygen concentrations such as soils and borders between anoxic and oxic zones of biofilms and sediments. In such environments, the organisms need adequate regulation of denitrification in response to changing oxygen availability to tackle both oxic and anoxic spells. The regulation of denitrification in soils has environmental implications, since it affects the proportions of N2, N2O and NO emitted to the atmosphere. The expression of denitrification enzymes is regulated by a complex regulatory network involving one or several positive feedback loops via the intermediate nitrogen oxides. Nitric oxide (NO) is known to induce denitrification in model organisms, but the quantitative effect of NO and its concentration dependency has not been assessed for denitrification in soils. NO is chemically unstable in the presence of oxygen due to autoxidation, and the oxidation of NO is accelerated by acetylene (C2H2) which is commonly used as an inhibitor of N2O reductase in denitrification studies. As a first step to a better understanding of NO's role in soil denitrification, we investigated NO oxidation kinetics for a closed "two phase" system (i.e. liquid phase + headspace) typically used for denitrification experiments with soil slurries, with and without acetylene present. Models were developed to adequately predict autoxidation and acetylene-accelerated oxidation. The minimum oxygen concentration in the headspace ([O2]min, mL L-1) for acetylene-accelerated NO oxidation was found to increase linearly with the NO concentration ([NO], mL L-1); [O2]min= 0.192 + [NO]*0.1 (r2=0.978). The models for NO oxidation were then used to assess NO-oxidation rates in denitrification experiments with batches of bacterial cells extracted from soil. The batches were exposed to low initial oxygen concentrations in gas tight serum

Intergenomic comparisons highlight modularity of the denitrification pathway and underpin the importance of community structure for N2O emissions.

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Daniel R H Graf

Full Text Available Nitrous oxide (N2O is a potent greenhouse gas and the predominant ozone depleting substance. The only enzyme known to reduce N2O is the nitrous oxide reductase, encoded by the nosZ gene, which is present among bacteria and archaea capable of either complete denitrification or only N2O reduction to di-nitrogen gas. To determine whether the occurrence of nosZ, being a proxy for the trait N2O reduction, differed among taxonomic groups, preferred habitats or organisms having either NirK or NirS nitrite reductases encoded by the nirK and nirS genes, respectively, 652 microbial genomes across 18 phyla were compared. Furthermore, the association of different co-occurrence patterns with enzymes reducing nitric oxide to N2O encoded by nor genes was examined. We observed that co-occurrence patterns of denitrification genes were not randomly distributed across taxa, as specific patterns were found to be more dominant or absent than expected within different taxonomic groups. The nosZ gene had a significantly higher frequency of co-occurrence with nirS than with nirK and the presence or absence of a nor gene largely explained this pattern, as nirS almost always co-occurred with nor. This suggests that nirS type denitrifiers are more likely to be capable of complete denitrification and thus contribute less to N2O emissions than nirK type denitrifiers under favorable environmental conditions. Comparative phylogenetic analysis indicated a greater degree of shared evolutionary history between nosZ and nirS. However 30% of the organisms with nosZ did not possess either nir gene, with several of these also lacking nor, suggesting a potentially important role in N2O reduction. Co-occurrence patterns were also non-randomly distributed amongst preferred habitat categories, with several habitats showing significant differences in the frequencies of nirS and nirK type denitrifiers. These results demonstrate that the denitrification pathway is highly modular, thus

Denitrification in agriculturally impacted streams: seasonal changes in structure and function of the bacterial community.

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Erin Manis

Full Text Available Denitrifiers remove fixed nitrogen from aquatic environments and hydrologic conditions are one potential driver of denitrification rate and denitrifier community composition. In this study, two agriculturally impacted streams in the Sugar Creek watershed in Indiana, USA with different hydrologic regimes were examined; one stream is seasonally ephemeral because of its source (tile drainage, whereas the other stream has permanent flow. Additionally, a simulated flooding experiment was performed on the riparian benches of the ephemeral stream during a dry period. Denitrification activity was assayed using the chloramphenicol amended acetylene block method and bacterial communities were examined based on quantitative PCR and terminal restriction length polymorphisms of the nitrous oxide reductase (nosZ and 16S rRNA genes. In the stream channel, hydrology had a substantial impact on denitrification rates, likely by significantly lowering water potential in sediments. Clear patterns in denitrification rates were observed among pre-drying, dry, and post-drying dates; however, a less clear scenario was apparent when analyzing bacterial community structure suggesting that denitrifier community structure and denitrification rate were not strongly coupled. This implies that the nature of the response to short-term hydrologic changes was physiological rather than increases in abundance of denitrifiers or changes in composition of the denitrifier community. Flooding of riparian bench soils had a short-term, transient effect on denitrification rate. Our results imply that brief flooding of riparian zones is unlikely to contribute substantially to removal of nitrate (NO3- and that seasonal drying of stream channels has a negative impact on NO3- removal, particularly because of the time lag required for denitrification to rebound. This time lag is presumably attributable to the time required for the denitrifiers to respond physiologically rather than a change

Studies on benthic denitrification in the Chwaka Bay mangrove ...

African Journals Online (AJOL)

Spatial variations in denitrification rates were due to variations in concentration levels of organic matter and possibly to disproportionate competition for inorganic nitrogen between denitrifiers and benthic autotrophs among sites. There were no seasonal differences in denitrification rates. Results from the present study ...

Enzymes and Enzyme Activity Encoded by Nonenveloped Viruses.

Science.gov (United States)

Azad, Kimi; Banerjee, Manidipa; Johnson, John E

2017-09-29

Viruses are obligate intracellular parasites that rely on host cell machineries for their replication and survival. Although viruses tend to make optimal use of the host cell protein repertoire, they need to encode essential enzymatic or effector functions that may not be available or accessible in the host cellular milieu. The enzymes encoded by nonenveloped viruses-a group of viruses that lack any lipid coating or envelope-play vital roles in all the stages of the viral life cycle. This review summarizes the structural, biochemical, and mechanistic information available for several classes of enzymes and autocatalytic activity encoded by nonenveloped viruses. Advances in research and development of antiviral inhibitors targeting specific viral enzymes are also highlighted.

Enzyme activity assay of glycoprotein enzymes based on a boronate affinity molecularly imprinted 96-well microplate.

Science.gov (United States)

Bi, Xiaodong; Liu, Zhen

2014-12-16

Enzyme activity assay is an important method in clinical diagnostics. However, conventional enzyme activity assay suffers from apparent interference from the sample matrix. Herein, we present a new format of enzyme activity assay that can effectively eliminate the effects of the sample matrix. The key is a 96-well microplate modified with molecularly imprinted polymer (MIP) prepared according to a newly proposed method called boronate affinity-based oriented surface imprinting. Alkaline phosphatase (ALP), a glycoprotein enzyme that has been routinely used as an indicator for several diseases in clinical tests, was taken as a representative target enzyme. The prepared MIP exhibited strong affinity toward the template enzyme (with a dissociation constant of 10(-10) M) as well as superb tolerance for interference. Thus, the enzyme molecules in a complicated sample matrix could be specifically captured and cleaned up for enzyme activity assay, which eliminated the interference from the sample matrix. On the other hand, because the boronate affinity MIP could well retain the enzymatic activity of glycoprotein enzymes, the enzyme captured by the MIP was directly used for activity assay. Thus, additional assay time and possible enzyme or activity loss due to an enzyme release step required by other methods were avoided. Assay of ALP in human serum was successfully demonstrated, suggesting a promising prospect of the proposed method in real-world applications.

The ultrasound technology for modifying enzyme activity

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Meliza Lindsay

2016-06-01

Full Text Available Enzymes are protein complexes compounds widely studied and used due to their ability to catalyze reactions. The food processing mainly aims the inactivation of enzymes due to various undesirable effects. However, there are many processes that can be optimized by its catalytic activity. In this context, different technologies have been applied both to inactivate or to improve the enzymes efficiency. The Ultrasound technology emerges as an alternative mainly applied to achieve the enzyme inactivation. On the contrary, very few investigations show the ability of this technology under certain conditions to achieve the opposite effect (i.e. increase the catalytic activity of enzymes. The objective of this study was to correlate the ultrasonic energy delivered to the sample (J/mL with the residual enzymatic activity and explain the possible mechanisms which results in the enzymatic activation/inactivation complex behavior. The activity of POD in coconut water was evaluated as a model. The enzymatic activity initially increased, followed by reduction with a trend to enzyme inactivation. This complex behavior is directly related to the applied ultrasonic energy and their direct mechanical effects on the product, as well as the effect in the enzymatic infinite intermediate states and its structural conformation changes. The obtained results are useful for both academic and industrial perspectives.

Lignin-degrading enzyme activities.

Science.gov (United States)

Chen, Yi-ru; Sarkanen, Simo; Wang, Yun-Yan

2012-01-01

Over the past three decades, the activities of four kinds of enzyme have been purported to furnish the mechanistic foundations for macromolecular lignin depolymerization in decaying plant cell walls. The pertinent fungal enzymes comprise lignin peroxidase (with a relatively high redox potential), manganese peroxidase, an alkyl aryl etherase, and laccase. The peroxidases and laccase, but not the etherase, are expressed extracellularly by white-rot fungi. A number of these microorganisms exhibit a marked preference toward lignin in their degradation of lignocellulose. Interestingly, some white-rot fungi secrete both kinds of peroxidase but no laccase, while others that are equally effective express extracellular laccase activity but no peroxidases. Actually, none of these enzymes has been reported to possess significant depolymerase activity toward macromolecular lignin substrates that are derived with little chemical modification from the native biopolymer. Here, the assays commonly employed for monitoring the traditional fungal peroxidases, alkyl aryl etherase, and laccase are described in their respective contexts. A soluble native polymeric substrate that can be isolated directly from a conventional milled-wood lignin preparation is characterized in relation to its utility in next-generation lignin-depolymerase assays.

The ultrasound technology for modifying enzyme activity

Directory of Open Access Journals (Sweden)

Meliza Lindsay Rojas

2016-01-01

Full Text Available Enzymes are protein complexes compounds widely studied and used due to their ability to catalyze reactions. The food processing mainly a ims the inactivation of enzymes due to various undesirable effects. However, there are many processes that can be optimized by its catalytic activity. In this context, different technologies have been applied both to inactivate or to improve the enzymes ef ficiency. The Ultrasound technology emerges as an alternative mainly applied to achieve the enzyme inactivation. On the contrary, very few investigations show the ability of this technology under certain conditions to achieve the opposite effect (i.e. increase the catalytic activity of enzymes. The objective of this study was to correlate the ultrasonic energy delivered to the sample (J/mL with the residual enzymatic activity and explain the possible mechanisms which results in the enzymatic activation/in activation complex behavior. The activity of POD in coconut water was evaluated as a model. The enzymatic activity initially increased, followed by reduction with a trend to enzyme inactivation. This complex behavior is directly related to the applied ultr asonic energy and their direct mechanical effects on the product, as well as the effect in the enzymatic infinite intermediate states and its structural conformation changes. The obtained results are useful for both academic and industrial perspectives.

Biological nitrate removal from water and wastewater by solid-phase denitrification process.

Science.gov (United States)

Wang, Jianlong; Chu, Libing

2016-11-01

Nitrate pollution in receiving waters has become a serious issue worldwide. Solid-phase denitrification process is an emerging technology, which has received increasing attention in recent years. It uses biodegradable polymers as both the carbon source and biofilm carrier for denitrifying microorganisms. A vast array of natural and synthetic biopolymers, including woodchips, sawdust, straw, cotton, maize cobs, seaweed, bark, polyhydroxyalkanoate (PHA), polycaprolactone (PCL), polybutylene succinate (PBS) and polylactic acid (PLA), have been widely used for denitrification due to their good performance, low cost and large available quantities. This paper presents an overview on the application of solid-phase denitrification in nitrate removal from drinking water, groundwater, aquaculture wastewater, the secondary effluent and wastewater with low C/N ratio. The types of solid carbon source, the influencing factors, the microbial community of biofilm attached on the biodegradable carriers, the potential adverse effect, and the cost of denitrification process are introduced and evaluated. Woodchips and polycaprolactone are the popular and competitive natural plant-like and synthetic biodegradable polymers used for denitrification, respectively. Most of the denitrifiers reported in solid-phase denitrification affiliated to the family Comamonadaceae in the class Betaproteobacteria. The members of genera Diaphorobacter, Acidovorax and Simplicispira were mostly reported. In future study, more attention should be paid to the simultaneous removal of nitrate and toxic organic contaminants such as pesticide and PPCPs by solid-phase denitrification, to the elucidation of the metabolic and regulatory relationship between decomposition of solid carbon source and denitrification, and to the post-treatment of the municipal secondary effluent. Solid-phase denitrification process is a promising technology for the removal of nitrate from water and wastewater. Copyright © 2016

Denitrification pathways and rates in the sandy sediments of the Georgia continental shelf, USA

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Ingall Ellery

2005-02-01

Full Text Available Denitrification in continental shelf sediments has been estimated to be a significant sink of oceanic fixed nitrogen (N. The significance and mechanisms of denitrification in organic-poor sands, which comprise 70% of continental shelf sediments, are not well known. Core incubations and isotope tracer techniques were employed to determine processes and rates of denitrification in the coarse-grained, sandy sediments of the Georgia continental shelf. In these sediments, heterotrophic denitrification was the dominant process for fixed N removal. Processes such as coupled nitrification-denitrification, anammox (anaerobic ammonium oxidation, and oxygen-limited autotrophic nitrification-denitrification were not evident over the 24 and 48 h time scale of the incubation experiments. Heterotrophic denitrification processes produce 22.8–34.1 μmole N m-2 d-1 of N2 in these coarse-grained sediments. These denitrification rates are approximately two orders of magnitude lower than rates determined in fine-grained shelf sediments. These lower rates may help reconcile unbalanced marine N budgets which calculate global N losses exceeding N inputs.

Enzyme with rhamnogalacturonase activity.

NARCIS (Netherlands)

Kofod, L.V.; Andersen, L.N.; Dalboge, H.; Kauppinen, M.S.; Christgau, S.; Heldt-Hansen, H.P.; Christophersen, C.; Nielsen, P.M.; Voragen, A.G.J.; Schols, H.A.

1998-01-01

An enzyme exhibiting rhamnogalacturonase activity, capable of cleaving a rhamnogalacturonan backbone in such a manner that galacturonic acids are left as the non-reducing ends, and which exhibits activity on hairy regions from a soy bean material and/or on saponified hairy regions from a sugar beet

Removal of highly elevated nitrate from drinking water by pH-heterogenized heterotrophic denitrification facilitated with ferrous sulfide-based autotrophic denitrification.

Science.gov (United States)

Huang, Bin; Chi, Guangyu; Chen, Xin; Shi, Yi

2011-11-01

The performance of acetic acid-supported pH-heterogenized heterotrophic denitrification (HD) facilitated with ferrous sulfide-based autotrophic denitrification (AD) was investigated in upflow activated carbon-packed column reactors for reliable removal of highly elevated nitrate (42 mg NO(3)-Nl(-1)) in drinking water. The use of acetic acid as substrate provided sufficient internal carbon dioxide to completely eliminate the need of external pH adjustment for HD, but simultaneously created vertically heterogenized pH varying from 4.8 to 7.8 in the HD reactor. After 5-week acclimation, the HD reactor developed a moderate nitrate removal capacity with about one third of nitrate removal occurring in the acidic zone (pH 4.8-6.2). To increase the treatment reliability, acetic acid-supported HD was operated under 10% carbon limitation to remove >85% of nitrate, and ferrous sulfide-based AD was supplementally operated to remove residual nitrate and formed nitrite without excess of soluble organic carbon, nitrite or sulfate in the final effluent. Copyright © 2011 Elsevier Ltd. All rights reserved.

Distributions, abundances and activities of microbes associated with the nitrogen cycle in riparian and stream sediments of a river tributary.

Science.gov (United States)

Kim, Haryun; Bae, Hee-Sung; Reddy, K Ramesh; Ogram, Andrew

2016-12-01

River tributaries are ecologically important environments that function as sinks of inorganic nitrogen. To gain greater insight into the nitrogen cycle (N-cycle) in these environments, the distributions and activities of microbial populations involved in the N-cycle were studied in riparian and stream sediments of the Santa Fe River (SFR) tributaries located in northern Florida, USA. Riparian sediments were characterized by much higher organic matter content, and extracellular enzyme activities, including cellobiohydrolase, β-d-glucosidase, and phenol oxidase than stream sediments. Compared with stream sediments, riparian sediments exhibited significantly higher activities of nitrification, denitrification, dissimilatory nitrate reduction to ammonia (DNRA) and anaerobic ammonia oxidation; correspondingly, with higher copies of amoA (a biomarker for enumerating nitrifiers), nirS and nirK (for denitrifiers), and nrfA (for DNRA bacteria). Among N-cycle processes, denitrification showed the highest activities and the highest concentrations of the corresponding gene (nirK and nirS) copy numbers. In riparian sediments, substantial nitrification activities (6.3 mg-N kg soil -1 d -1 average) and numbers of amoA copies (7.3 × 10 7  copies g soil -1 average) were observed, and nitrification rates correlate with denitrification rates. The guild structures of denitrifiers and nitrifiers in riparian sediments differed significantly from those found in stream sediments, as revealed by analysis of nirS and archaeal amoA sequences. This study shows that riparian sediments serve as sinks for inorganic nitrogen loads from non-point sources of agricultural runoff, with nitrification and denitrification associated with elevated levels of carbon and nitrogen contents and extracellular enzyme activities. Copyright © 2016 Elsevier Ltd. All rights reserved.

Denitrification and Biodiversity of Denitrifiers in a High-Mountain Mediterranean Lake

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Antonio Castellano-Hinojosa

2017-10-01

Full Text Available Wet deposition of reactive nitrogen (Nr species is considered a main factor contributing to N inputs, of which nitrate (NO3− is usually the major component in high-mountain lakes. The microbial group of denitrifiers are largely responsible for reduction of nitrate to molecular dinitrogen (N2 in terrestrial and aquatic ecosystems, but the role of denitrification in removal of contaminant nitrates in high-mountain lakes is not well understood. We have used the oligotrophic, high-altitude La Caldera lake in the Sierra Nevada range (Spain as a model to study the role of denitrification in nitrate removal. Dissolved inorganic Nr concentration in the water column of la Caldera, mainly nitrate, decreased over the ice-free season which was not associated with growth of microbial plankton or variations in the ultraviolet radiation. Denitrification activity, estimated as nitrous oxide (N2O production, was measured in the water column and in sediments of the lake, and had maximal values in the month of August. Relative abundance of denitrifying bacteria in sediments was studied by quantitative polymerase chain reaction of the 16S rRNA and the two phylogenetically distinct clades nosZI and nosZII genes encoding nitrous oxide reductases. Diversity of denitrifiers in sediments was assessed using a culture-dependent approach and after the construction of clone libraries employing the nosZI gene as a molecular marker. In addition to genera Polymorphum, Paracoccus, Azospirillum, Pseudomonas, Hyphomicrobium, Thauera, and Methylophaga, which were present in the clone libraries, Arthrobacter, Burkholderia, and Rhizobium were also detected in culture media that were not found in the clone libraries. Analysis of biological activities involved in the C, N, P, and S cycles from sediments revealed that nitrate was not a limiting nutrient in the lake, allowed N2O production and determined denitrifiers’ community structure. All these results indicate that

Denitrification and Biodiversity of Denitrifiers in a High-Mountain Mediterranean Lake

Science.gov (United States)

Castellano-Hinojosa, Antonio; Correa-Galeote, David; Carrillo, Presentación; Bedmar, Eulogio J.; Medina-Sánchez, Juan M.

2017-01-01

Wet deposition of reactive nitrogen (Nr) species is considered a main factor contributing to N inputs, of which nitrate (NO3−) is usually the major component in high-mountain lakes. The microbial group of denitrifiers are largely responsible for reduction of nitrate to molecular dinitrogen (N2) in terrestrial and aquatic ecosystems, but the role of denitrification in removal of contaminant nitrates in high-mountain lakes is not well understood. We have used the oligotrophic, high-altitude La Caldera lake in the Sierra Nevada range (Spain) as a model to study the role of denitrification in nitrate removal. Dissolved inorganic Nr concentration in the water column of la Caldera, mainly nitrate, decreased over the ice-free season which was not associated with growth of microbial plankton or variations in the ultraviolet radiation. Denitrification activity, estimated as nitrous oxide (N2O) production, was measured in the water column and in sediments of the lake, and had maximal values in the month of August. Relative abundance of denitrifying bacteria in sediments was studied by quantitative polymerase chain reaction of the 16S rRNA and the two phylogenetically distinct clades nosZI and nosZII genes encoding nitrous oxide reductases. Diversity of denitrifiers in sediments was assessed using a culture-dependent approach and after the construction of clone libraries employing the nosZI gene as a molecular marker. In addition to genera Polymorphum, Paracoccus, Azospirillum, Pseudomonas, Hyphomicrobium, Thauera, and Methylophaga, which were present in the clone libraries, Arthrobacter, Burkholderia, and Rhizobium were also detected in culture media that were not found in the clone libraries. Analysis of biological activities involved in the C, N, P, and S cycles from sediments revealed that nitrate was not a limiting nutrient in the lake, allowed N2O production and determined denitrifiers’ community structure. All these results indicate that denitrification could be a

Anaerobic columnar denitrification of high nitrate wastewater

International Nuclear Information System (INIS)

Francis, C.W.; Malone, C.D.

1975-01-01

Anaerobic columns were used to test the effectiveness of biological denitrification of nitrate solutions ranging in concentration from 1 to 10 kg NO 3 /m 3 . Several sources of nitrate (Ca(CNO 3 ) 2 , NaNO 3 , NH 4 NO 3 , and actual nitrate wastes from a UO 2 fuel fabrication plant) were evaluated as well as two packing media. The packing media were anthracite coal particles, whose effective diameter size ranged between 2 and 3 mm, and polypropylene Raschig rings 1.6 x 1.6 diameter. The anthracite coal proved to be the better packing media as excessive hydraulic short circuiting occurred in a 120 x 15 cm diameter glass column packed with the polypropylene rings after 40 days operation. With anthracite coal, floatation of the bed occurred at flow rates greater than 0.80 cm 3 /s. Tapered columns packed with anthracite coal eliminated the floatation problem, even at flow rates as high as 5 cm 3 /s. Under optimum operating conditions the anthracite coal behaved as a fluidized bed. Maximum denitrification rates were 1.0--1.4 g NO 3 /m 3 /s based on initial bed volume. Denitrification kinetics indicated that rates of denitrification became substrate inhibited at nitrate concentrations greater than 6.5 kg NO 3 /m 3 Anaerobic columns packed with anthracite coal appear to be an effective method of nitrate disposal for nitrate rich wastewater generated at UO 2 fuel fabrication plants and fuel reprocessing facilities. (U.S.)

Inactivation of ANAMMOX communities under concurrent operation of anaerobic ammonium oxidation (ANAMMOX) and denitrification

DEFF Research Database (Denmark)

Chamchoi, N.; Nitisoravut, S.; Schmidt, Jens Ejbye

2008-01-01

of Amx 820 and EUB 338 mixed. Denitrification was observed through the reductions of both COD and nitrate–nitrite concentrations under anaerobic/anoxic conditions. By providing a stoichiometric ratio of nitrite to ammonium nitrogen with addition nitrate nitrogen, a gradual reduction of ANAMMOX activity...... was found with an increase of COD concentration in a range of 100–400 mg l−1. This is equivalent to the COD to N ratio of 0.9–2.0. The COD concentration was found to be a control variable for process selection between ANAMMOX reaction and denitrification. A reduction of COD and nitrite–nitrate...

Modeling nitrous oxide production during biological nitrogen removal via nitrification and denitrification: extensions to the general ASM models.

Science.gov (United States)

Ni, Bing-Jie; Ruscalleda, Maël; Pellicer-Nàcher, Carles; Smets, Barth F

2011-09-15

Nitrous oxide (N(2)O) can be formed during biological nitrogen (N) removal processes. In this work, a mathematical model is developed that describes N(2)O production and consumption during activated sludge nitrification and denitrification. The well-known ASM process models are extended to capture N(2)O dynamics during both nitrification and denitrification in biological N removal. Six additional processes and three additional reactants, all involved in known biochemical reactions, have been added. The validity and applicability of the model is demonstrated by comparing simulations with experimental data on N(2)O production from four different mixed culture nitrification and denitrification reactor study reports. Modeling results confirm that hydroxylamine oxidation by ammonium oxidizers (AOB) occurs 10 times slower when NO(2)(-) participates as final electron acceptor compared to the oxic pathway. Among the four denitrification steps, the last one (N(2)O reduction to N(2)) seems to be inhibited first when O(2) is present. Overall, N(2)O production can account for 0.1-25% of the consumed N in different nitrification and denitrification systems, which can be well simulated by the proposed model. In conclusion, we provide a modeling structure, which adequately captures N(2)O dynamics in autotrophic nitrification and heterotrophic denitrification driven biological N removal processes and which can form the basis for ongoing refinements.

Photoperiodism and Enzyme Activity

Science.gov (United States)

Queiroz, Orlando; Morel, Claudine

1974-01-01

Metabolic readjustments after a change from long days to short days appear, in Kalanchoe blossfeldiana, to be achieved through the operation of two main mechanisms: variation in enzyme capacity, and circadian rhythmicity. After a lag time, capacity in phosphoenolpyruvate carboxylase and capacity in aspartate aminotransferase increase exponentially and appear to be allometrically linked during 50 to 60 short days; then a sudden fall takes place in the activity of the former. Malic enzyme and alanine aminotransferase behave differently. Thus, the operation of the two sections of the pathway (before and after the malate step) give rise to a continuously changing functional compartmentation in the pathway. Circadian rhythmicity, on the other hand, produces time compartmentation through phase shifts and variation in amplitude, independently for each enzyme. These characteristics suggest that the operation of a so-called biological clock would be involved. We propose the hypothesis that feedback regulation would be more accurate and efficient when applied to an already oscillating, clock-controlled enzyme system. PMID:16658749

Denitrification in the Arabian Sea

Digital Repository Service at National Institute of Oceanography (India)

Naqvi, S.W.A.; Noronha, R.J.; Reddy, C.V.G.

deficits because the approach requires denitrification to be restricted to Persian Gulf water. A method involving oxidative ratios has been modified and used to calculate nitrate deficits. The deficits so calculated lie between the values reported earlier...

Testing our understanding of Arctic denitrification using MIPAS-E satellite measurements in winter 2002/2003

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S. Davies

2006-01-01

Full Text Available Observations of gas-phase HNO3 and N2O in the polar stratosphere from the Michelson Interferometer for Passive Atmospheric Sounding aboard the ENVISAT satellite (MIPAS-E were made during the cold Arctic winter of 2002/2003. Vortex temperatures were unusually low in early winter and remained favourable for polar stratospheric cloud formation and denitrification until mid-January. MIPAS-E observations provide the first dataset with sufficient coverage of the polar vortex in mid-winter which enables a reasonable estimate of the timing of onset and spatial distribution of denitrification of the Arctic lower stratosphere to be performed. We use the observations from MIPAS-E to test the evolution of denitrification in the DLAPSE (Denitrification by Lagrangian Particle Sedimentation microphysical denitrification model coupled to the SLIMCAT chemical transport model. In addition, the predicted denitrification from a simple equilibrium nitric acid trihydrate-based scheme is also compared with MIPAS-E. Modelled denitrification is compared with in-vortex NOy and N2O observations from the balloon-borne MarkIV interferometer in mid-December. Denitrification was clearly observed by MIPAS-E in mid-December 2002 and reached 80% in the core of the vortex by early January 2003. The DLAPSE model is broadly able to capture both the timing of onset and the spatial distribution of the observed denitrification. A simple thermodynamic equilibrium scheme is able to reproduce the observed denitrification in the core of the vortex but overestimates denitrification closer to the vortex edge. This study also suggests that the onset of denitrification in simple thermodynamic schemes may be earlier than in the MIPAS-E observations.

Denitrification gene density across a wastewater-impacted riparian buffer zone

Science.gov (United States)

Riparian buffers are a best management practice used extensively to protect water bodies from agriculturally-generated nitrate pollution. In particular, the biological process of denitrification has been shown to be a sink for this nitrate. Denitrification results in the reduction of nitrate under a...

Nitrate removal in stream ecosystems measured by 15N addition experiments: Denitrification

Science.gov (United States)

Mulholland, P.J.; Hall, R.O.; Sobota, D.J.; Dodds, W.K.; Findlay, S.E.G.; Grimm, N. B.; Hamilton, S.K.; McDowell, W.H.; O'Brien, J. M.; Tank, J.L.; Ashkenas, L.R.; Cooper, L.W.; Dahm, Clifford N.; Gregory, S.V.; Johnson, S.L.; Meyer, J.L.; Peterson, B.J.; Poole, G.C.; Valett, H.M.; Webster, J.R.; Arango, C.P.; Beaulieu, J.J.; Bernot, M.J.; Burgin, A.J.; Crenshaw, C.L.; Helton, A.M.; Johnson, L.T.; Niederlehner, B.R.; Potter, J.D.; Sheibley, R.W.; Thomasn, S.M.

2009-01-01

We measured denitrification rates using a field 15N-NO- 3 tracer-addition approach in a large, cross-site study of nitrate uptake in reference, agricultural, and suburban-urban streams. We measured denitrification rates in 49 of 72 streams studied. Uptake length due to denitrification (SWden) ranged from 89 m to 184 km (median of 9050 m) and there were no significant differences among regions or land-use categories, likely because of the wide range of conditions within each region and land use. N2 production rates far exceeded N2O production rates in all streams. The fraction of total NO-3 removal from water due to denitrification ranged from 0.5% to 100% among streams (median of 16%), and was related to NHz 4 concentration and ecosystem respiration rate (ER). Multivariate approaches showed that the most important factors controlling SWden were specific discharge (discharge / width) and NO-3 concentration (positive effects), and ER and transient storage zones (negative effects). The relationship between areal denitrification rate (Uden) and NO- 3 concentration indicated a partial saturation effect. A power function with an exponent of 0.5 described this relationship better than a Michaelis-Menten equation. Although Uden increased with increasing NO- 3 concentration, the efficiency of NO-3 removal from water via denitrification declined, resulting in a smaller proportion of streamwater NO-3 load removed over a given length of stream. Regional differences in stream denitrification rates were small relative to the proximate factors of NO-3 concentration and ecosystem respiration rate, and land use was an important but indirect control on denitrification in streams, primarily via its effect on NO-3 concentration. ?? 2009.

Effects of de-icing salt on soil enzyme activity

Energy Technology Data Exchange (ETDEWEB)

Guentner, M; Wilke, B M

1983-01-01

Effects of de-icing salt on dehydrogenase, urease, alkalinephosphatase and arylsulfatase activity of O/sub L/- and A/sub h/-horizons of a moder and a mull soil were investigated using a field experiment. Additions of 2.5 kg m/sup -2/ and 5.0 kg m/sup -2/ of de-icing salt reduced activities of most enzymes within four weeks. Eleven months after salt addition there was nearly no reduction of enzyme activity to be measured on salt treated soils. The percentage of reduced enzyme activity was generally higher in the moder soil. It was concluded that reductions of enzyme activity were due to decreases of microbial activity and not to inactivation of enzymes.

Physiological levels of nitrate support anoxic growth by denitrification of Pseudomonas aeruginosa at growth rates reported in cystic fibrosis lungs and sputum

DEFF Research Database (Denmark)

Line, Laura; Alhede, Morten; Kolpen, Mette

2014-01-01

denitrification. The growth rate of P. aeruginosa achieved by denitrification at physiological levels (~400 μM) of nitrate (NO(-) 3) is however, not known. Therefore, we have measured growth rates of anoxic cultures of PAO1 and clinical isolates (n = 12) in LB media supplemented with NO(-) 3 and found...... a significant increase of growth when supplementing PAO1 and clinical isolates with ≥150 μM NO(-) 3 and 100 μM NO(-) 3, respectively. An essential contribution to growth by denitrification was demonstrated by the inability to establish a significantly increased growth rate by a denitrification deficient Δnir...... of the four N-oxide reductases in PAO1 (Nar, Nir, Nor, Nos) further verified the engagement of denitrification, showing a transient increase in activation and expression and rapid consumption of NO(-) 3 followed by a transient increase of NO(-) 2. Growth rates obtained by denitrification in this study were...

[Interaction between CYP450 enzymes and metabolism of traditional Chinese medicine as well as enzyme activity assay].

Science.gov (United States)

Lu, Tu-lin; Su, Lian-lin; Ji, De; Gu, Wei; Mao, Chun-qin

2015-09-01

Drugs are exogenous compounds for human bodies, and will be metabolized by many enzymes after administration. CYP450 enzyme, as a major metabolic enzyme, is an important phase I drug metabolizing enzyme. In human bodies, about 75% of drug metabolism is conducted by CYP450 enzymes, and CYP450 enzymes is the key factor for drug interactions between traditional Chinese medicine( TCM) -TCM, TCM-medicine and other drug combination. In order to make clear the interaction between metabolic enzymes and TCM metabolism, we generally chose the enzymatic activity as an evaluation index. That is to say, the enhancement or reduction of CYP450 enzyme activity was used to infer the inducing or inhibitory effect of active ingredients and extracts of traditional Chinese medicine on enzymes. At present, the common method for measuring metabolic enzyme activity is Cocktail probe drugs, and it is the key to select the suitable probe substrates. This is of great significance for study drug's absorption, distribution, metabolism and excretion (ADME) process in organisms. The study focuses on the interaction between TCMs, active ingredients, herbal extracts, cocktail probe substrates as well as CYP450 enzymes, in order to guide future studies.

Compounds from silicones alter enzyme activity in curing barnacle glue and model enzymes.

Science.gov (United States)

Rittschof, Daniel; Orihuela, Beatriz; Harder, Tilmann; Stafslien, Shane; Chisholm, Bret; Dickinson, Gary H

2011-02-17

Attachment strength of fouling organisms on silicone coatings is low. We hypothesized that low attachment strength on silicones is, in part, due to the interaction of surface available components with natural glues. Components could alter curing of glues through bulk changes or specifically through altered enzyme activity. GC-MS analysis of silicone coatings showed surface-available siloxanes when the coatings were gently rubbed with a cotton swab for 15 seconds or given a 30 second rinse with methanol. Mixtures of compounds were found on 2 commercial and 8 model silicone coatings. The hypothesis that silicone components alter glue curing enzymes was tested with curing barnacle glue and with commercial enzymes. In our model, barnacle glue curing involves trypsin-like serine protease(s), which activate enzymes and structural proteins, and a transglutaminase which cross-links glue proteins. Transglutaminase activity was significantly altered upon exposure of curing glue from individual barnacles to silicone eluates. Activity of purified trypsin and, to a greater extent, transglutaminase was significantly altered by relevant concentrations of silicone polymer constituents. Surface-associated silicone compounds can disrupt glue curing and alter enzyme properties. Altered curing of natural glues has potential in fouling management.

Measurement of enzyme activity.

Science.gov (United States)

Harris, T K; Keshwani, M M

2009-01-01

To study and understand the nature of living cells, scientists have continually employed traditional biochemical techniques aimed to fractionate and characterize a designated network of macromolecular components required to carry out a particular cellular function. At the most rudimentary level, cellular functions ultimately entail rapid chemical transformations that otherwise would not occur in the physiological environment of the cell. The term enzyme is used to singularly designate a macromolecular gene product that specifically and greatly enhances the rate of a chemical transformation. Purification and characterization of individual and collective groups of enzymes has been and will remain essential toward advancement of the molecular biological sciences; and developing and utilizing enzyme reaction assays is central to this mission. First, basic kinetic principles are described for understanding chemical reaction rates and the catalytic effects of enzymes on such rates. Then, a number of methods are described for measuring enzyme-catalyzed reaction rates, which mainly differ with regard to techniques used to detect and quantify concentration changes of given reactants or products. Finally, short commentary is given toward formulation of reaction mixtures used to measure enzyme activity. Whereas a comprehensive treatment of enzymatic reaction assays is not within the scope of this chapter, the very core principles that are presented should enable new researchers to better understand the logic and utility of any given enzymatic assay that becomes of interest.

Denitrification nitrogen gas formation and gene expression in alpine grassland soil as affected by climate change conditions

Science.gov (United States)

Chen, Zhe; Wang, Changhui; Gschwendtner, Silvia; Schloter, Michael; Butterbach-Bahl, Klaus; Dannenmann, Michael

2013-04-01

that denitrification N gas formation accounted for 21%, dissimilatory nitrate reduction to ammonium for 8%, and microbial immobilization for 73% of nitrate consumption. Overall, our study shows that changes in climate exert feedback on denitrification N gas formation and N gas product ratios via changes in microbial activity at the level of single denitrification steps. The close relationships found between denitrification N gas formation, N gas product ratios and denitrification gene expression suggests a large potential of molecular methods to predict denitrification dynamics in soil.

Nutrient and carbon availability influences on denitrification in the regulated Lower Colorado River, Austin

Science.gov (United States)

Spector, J.

2016-12-01

The Lower Colorado River in Austin, Texas receives nitrogen-rich runoff and treated wastewater effluent and is subject to periodic water releases from the Longhorn Dam, which cause fluctuations in groundwater stage downstream. This research examined groundwater denitrification at the Hornsby Bend riparian area (located approximately 24 km downstream of downtown Austin) and characterized how dam-induced hyporheic exchange affects denitrification rates. Conductivity, temperature, water level, and dissolved oxygen concentrations were measured continuously throughout flood pulses for six months using dataloggers installed in a transect of seven monitoring wells on the river bank. Hourly samples were collected using an autosampler in one monitoring well (MW-5) during various flood conditions during the six month monitoring period. Water samples were analyzed for total organic carbon, total nitrogen, anions (NO3- and NO2-), NH4+ concentrations, alkalinity, and specific ultraviolet absorbance (SUVA) to characterize dissolved organic matter. Following large flood events (up to 4 m of water level stage increase), average conductivity increased 300 µs/centimeter in MW-5 as the water level receded. Analysis of water samples indicated that NO3- reduction occurred as conductivity and alkalinity increased. In addition, NH4+ concentrations increased during high conductivity periods. Increased denitrification activity corresponded with high SUVA. High conductivity and alkalinity increase the availability of electron donors (HCO3- and CO32-) and enhances denitrification potential. Higher SUVA values indicate increased dissolved organic carbon aromaticity and corresponding NO3- reduction. Additionally, changes in dissolved organic matter lability indicate the residence times of possible reactive organic carbon in the riparian area. This study has implications for determining advantageous geochemical conditions for hyporheic zone denitrification following large flood events.

Use of microbial analysis to evaluate denitrification in the karstic aquifer of Okinawa, Japan

Science.gov (United States)

Yasumoto, J.

2014-12-01

Denitrification, a microbial process in the nitrogen cycle, is a facultative respiratory pathway in which nitrate (NO3-), nitrite (NO2-), nitric oxide (NO), and nitrous oxide (N2O), successively, are reduced to nitrogen gas (N2). This study explores the use of microbial analysis to evaluate the processes involved in nitrate attenuation in groundwater. Polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) is used to identify denitrifiers based only on their 16SrRNA gene sequences, and Real-Time PCR analysis is used to quantify nitrite reducing genes (nirK and nirS), this suggest that a new methods for detecting denitrification activity by comparing the gene dosage that has been detected by RT-PCR and the value of the δ15NNO3- and δ18ONO3-. This study focuses on a zone of significant NO3- attenuation occurring at underground dam catchment area in the karstic Ryukyu limestone aquifer, which is located southern part of Okinawa, Japan. As a result of microbial analysis, the bacteria were detected at all observation points which have been reported to have denitrification ability. And it has been confirmed that the bacteria has a gene nirS which is related to denitrification. In addition, many bacteria related to denitrification have been extracted from suspended solids more than from groundwater in the aquifer. And, the correlation was high between nirK /nirS gene dosage that has been detected by RT-PCR and the value of the δ15N and δ18O; therefore, this study demonstrates the effectiveness of using Real-Time PCR analysis for providing insights into the processes affecting nitrate attenuation in ground water.

Denitrification as the dominant nitrogen loss process in the Arabian Sea

Digital Repository Service at National Institute of Oceanography (India)

Ward, B.B.; Devol, A.H.; Rich, J.J.; Chang, B.X.; Bulow, S.E.; Naik, H.; Pratihary, A.K.; Jayakumar, A.

experiments in the Arabian Sea denitrification is responsible for 87-99% of the total N sub(2) production. The dominance of denitrification is reproducible using two independent isotope incubation methods. In contrast, anammox is dominant in the Eastern...

Enzyme activity and kinetics in substrate-amended river sediments

Energy Technology Data Exchange (ETDEWEB)

Duddridge, J E; Wainwright, M

1982-01-01

In determining the effects of heavy metals in microbial activity and litter degradation in river sediments, one approach is to determine the effects of these pollutants on sediment enzyme activity and synthesis. Methods to assay amylase, cellulase and urease activity in diverse river sediments are reported. Enzyme activity was low in non-amended sediments, but increased markedly when the appropriate substrate was added, paralleling both athropogenic and natural amendment. Linear relationships between enzyme activity, length of incubation, sample size and substrate concentration were established. Sediment enzyme activity generally obeyed Michaelis-Menton kinetics, but of the three enzymes, urease gave least significant correlation coefficients when the data for substrate concentration versus activity was applied to the Eadie-Hofstee transformation of the Michaelis-Menten equation. K/sub m/ and V/sub max/ for amylase, cellulase and urease in sediments are reported. (JMT)

Enzyme Activity Experiments Using a Simple Spectrophotometer

Science.gov (United States)

Hurlbut, Jeffrey A.; And Others

1977-01-01

Experimental procedures for studying enzyme activity using a Spectronic 20 spectrophotometer are described. The experiments demonstrate the effect of pH, temperature, and inhibitors on enzyme activity and allow the determination of Km, Vmax, and Kcat. These procedures are designed for teaching large lower-level biochemistry classes. (MR)

The development and application of SCR denitrification technology in power plant

Science.gov (United States)

Wu, Junnan

2017-12-01

In recent decades, the emission of the nitrogen oxides (NOX) has been increasing with the years of the thermal power plant. The environment pollution caused by the emission of quantities of nitrogen oxides became more and more serious, so people now put more emphasis on the control of the emission of the nitrogen oxides. Especially, our country and the society are paying much more attention to the environment protection and the environment problems cannot be neglected. In this paper, we introduced the related research background of the technology of SCR denitrification which was as the symbol of the technology of the catalytic denitrification and discussed the reaction principles of the SCR denitrification and frequently used catalysts, the process of the technology, and the configuration. In the end, we pointed the way of the future research of the technology of the SCR denitrification.

Activation of interfacial enzymes at membrane surfaces

DEFF Research Database (Denmark)

Mouritsen, Ole G.; Andresen, Thomas Lars; Halperin, Avi

2006-01-01

A host of water-soluble enzymes are active at membrane surfaces and in association with membranes. Some of these enzymes are involved in signalling and in modification and remodelling of the membranes. A special class of enzymes, the phospholipases, and in particular secretory phospholipase A2 (s...

Screening identification of aerobic denitrification bacteria with high soil desalinization capacity

Science.gov (United States)

Jin, H.; Chen, H.; Jin, H.; Qian, Y.; Zhang, K.

2017-08-01

In order to study the mechanism of bacteria used in the saline soil remediation process, the aerobic denitrification bacteria were isolated from an agricultural greenhouse soil in a farm in East China’s Zhejiang Province. The identification, nitrogen reducing characteristics and the denitrification effect of bacteria from different soils at various locations were investigated. The results showed that the NO3- removal rate was 91% with bacteria from the greenhouse soil under aerobic conditions in 52 h, and the bacteria were identified as Gram-positive Castellaniella denitrification bacteria.

Activity assessment of microbial fibrinolytic enzymes.

Science.gov (United States)

Kotb, Essam

2013-08-01

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

Operation of a fluidized-bed bioreactor for denitrification

International Nuclear Information System (INIS)

Hancher, C.W.; Taylor, P.A.; Napier, J.M.

1978-01-01

Two denitrification fluidized-bed bioreactors of the same length (i.e., 5 m) but with different inside diameters (i.e., 5 and 10 cm) have been operated on feed ranging in nitrate concentration from 200 to 2000 g/m 3 ; thus far, good agreement has been obtained. Two 10-cm-ID bioreactors operating in series have also been tested; the results are in accordance with predicted results based on the performance of a 5-cm-ID bioreactor. The overall denitrification rate in the dual 10-cm-ID bioreactor system was found to be 23 kg N(NO 3 - )/day-m 3 using feed with a nitrate concentration of 1800 g/m 3 . Data obtained in operating-temperature tests indicate that the maximum denitrification rate is achieved between 22 and 30 0 C. These data will form the basis of the design of our mobile pilot plant which consists of dual 20-cm-ID by 7.3-m-long bioreactors

Denitrification: An important pathway for nitrous oxide production in tropical mangrove sediments (Goa, India)

Digital Repository Service at National Institute of Oceanography (India)

Fernandes, S.O.; LokaBharathi, P.A.; Bonin, P.C.; Michotey, V.D.

Net nitrous oxide production and denitrification activity were measured in two mangrove ecosystems of Goa, India. The relatively pristine site Tuvem was compared to Divar, which is prone to high nutrient input. Stratified sampling at 2-cm intervals...

Detection of enzyme activity in decontaminated spices of industrial use

International Nuclear Information System (INIS)

Müller, R.; Theobald, R.

1995-01-01

A range of decontaminated spices of industrial use have been examinated for their enzymes (catalase, peroxidase, amylase, lipase activity). The genuine enzymes remain fully active in irradiated spices, whereas the microbial load is clearly reduced. In contrast steam treated spices no longer demonstrate enzyme activities. Steam treatment offers e.g. black pepper without lipase activity, which can no longer cause fat deterioration. Low microbial load in combination with clearly detectable enzyme activity in spices is an indication for irradiation, whereas, reduced microbial contamination combined with enzyme inactivation indicate steam treatment of raw material [de

Design of fluidized-bed, biological denitrification systems

International Nuclear Information System (INIS)

Patton, B.D.; Hancher, C.W.; Pitt, W.W.; Walker, J.F.

1982-01-01

Many commercial processes yield nitrate-containing wastewaters that are being discharged to the environment because traditional recovery or disposal methods are economically unacceptable. The anticipated discharge limits (i.e., 10 to 20 g (NO 3 - )/m 3 ) being considered by many states will not allow continued release of these wastewaters. The new discharge standards can be met economically by use of the fluidizied-bed, biological denitrification process. Research and development studies were conducted with 0.05-, 0.10-, 0.20-, and 0.50-m-diam fluidized-bed bioreactor systems. Feed nitrate concentrations were in the 0 to 10,000 g (NO 3 - )/m 3 range. Using the data from these studies, rate expressions were developed for the destruction of nitrate as a function of nitrate concentration. Methods were also developed for sizing bioreactors and biomass control systems. The sizing methods for fluidized-bed denitrification systems are described, and support systems such as sampling and analysis, instrumentation and controls, utilities, and bacteria storage are discussed. Operation of the process is also briefly discussed to aid the designer. Using the methods presented in this report, fluidized-bed, biological denitrification systems can be designed to treat nitrate wastewater streams

Denitrification and potential nitrous oxide and carbon dioxide production in brownfield wetland soils.

Science.gov (United States)

Palta, Monica M; Ehrenfeld, Joan G; Groffman, Peter M

2013-09-01

Brownfields, previously developed sites that are derelict, vacant, or underused, are ubiquitous in urban areas. Wetlands on brownfields often retain rain and stormwater longer than the surrounding landscape because they are low-lying; this increases the possibility for these areas to process waterborne contaminants from the urban environment. In the northeastern United States, atmospheric deposition of nitrate (NO) is high. Denitrification, a microbial process common in wetlands, is a means of removing excess NO. Nitrogen gas is the desired end product of denitrification, but incomplete denitrification results in the production of NO, a greenhouse gas. The goal of this study was to investigate the potential of brownfield wetlands to serve as sinks for inorganic nitrogen and sources of greenhouse gases. We examined limitations to denitrification and NO production in brownfield wetland soils in New Jersey. Soil C:N ratios were high (18-40) and intact core denitrification (-0.78 to 11.6 μg NO-N kg dry soil d) and N mineralization (0.11-2.97 mg N kg dry soil d) were low for all sites. However, soil NO increased during dry periods. Nitrate additions to soil slurries increased denitrification rates, whereas labile C additions did not, indicating that soil denitrifiers were nitrogen limited. Incubations indicated that the end product of denitrification was primarily NO and not N. These results indicate that brownfield wetlands can develop significant denitrification capacity, potentially causing NO limitation. They might be significant sinks for atmospheric NO but may also become a significant source of NO if NO deposition were to increase. Copyright © by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America, Inc.

Sequential nitrification and denitrification in a novel palm shell granular activated carbon twin-chamber upflow bio-electrochemical reactor for treating ammonium-rich wastewater.

Science.gov (United States)

Mousavi, Seyyedalireza; Ibrahim, Shaliza; Aroua, Mohamed Kheireddine

2012-12-01

In this study, a twin-chamber upflow bio-electrochemical reactor packed with palm shell granular activated carbon as biocarrier and third electrode was used for sequential nitrification and denitrification of nitrogen-rich wastewater under different operating conditions. The experiments were performed at a constant pH value for the denitrification compartment. The effect of variables, namely, electric current (I) and hydraulic retention time (HRT), on the pH was considered in the nitrification chamber. The response surface methodology was used based on three levels to develop empirical models for the study on the effects of HRT and current values as independent operating variables on NH(4)(+)-N removal. The results showed that ammonium was reduced within the function of an extensive operational range of electric intensity (20-50 mA) and HRT (6-24h). The optimum condition for ammonium oxidation (90%) was determined with an I of 32 mA and HRT of 19.2h. Copyright © 2012 Elsevier Ltd. All rights reserved.

Effect of irradiation on lysosomal enzyme activation in cultured macrophages

International Nuclear Information System (INIS)

Clarke, C.; Wills, E.D.

1980-01-01

The effect of γrays on lysosomal enzyme activity of normal and immune macrophages of DBA/2 mice cultured in vitro has been studied. A dose of 500 rad did not significantly affect lysosomal enzyme activity 3 hours after irradiation but caused the activity to increase to 1.4 times the control value 22.5 hours after irradiation. 22.5 hours after a dose of 3000 rad the enzyme activity increased to 2.5 times the control. Lysosomal enzyme activity of the macrophages was also markedly increased by immunization of the mice with D lymphoma cells, before culture in vitro, but irradiation of these cells with a dose of 500 rad caused a further increase in lysosomal enzyme activity. The results indicate that immunization and irradiation both cause stimulation of lysosomal enzyme activity in macrophages but that the mechanisms of activation are unlikely to be identical. (author)

Evaluating vertical concentration profile of carbon source released from slow-releasing carbon source tablets and in situ biological nitrate denitrification activity

Science.gov (United States)

Yeum, Y.; HAN, K.; Yoon, J.; Lee, J. H.; Song, K.; Kang, J. H.; Park, C. W.; Kwon, S.; Kim, Y.

2017-12-01

Slow-releasing carbon source tablets were manufactured during the design of a small-scale in situ biological denitrification system to reduce high-strength nitrate (> 30 mg N/L) from a point source such as livestock complexes. Two types of slow-releasing tablets, precipitating tablet (PT, apparent density of 2.0 g/mL) and floating tablet (FT), were prepared to achieve a vertically even distribution of carbon source (CS) in a well and an aquifer. Hydroxypropyl methylcellulose (HPMC) was used to control the release rate, and microcrystalline cellulose pH 101 (MCC 101) was added as a binder. The #8 sand was used as a precipitation agent for the PTs, and the floating agents for the FTs were calcium carbonate and citric acid. FTs floated within 30 min. and remained in water because of the buoyance from carbon dioxide, which formed during the acid-base reaction between citric acid and calcium carbonate. The longevities of PTs with 300 mg of HPMC and FTs with 400 mg of HPMC were 25.4 days and 37.3 days, respectively. We assessed vertical CS profile in a continuous flowing physical aquifer model (release test, RT) and its efficiency on biological nitrate denitrification (denitrification test, DT). During the RT, PTs, FTs and a tracer (as 1 mg rhodamine B/L) were initially injected into a well of physical aquifer model (PAM). Concentrations of CS and the tracer were monitored along the streamline in the PAM to evaluate vertical profile of CS. During the DT, the same experiment was performed as RT, except continuous injection of solution containing 30 mg N/L into the PAM to evaluate biological denitrification activity. As a result of RT, temporal profiles of CS were similar at 3 different depths of monitoring wells. These results suggest that simultaneous addition of PT and FT be suitable for achieving a vertically even distribution of the CS in the injection well and an aquifer. In DT, similar profile of CS was detected in the injection well, and nitrate was biologically

Denitrification of fertilizer wastewater at high chloride concentration

DEFF Research Database (Denmark)

Ucisik, Ahmed Süheyl; Henze, Mogens

Wastewater from fertilizer industry is characterized by high contents of chloride concentration, which normally vary between 60 and 76 g/l. Experiments with bilogical denitrification were performed in lab-scale "fill and draw" reactors with synthetic wastewater with chloride concentrations up to 77.......4 g/l. The results of the experiments showed that biological denitrification was feasible at the extreme environmental conditions prevailing in fertilizer wastewater. Stable continuous biological denitrfication of the synthetic high chloride wastewater was performed up to 77.4 g Cl/l at 37 degree C...

Enzyme specific activity in functionalized nanoporous supports

International Nuclear Information System (INIS)

Lei Chenghong; Soares, Thereza A; Shin, Yongsoon; Liu Jun; Ackerman, Eric J

2008-01-01

Here we reveal that enzyme specific activity can be increased substantially by changing the protein loading density (P LD ) in functionalized nanoporous supports so that the enzyme immobilization efficiency (I e , defined as the ratio of the specific activity of the immobilized enzyme to the specific activity of the free enzyme in solution) can be much higher than 100%. A net negatively charged glucose oxidase (GOX) and a net positively charged organophosphorus hydrolase (OPH) were entrapped spontaneously in NH 2 - and HOOC-functionalized mesoporous silica (300 A, FMS) respectively. The specific activity of GOX entrapped in FMS increased with decreasing P LD . With decreasing P LD , I e of GOX in FMS increased from 150%. Unlike GOX, OPH in HOOC-FMS showed increased specific activity with increasing P LD . With increasing P LD , the corresponding I e of OPH in FMS increased from 100% to>200%. A protein structure-based analysis of the protein surface charges directing the electrostatic interaction-based orientation of the protein molecules in FMS demonstrates that substrate access to GOX molecules in FMS is limited at high P LD , consequently lowering the GOX specific activity. In contrast, substrate access to OPH molecules in FMS remains open at high P LD and may promote a more favorable confinement environment that enhances the OPH activity

Heterotrophic nitrification and aerobic denitrification bacterium ...

African Journals Online (AJOL)

Jane

2011-07-18

Jul 18, 2011 ... nutrient agar plates 3 times from the enrichment liquid medium. The isolates ..... growing in the logarithmic phase, the decomposition rate was higher than .... denitrification in bench-scale sequencing batch reactors. Water Res.

EVOLUTIONARY TRANSITIONS IN ENZYME ACTIVITY OF ANT FUNGUS GARDENS

DEFF Research Database (Denmark)

De Fine Licht, Henrik H; Schiøtt, Morten; Mueller, Ulrich G

2010-01-01

an association with a monophyletic clade of specialized symbionts. In conjunction with the transition to specialized symbionts, the ants advanced in colony size and social complexity. Here we provide a comparative study of the functional specialization in extracellular enzyme activities in fungus gardens across...... the attine phylogeny. We show that, relative to sister clades, gardens of higher-attine ants have enhanced activity of protein-digesting enzymes, whereas gardens of leaf-cutting ants also have increased activity of starch-digesting enzymes. However, the enzyme activities of lower-attine fungus gardens...... are targeted primarily towards partial degradation of plant cell walls, reflecting a plesiomorphic state of non-domesticated fungi. The enzyme profiles of the higher-attine and leaf-cutting gardens appear particularly suited to digest fresh plant materials and to access nutrients from live cells without major...

Simulation of three-phase fluidized bioreactors for denitrification

International Nuclear Information System (INIS)

Hamza, A.V.; Dolan, J.F.; Wong, E.W.

1981-03-01

Fluidized-bed bioreactors were developed and operated at three scales (diameters of 0.1, 0.2, and 0.5 m) by the Chemical Technology Division. The performance of these reactors in denitrification was simulated using the following modified form of Monod kinetics to describe the reaction kinetics: rate = V/sub max/ (NO 3 - /K/sub s/ + NO 3 - ) (% biomass). In the fluids-movement portion of the simulation the tanks-in-series approximation to backmixing was used. This approach yielded a V/sub max/ of 3.5 g/m 3 -min (% biomass) and a K/sub s/ of 163 g/m 3 for the 0.5-m bioreactor. Values of V/sub max/ and K/sub s/ were also determined for data derived from the 0.1-m bioreactor, but inadequate RTD data reduced the confidence level in these results. A complication in denitrification is the multi-step nature of the reduction from nitrate to nitrite to hyponitrite and finally to nitrogen. An experimental study of the effect of biomass loading upon denitrification was begun. It is recommended that the experimental work be continued

Denitrification in the Arabian Sea: A 3D ecosystem modelling study

Science.gov (United States)

Anderson, Thomas R.; Ryabchenko, Vladimir A.; Fasham, Michael J. R.; Gorchakov, Victor A.

2007-12-01

A three-dimensional hydrodynamic-ecosystem model was used to examine the factors determining the spatio-temporal distribution of denitrification in the Arabian Sea. The ecosystem model includes carbon and nitrogen as currencies, cycling of organic matter via detritus and dissolved organic matter, and both remineralization and denitrification as sinks for material exported below the euphotic zone. Model results captured the marked seasonality in plankton dynamics of the region, with characteristic blooms of chlorophyll in the coastal upwelling regions and central Arabian Sea during the southwest monsoon, and also in the northern Arabian Sea during the northeast monsoon as the mixed layer shoals. Predicted denitrification was 26.2 Tg N yr -1,the greatest seasonal contribution being during the northeast monsoon when primary production is co-located with the zone of anoxia. Detritus was the primary organic substrate consumed in denitrification (97%), with a small (3%) contribution by dissolved organic matter. Denitrification in the oxygen minimum zone was predicted to be fuelled almost entirely by organic matter supplied by particles sinking vertically from the euphotic zone above (0.73 mmol N m -2 d -1) rather than from lateral transport of organic matter from elsewhere in the Arabian Sea (less than 0.01 mmol N m -2 d -1). Analysis of the carbon budget in the zone of denitrification (north of 10°N and east of 55°E) indicates that the modelled vertical export flux of detritus, which is similar in magnitude to estimates from field data based on the 234Th method, is sufficient to account for measured bacterial production below the euphotic zone in the Arabian Sea.

Denitrification coupled with methane anoxic oxidation and microbial community involved identification

Directory of Open Access Journals (Sweden)

Renata Medici Frayne Cuba

2011-02-01

Full Text Available In this work, the biological denitrification associated with anoxic oxidation of methane and the microbial diversity involved were studied. Kinetic tests for nitrate (NO3- and nitrite (NO2- removal and methane uptake were carried out in 100 mL batch reactors incubated in a shaker (40 rpm at 30 ºC. Denitrificant/methanotrophic biomass was taken from a laboratory scale reactor fed with synthetic nitrified substrates (40 mgN L-1 of NO3- and subsequently NO2- and methane as carbon source. Results obtained from nitrate removal followed a first order reaction, presenting a kinetic apparent constant (kNO3 of 0.0577±0.0057d-1. Two notable points of the denitrification rate (0.12gNO3--N g-1 AVS d-1 and 0.07gNO3--N g-1 AVS d-1 were observed in the beginning and on the seventh day of operation. When nitrite was added as an electron acceptor, denitrification rates were improved, presenting an apparent kinetic constant (kNO2 of 0.0722±0.0044d-1, a maximum denitrification rate of 0.6gNO2--N g-1AVS d-1, and minimum denitrification rate of 0.1gNO2--N g-1AVS d-1 at the beginning and end of the test, respectively. Endogenous material supporting denitrification and methane concentration dissolved in the substrate was discarded from the control experiments in the absence of methane and seed, respectively. Methylomonas sp. was identified in the reactors fed with nitrate and nitrite as well as uncultured bacterium.

Detection of Extracellular Enzyme Activities in Ganoderma neo-japonicum

OpenAIRE

Jo, Woo-Sik; Park, Ha-Na; Cho, Doo-Hyun; Yoo, Young-Bok; Park, Seung-Chun

2011-01-01

The ability of Ganoderma to produce extracellular enzymes, including β-glucosidase, cellulase, avicelase, pectinase, xylanase, protease, amylase, and ligninase was tested in chromogenic media. β-glucosidase showed the highest activity, among the eight tested enzymes. In particular, Ganoderma neo-japonicum showed significantly stronger activity for β-glucosidase than that of the other enzymes. Two Ganoderma lucidum isolates showed moderate activity for avicelase; however, Ganoderma neo-japonic...

Denitrification and Ecosystem Services: Mapping and Modeling Conservation Effects

Science.gov (United States)

Morris, C. K.; Walter, T.

2012-12-01

Precision conservation is the latest effort to increase higher efficiency in agricultural best management practices by considering the spatial and temporal variability in agroecosystems. The authors have developed a framework for incorporating the ecosystem service of denitrification into an existing precision conservation mapping tool. The model identifies areas of denitirification and quantifies potential denitrification when a conservation practice is adopted. The methodology is being tested in a small subwatershed in the Upper Susquehanna Basin of New York State.

An appraisal of the enzyme stability-activity trade-off.

Science.gov (United States)

Miller, Scott R

2017-07-01

A longstanding idea in evolutionary physiology is that an enzyme cannot jointly optimize performance at both high and low temperatures due to a trade-off between stability and activity. Although a stability-activity trade-off has been observed for well-characterized examples, such a trade-off is not imposed by any physical chemical constraint. To better understand the pervasiveness of this trade-off, I investigated the stability-activity relationship for comparative biochemical studies of purified orthologous enzymes identified by a literature search. The nature of this relationship varied greatly among studies. Notably, studies of enzymes with low mean synonymous nucleotide sequence divergence were less likely to exhibit the predicted negative correlation between stability and activity. Similarly, a survey of directed evolution investigations of the stability-activity relationship indicated that these traits are often uncoupled among nearly identical yet phenotypically divergent enzymes. This suggests that the presumptive trade-off often reported for investigations of enzymes with high mean sequence divergence may in some cases instead be a consequence of the degeneration over time of enzyme function in unselected environments, rather than a direct effect of thermal adaptation. The results caution against the general assertion of a stability-activity trade-off during enzyme adaptation. © 2017 The Author(s). Evolution © 2017 The Society for the Study of Evolution.

Comparison of Simultaneous Nitrification and Denitrification for Three Different Reactors

Directory of Open Access Journals (Sweden)

W. Khanitchaidecha

2015-01-01

Full Text Available Discharge of high NH4-N containing wastewater into water bodies has become a critical and serious issue due to its negative impact on water and environmental quality. In this research, the performance of three different reactors was assessed and compared with regard to the removal of NH4-N from wastewater. The highest nitrogen removal efficiency of 98.3% was found when the entrapped sludge reactor (ESR, in which the sludge was entrapped in polyethylene glycol polymer, was used. Under intermittent aeration, nitrification and denitrification occurred simultaneously in the aerobic and anaerobic periods. Moreover, internal carbon was consumed efficiently for denitrification. On the other hand, internal carbon consumption was not found to occur in the suspended sludge reactor (SSR and the mixed sludge reactor (MSR and this resulted in nitrogen removal efficiencies of SSR and MSR being 64.7 and 45.1%, respectively. Nitrification and denitrification were the main nitrogen removal processes in the aerobic and anaerobic periods, respectively. However, due to the absence of sufficient organic carbon, denitrification was uncompleted resulting in high NO3-N contents in the effluent.

Denitrification capacity and greenhouse gas emissions of soils in channelized and restored reaches along an Alpine river corridor

Science.gov (United States)

Shrestha, Juna; Niklaus, Pascal; Samaritani, Emanuela; Frossard, Emmanuel; Tockner, Klement; Luster, Jörg

2010-05-01

In order to assess the effects of river restoration on water and air quality, the biogeochemical functions of channelized and restored river reaches have to be quantified. The objective of this study was to compare denitrification potential and greenhouse gas emissions of functional processing zones (FPZ) in a channelized and a recently restored reach of the alpine river Thur in north-eastern Switzerland. The study was part of the project cluster RECORD of the ETH domain, Switzerland, which was initiated to increase the mechanistic understanding of coupled hydrological and ecological processes in river corridors. The denitrification potential represents an important aspect of the soil filter function related to water quality. Besides, it also contributes to the emission of greenhouse gases. Extensively used pasture growing on a sandy loam is the characteristic FPZ of the channelized section. The restored section encompasses five FPZ: (i) bare gravel bars sparsely colonized by plants, (ii) gravel bars densely colonized by grass (mainly canary reed grass with up to 80 cm sandy deposits), (iii) mixed forest dominated by ash and maple, (iv) riparian forest dominated by willow (Salix alba), (v) older overbank sediments stabilized during restoration with young willows separating the forests from the river-gravel bar system (willow bush). The FPZ were sampled in January, April, August and October 2009. In addition, in June and July 2009 two flood events were monitored in the restored section with more frequent samplings. At each date, topsoil samples were collected in each FPZ (four replicates per samples) and analyzed for denitrifier enzyme activity (DEA). In addition, gas samples were taken in-situ using the closed chamber technique to measure soil respiration as well as N2O and CH4 fluxes. In all FPZ, the denitrification potential was mainly governed by soil moisture. It was highest in the willow forest exhibiting low spatial variability. The DEA in pasture, grass zone

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

DEFF Research Database (Denmark)

Hendriksen, Niels Bohse; Winding, Anne

2012-01-01

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

Marine denitrification rates determined from a global 3-D inverse model

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T. DeVries

2013-04-01

Full Text Available A major impediment to understanding long-term changes in the marine nitrogen (N cycle is the persistent uncertainty about the rates, distribution, and sensitivity of its largest fluxes in the modern ocean. We use a global ocean circulation model to obtain the first 3-D estimate of marine denitrification rates that is maximally consistent with available observations of nitrate deficits and the nitrogen isotopic ratio of oceanic nitrate. We find a global rate of marine denitrification in suboxic waters and sediments of 120–240 Tg N yr−1, which is lower than many other recent estimates. The difference stems from the ability to represent the 3-D spatial structure of suboxic zones, where denitrification rates of 50–77 Tg N yr−1 result in up to 50% depletion of nitrate. This depletion reduces the effect of local isotopic enrichment on the rest of the ocean, allowing the N isotope ratio of oceanic nitrate to be achieved with a sedimentary denitrification rate about 1.3–2.3 times that of suboxic zones. This balance of N losses between sediments and suboxic zones is shown to obey a simple relationship between isotope fractionation and the degree of nitrate consumption in the core of the suboxic zones. The global denitrification rates derived here suggest that the marine nitrogen budget is likely close to balanced.

The effect of floating vegetation on denitrification and greenhouse gas production in wetland mesocosms

Science.gov (United States)

Jacobs, A. E.; Harrison, J. A.

2012-12-01

Anthropogenic intensification of nitrogen (N) loading to aquatic ecosystems is widespread and can lead to the degradation of these systems. Wetlands are important sites for N removal via denitrification, the microbially mediated reduction of reactive nitrate to inert N2 gas, but they can also produce high levels of greenhouse gases. Floating plants play an important role in encouraging denitrification, since they create low oxygen conditions that may favor denitrification. We investigated whether wetland sediments with floating plant cover had higher denitrification and greenhouse gas production rates than wetland sediments without floating plants. Replicate flow-through mesocosms with wetland sediment and water were constructed in a growth chamber to mimic the wetland where the sediment and water were collected. Mesocosm treatments were covered with floating vegetation (duckweed), an opaque tarp, or no cover to determine how cover type affects denitrification and greenhouse gas production and whether biotic or abiotic factors are likely responsible for observed differences. Denitrification and greenhouse gas production rates were calculated by measuring excess N2 gas, methane, and nitrous oxide concentrations in the water column and measuring the gas exchange rates between the water column and the atmosphere. Gas exchange rates were measured using an inert volatile tracer added to the water column and accumulation of gas in the mesocosm headspace. Additional mesocosm experiments were performed to determine how duckweed-dominated wetland systems respond to nitrogen loading and which mechanism for lowering dissolved oxygen concentrations is important in affecting denitrification under floating vegetation. Mesocosms with floating vegetation had lower dissolved oxygen than no cover or tarp-covered mesocosms, which is consistent with field and literature observations. Water flowing out of the mesocosms had statistically lower total nitrogen and nitrate concentrations

Effect of Temperature on Oxygen Profiles and Denitrification Rates in Freshwater Sediments

NARCIS (Netherlands)

Klein, de Jeroen J.M.; Overbeek, Ciska C.; Juncher Jørgensen, Christian

2017-01-01

Vegetated ditches and wetlands are important sites for nutrient removal in agricultural catchments. About half of the influx of inorganic nitrogen can be removed from these ecosystems by denitrification. Previous studies have shown that denitrification in aquatic ecosystems is strongly temperature

Transport zonation limits coupled nitrification-denitrification in permeable sediments

DEFF Research Database (Denmark)

Kessler, Adam John; Glud, R.N.; Cardenas, M.B.

2013-01-01

- and N-15-N-2 gas. The measured two-dimensional profiles correlate with computational model simulations, showing a deep pool of N-2 gas forming, and being advected to the surface below ripple peaks. Further isotope pairing calculations on these data indicate that coupled nitrification......-denitrification is severely limited in permeable sediments because the flow and transport field limits interaction between oxic and anoxic pore water. The approach allowed for new detailed insight into subsurface denitrification zones in complex permeable sediments....

Using rates of oxygen and nitrate reduction to map the subsurface distribution of groundwater denitrification

Science.gov (United States)

Kolbe, T.; De Dreuzy, J. R.; Abbott, B. W.; Aquilina, L.; Babey, T.; Green, C. T.; Fleckenstein, J. H.; Labasque, T.; Laverman, A.; Marçais, J.; Peiffer, S.; Thomas, Z.; Pinay, G.

2017-12-01

Widespread fertilizer application over the last 70 years has caused serious ecological and socioeconomic problems in aquatic and estuarine ecosystems. When surplus nitrogen leaches as nitrate (a major groundwater pollutant) to the aquifer, complex flow dynamics and naturally occurring degradation processes control its transport. Under the conditions of depleted oxygen and abundant electron donors, microorganisms reduce NO3- to N2 (denitrification). Denitrification rates vary over orders of magnitude among sites within the same aquifer, complicating estimation of denitrification capacity at the catchment scale. Because it is impractical or impossible to access the subsurface to directly quantify denitrification rates, reactivity is often assumed to occur continuous along flowlines, potentially resulting in substantial over- or underestimation of denitrification. Here we investigated denitrification in an unconfined crystalline aquifer in western France using a combination of common tracers (chlorofluorocarbons, O2, NO3-, and N2) measured in 16 wells to inform a time-based modeling approach. We found that spatially variable denitrification rates arise from the intersection of nitrate rich water with reactive zones defined by the abundance of electron donors (primarily pyrite). Furthermore, based on observed reaction rates of the sequential reduction of oxygen and nitrate, we present a general framework to estimate the location and intensity of the reactive zone in aquifers. Accounting for the vertical distribution of reaction rates results in large differences in estimations of net denitrification rates that assume homogeneous reactivity. This new framework provides a tractable approach for quantifying catchment and regional groundwater denitrification rates that could be used to improve estimation of groundwater resilience to nitrate pollution and develop more realistic management strategies.

Relationships Between Denitrifier Abundance, Denitrifier Diversity and Denitrification in Gulf of Mexico Hypoxic Zone Sediments.

Science.gov (United States)

Proctor, L. M.; Childs, C.; MacAuley, S.

2002-12-01

The largest zone of anthropogenic bottom water hypoxia in the Western Hemisphere occurs seasonally in the northern Gulf of Mexico. This hypoxic zone reaches its greatest extent in the summer months and is a consequence of seasonal stratification of the water column combined with the decomposition of organic matter derived from accelerated rates of primary production. The enhanced primary production is driven by inorganic nitrogen input from the Mississippi River and these conditions would seem ideal for supporting high levels of denitrification. Yet sediment denitrification exhibited a wide range, even at the height of the seasonal hypoxia. Therefore, we compared benthic denitrifier abundances and denitrifier diversity at several stations over two seasons exhibiting extremes in denitrification to evaluate the relationship between abundances, diversity and denitrification levels. Sediment denitrification ranged from 20 to 100 umol m-2 h-1, with rates in July, 2000 approximately half that observed in July, 2001. The highest rates were generally observed at stations with bottom water DO concentrations between 1 and 3 mg l-1. Relative denitrifier abundances, using nirS and nirK as proxies for denitrifiers, suggested a direct relationship between abundances and denitrification while denitrifier diversity, measured by T-RFLPs of nirS and nirK, suggested an inverse relationship between diversity and denitrification. These results suggest that several factors are important in understanding what controls denitrification in Gulf of Mexico hypoxic zone sediments.

Glacial-Holocene variability in pelagic denitrification and OMZ intensity along the NW Mexican Margin

Science.gov (United States)

Ontiveros Cuadras, J. F.; Thunell, R.; Ruiz-Fernandez, A. C.; Machain-Castillo, M. L.; Tappa, E.

2017-12-01

Denitrification of fixed nitrogen represents a substantial loss of bioavailable nitrogen from the ocean, thus playing a major role in the global nitrogen cycle. Water-column (pelagic) denitrification occurs mostly in the oxygen minimum zones (OMZs), which are situated beneath coastal upwelling areas that are characterized by high settling fluxes of organic detritus and high rates of oxygen utilization from remineralization. Our study uses biogenic components (total organic carbon and opal) and δ15N values of sediments from the NW Mexican Margin to reconstruct variations in denitrification and strength of the OMZ in the eastern tropical North Pacific (ETNP) for the last 36,000 years. During the last glacial period (LGM, 23-18 kyr) the associations between relatively low δ15N values (7-8‰) and low TOC (2-4%) and opal (1-4%) content indicates reduced denitrification due to reduced upwelling and decreased flux of organic matter through the OMZ. This was followed by abrupt acceleration of water-column denitrification (δ15N, 7-10‰) and the strengthening of the OMZ during the latter half of Heinrich Stadial 1 (HS1; 18-14.7 kyr). However, the biogenic component of sediments deposited during HS1 do not increase appreciably, suggesting that the increase in denitrification was not driven by an increase in productivity. Furthermore, the increase in δ15N precedes the deglacial decrease in planktonic foraminiferal δ18O which mostly occurs during the Bolling Alerod (14.7-12.9 kyr). This suggests that the increase in denitrification was not a response to surface warming. Rather, we attribute the rapid increase in denitrification during HS1 to reduced ventilation of the ETNP OMZ. Following the peak in denitrification at the end of HS1, we observe a small but steady decline in δ15N over the last 15 kyr. Higher TOC in Holocene sediments relative to glacial sediments suggests that increased productivity has played a role in maintaining a strong OMZ throughout the Holocene.

Irrigation of DOC-rich liquid promotes potential denitrification rate and decreases N

NARCIS (Netherlands)

Qin, Shuping; Hu, Chunsheng; Clough, Tim J.; Luo, Jiafa; Oenema, Oene; Zhou, Shungui

2017-01-01

Lack of dissolved organic carbon (DOC) is generally one of the key factors limiting denitrification in subsoil beneath the root zone. Despite a number of laboratory DOC amendment studies, the effects of in situ DOC infiltration on subsoil denitrification, and on subsequent end product

Descriptive and predictive assessment of enzyme activity and enzyme related processes in biorefinery using IR spectroscopy and chemometrics

DEFF Research Database (Denmark)

Baum, Andreas

the understanding of the structural properties of the extracted pectin. Secondly, enzyme kinetics of biomass converting enzymes was examined in terms of measuring enzyme activity by spectral evolution profiling utilizing FTIR. Chemometric multiway methods were used to analyze the tensor datasets enabling the second......-order calibration advantage (reference Theory of Analytical chemistry). As PAPER 3 illustrates the method is universally applicable without the need of any external standards and was exemplified by performing quantitative enzyme activity determinations for glucose oxidase, pectin lyase and a cellolytic enzyme blend...... (Celluclast 1.5L). In PAPER 4, the concept is extended to quantify enzyme activity of two simultaneously acting enzymes, namely pectin lyase and pectin methyl esterase. By doing so the multiway methods PARAFAC, TUCKER3 and NPLS were compared and evaluated towards accuracy and precision....

General discussion about enzymes activities of radiation injury

International Nuclear Information System (INIS)

Vucicevic, M.; Sukalo, I.

1989-01-01

Researching reliable and practical indicators of radiation injury, however, is very interesting and considerable department of scientific studies, practical and theoretical. Enzymes activities are among biochemical indicators which are changed after radiation injury. Activity of these specific proteins is important in regulation of every biochemical reaction in existing beings. Biological macromolecules can be damaged by radiation or the cell permeability can be changed. All of these influence directly on enzymes activities. In this paper we present the review of the all important enzymes, indicators of the radiation injury, which variances on reference to normal values are significant of the functional and the structural changes of essential organs (author)

General discussion about enzymes activities of radiation injury

Energy Technology Data Exchange (ETDEWEB)

Vucicevic, M; Sukalo, I [Institute of Nuclear Sciences Boris Kidric, Vinca, Beograd (Serbia and Montenegro)

1989-07-01

Researching reliable and practical indicators of radiation injury, however, is very interesting and considerable department of scientific studies, practical and theoretical. Enzymes activities are among biochemical indicators which are changed after radiation injury. Activity of these specific proteins is important in regulation of every biochemical reaction in existing beings. Biological macromolecules can be damaged by radiation or the cell permeability can be changed. All of these influence directly on enzymes activities. In this paper we present the review of the all important enzymes, indicators of the radiation injury, which variances on reference to normal values are significant of the functional and the structural changes of essential organs (author)

Actinomycete enzymes and activities involved in straw saccharification

Energy Technology Data Exchange (ETDEWEB)

McCarthy, A J; Ball, A S [Liverpool Univ. (UK). Dept. of Genetics and Microbiology

1990-01-01

This research programme has been directed towards the analysis of actinomycete enzyme systems involved in the degradation of plant biomass (lignocellulose.) The programme was innovative in that a novel source of enzymes was systematically screened and wheat straw saccharifying activity was the test criterion. Over 200 actinomycete strains representing a broad taxonomic range were screened. A range of specific enzyme activities were involved and included cellulase, xylanase, arabinofuranosidase, acetylesterase, {beta}-xylosidase and {beta}-glucosidase. Since hemicellulose (arabinoxylan) was the primary source of sugar, xylanases were characterized. The xylan-degrading systems of actinomycetes were complex and nonuniform, with up to six separate endoxylanases identified in active strains. Except for microbispora bispora, actinomycetes were found to be a poor source of cellulase activity. Evidence for activity against the lignin fraction of straw was produced for a range of actinomycete strains. While modification reactions were common, cleavage of inter-monomer bonds, and utilization of complex polyphenolic compounds were restricted to two strains: Thermomonospora mesophila and Streptomyces badius. Crude enzyme preparations from actinomycetes can be used to generate sugar, particularly pentoses, directly from cereal straw. The potential for improvements in yield rests with the formulation to cooperative enzyme combinations from different strains. The stability properties of enzymes from thermophilic strains and the general neutral to alkali pH optima offer advantages in certain process situations. Actinomycetes are a particularly rich source of xylanases for commercial application and can rapidly solubilise a lignocarbohydrate fraction of straw which may have both product and pretreatment potential. 31 refs., 4 figs., 5 tabs.

Denitrification in the Mississippi River network controlled by flow through river bedforms

Science.gov (United States)

Gomez-Velez, Jesus D.; Harvey, Judson W.; Cardenas, M. Bayani; Kiel, Brian

2015-01-01

Increasing nitrogen concentrations in the world’s major rivers have led to over-fertilization of sensitive downstream waters1, 2, 3, 4. Flow through channel bed and bank sediments acts to remove riverine nitrogen through microbe-mediated denitrification reactions5, 6, 7, 8, 9, 10. However, little is understood about where in the channel network this biophysical process is most efficient, why certain channels are more effective nitrogen reactors, and how management practices can enhance the removal of nitrogen in regions where water circulates through sediment and mixes with groundwater - hyporheic zones8, 11, 12. Here we present numerical simulations of hyporheic flow and denitrification throughout the Mississippi River network using a hydrogeomorphic model. We find that vertical exchange with sediments beneath the riverbed in hyporheic zones, driven by submerged bedforms, has denitrification potential that far exceeds lateral hyporheic exchange with sediments alongside river channels, driven by river bars and meandering banks. We propose that geomorphic differences along river corridors can explain why denitrification efficiency varies between basins in the Mississippi River network. Our findings suggest that promoting the development of permeable bedforms at the streambed - and thus vertical hyporheic exchange - would be more effective at enhancing river denitrification in large river basins than promoting lateral exchange through induced channel meandering. 

Modeling Nitrous Oxide Production during Biological Nitrogen Removal via Nitrification and Denitrification: Extensions to the General ASM Models

DEFF Research Database (Denmark)

Ni, Bing-Jie; Ruscalleda, Maël; Pellicer i Nàcher, Carles

2011-01-01

on N2O production from four different mixed culture nitrification and denitrification reactor study reports. Modeling results confirm that hydroxylamine oxidation by ammonium oxidizers (AOB) occurs 10 times slower when NO2– participates as final electron acceptor compared to the oxic pathway. Among......Nitrous oxide (N2O) can be formed during biological nitrogen (N) removal processes. In this work, a mathematical model is developed that describes N2O production and consumption during activated sludge nitrification and denitrification. The well-known ASM process models are extended to capture N2O...

Nitrogen removal from sludge digester liquids by nitrification/denitrification or partial nitritation/anammox: environmental and economical considerations.

Science.gov (United States)

Fux, C; Siegrist, H

2004-01-01

In wastewater treatment plants with anaerobic sludge digestion, 15-20% of the nitrogen load is recirculated to the main stream with the return liquors from dewatering. Separate treatment of this ammonium-rich digester supernatant significantly reduces the nitrogen load of the activated sludge system. Two biological applications are considered for nitrogen elimination: (i) classical autotrophic nitrification/heterotrophic denitrification and (ii) partial nitritation/autotrophic anaerobic ammonium oxidation (anammox). With both applications 85-90% nitrogen removal can be achieved, but there are considerable differences in terms of sustainability and costs. The final gaseous products for heterotrophic denitrification are generally not measured and are assumed to be nitrogen gas (N2). However, significant nitrous oxide (N2O) production can occur at elevated nitrite concentrations in the reactor. Denitrification via nitrite instead of nitrate has been promoted in recent years in order to reduce the oxygen and the organic carbon requirements. Obviously this "achievement" turns out to be rather disadvantageous from an overall environmental point of view. On the other hand no unfavorable intermediates are emitted during anaerobic ammonium oxidation. A cost estimate for both applications demonstrates that partial nitritation/anammox is also more economical than classical nitrification/denitrification. Therefore autotrophic nitrogen elimination should be used in future to treat ammonium-rich sludge liquors.

Thermodynamic Activity-Based Progress Curve Analysis in Enzyme Kinetics.

Science.gov (United States)

Pleiss, Jürgen

2018-03-01

Macrokinetic Michaelis-Menten models based on thermodynamic activity provide insights into enzyme kinetics because they separate substrate-enzyme from substrate-solvent interactions. Kinetic parameters are estimated from experimental progress curves of enzyme-catalyzed reactions. Three pitfalls are discussed: deviations between thermodynamic and concentration-based models, product effects on the substrate activity coefficient, and product inhibition. Copyright © 2017 Elsevier Ltd. All rights reserved.

[Denitrification study of Elodea nuttallii-nitrogen cycling bacteria restoration in Meiliang Bay, Taihu Lake].

Science.gov (United States)

Zhao, Lin; Li, Zheng-Kui; Zhou, Tao; Wu, Ning-Mei; Ye, Zhong-Xiang; Liu, Dan-Dan

2013-08-01

Undisturbed sediment cores were collected from Meiliang Bay, Taihu Lake, and the integrated Elodea nuttallii-nitrogen cycling bacteria technology was applied as a restoration method. The effects of the Elodea nuttallii-nitrogen cycling bacteria technology on sediment denitrification was observed by isotope pairing technique. The highest denitrification rate of 104.64 micromol x (m2 x h)(-1) was achieved in sediments with Elodea nuttallii-nitrogen cycling bacteria assemblage. The abundance of nirS, nirK and nosZ genes involved in denitrification processes in the sediments (within 2 cm below the water-sediment interface) were measured by real-time quantitative PCR (RT-qPCR). The abundance of nirS and nosZ genes in the sediments with restoration treatments was increased, which was more than one order of magnitudes higher than that in bare sediments. The results indicated that the presence of macrophyte and nitrogen cycling bacteria could increase benthic nitrogen removal by facilitating coupled nitrification-denitrification and uncoupled nitrification-denitrification.

Denitrification in the water column of the central Baltic Sea

DEFF Research Database (Denmark)

Dalsgaard, Tage; De Brabandere, Loreto; Hall, Per

2013-01-01

denitrification ranged from 0.24 to 15.9 nM N2 h−1. Assuming that these rates were valid throughout the anoxic View the MathML source containing zone, depth integrated in situ denitrification rates of 0.06–2.11 mmol N m−2 d−1 were estimated. The thickness of this zone was generally 3–6 m, which is probably what...... View the MathML source or sulfide concentrations were converted to in situ rates using the measured water column concentrations of View the MathML source and sulfide and the actual measured relations between View the MathML source and sulfide concentrations and denitrification rates. In situ...... can be maintained through regular turbulent mixing induced by internal waves at the oxic–anoxic interface. However, layers of up to 55 m thickness with low O2 water (Math...

Nitrogen removal and electricity production at a double-chamber microbial fuel cell with cathode nitrite denitrification.

Science.gov (United States)

Yu, Yangyang; Zhao, Jianqiang; Wang, Sha; Zhao, Huimin; Ding, Xiaoqian; Gao, Kun

2017-12-01

Double-chamber microbial fuel cell was applied to investigate the performance of the electricity production and nitrite denitrification through feeding nitrite into the cathode. Factors influencing denitrification performance and power production, such as external resistance, influent nitrite concentration and Nitrite Oxygen Bacteria inhibitors, were studied. The results show that when the concentration of nitrite nitrogen and external resistance were 100 mg L -1 and 10 Ω, respectively, the nitrite denitrification reached the best state. The NaN 3 can inhibit nitrite oxidation effectively; meanwhile, the nitrite denitrification with N 2 O as the final products was largely improved. The [Formula: see text] was reduced to [Formula: see text], causing the cathode denitrification coulombic efficiency to exceed 100%. In chemoautotrophic bio-nitrification, microorganisms may utilize H 2 O to oxidize nitrite under anaerobic conditions. Proteobacteria might play a major role in the process of denitrification in MFC.

Immobilized enzyme reactor chromatography: Optimization of protein retention and enzyme activity in monolithic silica stationary phases

International Nuclear Information System (INIS)

Besanger, Travis R.; Hodgson, Richard J.; Green, James R.A.; Brennan, John D.

2006-01-01

Our group recently reported on the application of protein-doped monolithic silica columns for immobilized enzyme reactor chromatography, which allowed screening of enzyme inhibitors present in mixtures using mass spectrometry for detection. The enzyme was immobilized by entrapment within a bimodal meso/macroporous silica material prepared by a biocompatible sol-gel processing route. While such columns proved to be useful for applications such as screening of protein-ligand interactions, significant amounts of entrapped proteins leached from the columns owing to the high proportion of macropores within the materials. Herein, we describe a detailed study of factors affecting the morphology of protein-doped bioaffinity columns and demonstrate that specific pH values and concentrations of poly(ethylene glycol) can be used to prepare essentially mesoporous columns that retain over 80% of initially loaded enzyme in an active and accessible form and yet still retain sufficient porosity to allow pressure-driven flow in the low μL/min range. Using the enzyme γ-glutamyl transpeptidase (γ-GT), we further evaluated the catalytic constants of the enzyme entrapped in capillary columns with different silica morphologies as a function of flowrate and backpressure using the enzyme reactor assay mode. It was found that the apparent activity of the enzyme was highest in mesoporous columns that retained high levels of enzyme. In such columns, enzyme activity increased by ∼2-fold with increases in both flowrate (from 250 to 1000 nL/min) and backpressure generated (from 500 to 2100 psi) during the chromatographic activity assay owing to increases in k cat and decreases in K M , switching from diffusion controlled to reaction controlled conditions at ca. 2000 psi. These results suggest that columns with minimal macropore volumes (<5%) are advantageous for the entrapment of soluble proteins for bioaffinity and bioreactor chromatography

The role of nitrifier denitrification in the production of nitrous oxide revisited

NARCIS (Netherlands)

Wrage-Mönnig, Nicole; Horn, Marcus A.; Well, Reinhard; Müller, Christoph; Velthof, Gerard; Oenema, Oene

2018-01-01

Nitrifier denitrification is the reduction of nitrite (NO2 −) by ammonia-oxidizing bacteria. This process may account for up to 100% of nitrous oxide (N2O) emissions from ammonium (NH4 +) in soils and is more significant than classical denitrification under some conditions. Investigations of

High-Resolution Denitrification Kinetics in Pasture Soils Link N2O Emissions to pH, and Denitrification to C Mineralization.

Directory of Open Access Journals (Sweden)

Md Sainur Samad

Full Text Available Denitrification in pasture soils is mediated by microbial and physicochemical processes leading to nitrogen loss through the emission of N2O and N2. It is known that N2O reduction to N2 is impaired by low soil pH yet controversy remains as inconsistent use of soil pH measurement methods by researchers, and differences in analytical methods between studies, undermine direct comparison of results. In addition, the link between denitrification and N2O emissions in response to carbon (C mineralization and pH in different pasture soils is still not well described. We hypothesized that potential denitrification rate and aerobic respiration rate would be positively associated with soils. This relationship was predicted to be more robust when a high resolution analysis is performed as opposed to a single time point comparison. We tested this by characterizing 13 different temperate pasture soils from northern and southern hemispheres sites (Ireland and New Zealand using a fully automated-high-resolution GC detection system that allowed us to detect a wide range of gas emissions simultaneously. We also compared the impact of using different extractants for determining pH on our conclusions. In all pH measurements, soil pH was strongly and negatively associated with both N2O production index (IN2O and N2O/(N2O+N2 product ratio. Furthermore, emission kinetics across all soils revealed that the denitrification rates under anoxic conditions (NO+N2O+N2 μmol N/h/vial were significantly associated with C mineralization (CO2 μmol/h/vial measured both under oxic (r2 = 0.62, p = 0.0015 and anoxic (r2 = 0.89, p<0.0001 conditions.

Modification of polymer surfaces to enhance enzyme activity and stability

DEFF Research Database (Denmark)

Hoffmann, Christian

Enzyme immobilization is an important concept for the development of improved biocatalytic processes, primarily through facilitated separation procedures. However, enzyme immobilization usually comes at a price of reduced biocatalytic activity. For this reason, different immobilization methods have...... already been developed, combining the same goal to improve enzyme activity, stability and selectivity. Polymer materials have shown, due to their easy processibility and versatile properties, high potential as enzyme support. However, in order to achieve improved enzyme performance, the combination...... on their tailored surface modification in order to obtain improved enzyme-support systems. Firstly, an off-stoichiometric thiol-ene (OSTE) thermosetting material was used for the development of a screening platform allowing the investigation of micro-environmental effects and their impact on the activity...

Orbital-scale denitrification changes in the Eastern Arabian Sea during the last 800 kyrs.

Science.gov (United States)

Kim, Ji-Eun; Khim, Boo-Keun; Ikehara, Minoru; Lee, Jongmin

2018-05-04

Denitrification in the Arabian Sea is closely related to the monsoon-induced upwelling and subsequent phytoplankton production in the surface water. The δ 15 N values of bulk sediments collected at Site U1456 of the International Ocean Discovery Program (IODP) Expedition 355 reveal the orbital-scale denitrification history in response to the Indian Monsoon. Age reconstruction based on the correlation of planktonic foraminifera (Globigerinoides ruber) δ 18 O values with the LR04 stack together with the shipboard biostratigraphic and paleomagnetic data assigns the study interval to be 1.2 Ma. Comparison of δ 15 N values during the last 800 kyrs between Site U1456 (Eastern Arabian Sea) and Site 722B (Western Arabian Sea) showed that δ 15 N values were high during interglacial periods, indicating intensified denitrification, while the opposite was observed during glacial periods. Taking 6‰ as the empirical threshold of denitrification, the Eastern Arabian Sea has experienced a persistent oxygen minimum zone (OMZ) to maintain strong denitrification whereas the Western Arabian Sea has undergone OMZ breakdown during some glacial periods. The results of this study also suggests that five principal oceanographic conditions were changed in response to the Indian Monsoon following the interglacial and glacial cycles, which controls the degree of denitrification in the Arabian Sea.

Thermal hydrolysis of sludge and the use of hydrolysate as carbon source for denitrification

Energy Technology Data Exchange (ETDEWEB)

Barlindhaug, J

1995-10-01

As a consequence of the North Sea- and the Baltic Sea Treaties as well as the Wastewater Directive of the EU, several large wastewater treatment plants discharging to sensitive receiving waters have to include phosphorus as well as nitrogen removal. This thesis evaluates the so called NTH-process for nutrient removal. In this process pre-precipitation is used in front of a biological nitrogen removal step that is based on a combination of pre- and post-denitrification in moving bed biofilm reactors. The biological step is followed by a final separation step, possibly after coagulant addition. Carbon source for the post denitrification step is made available by hydrolysis of the sludge produced. The idea is that the particulate organic matter, which in a traditional pre-denitrification step would have to be enzymatically hydrolyzed, can be more efficiently hydrolyzed in a concentrated sidestream and used in a post-denitrification step. In the thesis hydrolyzed sludge is used as a carbon source for denitrification. The objective is to investigate the influence of varying hydrolysis conditions on the composition and amount of the thermal hydrolysate produced, as well as the quality of the hydrolysate as a carbon source for denitrification. 201 refs., 78 refs., 53 tabs.

Impact of chloride on denitrification potential in roadside wetlands.

Science.gov (United States)

Lancaster, Nakita A; Bushey, Joseph T; Tobias, Craig R; Song, Bongkeun; Vadas, Timothy M

2016-05-01

Developed landscapes are exposed to changes in hydrology and water chemistry that limit their ability to mitigate detrimental impacts to coastal water bodies, particularly those that result from stormwater runoff. The elevated level of impervious cover increases not only runoff but also contaminant loading of nutrients, metals, and road salt used for deicing to water bodies. Here we investigate the impact that road salt has on denitrification in roadside environments. Sediments were collected from a series of forested and roadside wetlands and acclimated with a range of Cl(-) concentrations from 0 to 5000 mg L(-1) for 96 h. Denitrification rates were measured by the isotope pairing technique using (15)N-NO3(-), while denitrifying community structures were compared using terminal restriction fragment length polymorphism (T-RFLP) of nitrous oxide reductase genes (nosZ). Chloride significantly (p wetlands at a Cl(-) dosage of 2500 or 5000 mg L(-1), but the decrease in denitrification rates was less and not significant for the roadside wetlands historically exposed to elevated concentrations of Cl(-). The difference could not be attributed to other significant changes in conditions, such as DOC concentrations, N species concentrations, or pH levels. Denitrifying communities, as measured by T-RFs of the nosZ gene, in the roadside wetlands with elevated concentration of Cl(-) were distinctly different and more diverse compared to forested wetlands, and also different in roadside wetlands after 96 h exposures to Cl(-). The shifts in denitrifying communities seem to minimize the decrease in denitrification rates in the wetlands previously exposed to Cl. As development results in more Cl(-) use and exposure to a broad range of natural or manmade wetland structures, an understanding of the seasonal effect of Cl on denitrification processes in these systems would aid in design or mitigation of the effects on N removal rates. Copyright © 2016 Elsevier Ltd. All rights

Environmental and Microbial Features Affecting Denitrification and Anammox Hotspots in an Estuarine Ecosystem

Science.gov (United States)

Lisa, J.; Song, B.; Lefcheck, J. S.; Tobias, C. R.

2016-02-01

Biogeochemical hotspots are characterized as a few sites that exhibit extremely high reaction rates relative to surrounding area, and often account for a high percentage of the overall reaction rates in an ecosystem. Criteria for quantitatively identifying these sites have not been well established. Further, the underlying mechanisms of hotspots have been described in terms of environmental conditions, with little attention paid to the microbial community. The objectives of this study were to establish quantitative criteria to identify denitrification and anammox hotspots, and determine the underlying microbial and environmental factors responsible for elevated N2 production. We used 15N isotope pairing incubation experiments to measure denitrification and anammox rates in the New River Estuary, NC. Quantitative PCR assays of nitrous oxide reductase (nosZ Clades I and II) and hydrazine oxidoreductase (hzo) genes were conducted to estimate denitrifier and anammox abundance. Structural Equation Modeling (SEM) was used to elucidate complex causal relationships between environmental and biological variables. Denitrification hotspots, quantitatively defined as statistical outliers, accounted for 35.6% total denitrification while comprising only 7.3% of the sites. Anammox hotspots,10.6% of the sites, accounted for 60.9% of total anammox. SEM revealed increased sediment organics at lower salinities supported higher functional gene abundance, which in turn resulted in higher N2 production. Surprisingly, denitrification rates were significantly and positively correlated with nosZ Clade II gene abundance, after accounting for the non-significant contributions of the naturally more abundant nosZ Clade I, and other environmental covariates. This is the first time that a quantitative definition of biogeochemical hotspots was put forth and used to determine the importance of anammox and denitrification hotspots in estuarine nitrogen removal capacity. Despite the low area

Comparison of heterotrophic and autotrophic denitrification processes for nitrate removal from phosphorus-limited surface water.

Science.gov (United States)

Wang, Zheng; He, Shengbing; Huang, Jungchen; Zhou, Weili; Chen, Wanning

2018-03-29

Phosphorus (P) limitation has been demonstrated for micro-polluted surface water denitrification treatment in previous study. In this paper, a lab-scale comparative study of autotrophic denitrification (ADN) and heterotrophic denitrification (HDN) in phosphorus-limited surface water was investigated, aiming to find out the optimal nitrogen/phosphorus (N/P) ratio and the mechanism of the effect of P limitation on ADN and HDN. Furthermore, the optimal denitrification process was applied to the West Lake denitrification project, aiming to improve the water quality of the West Lake from worse than grade V to grade IV (GB3838-2006). The lab-scale study showed that the lack of P indeed inhibited HDN more greatly than ADN. The optimal N/P ratio for ADN and HDN was 25 and a 0.15 mg PO 4 3- -P L -1 of microbial available phosphorus (MAP) was observed. P additions could greatly enhance the resistance of ADN and HDN to hydraulic loading shock. Besides, The P addition could effectively stimulate the HDN performance via enriching the heterotrophic denitrifiers and the denitrifying phosphate-accumulating organisms (DNPAOs). Additionally, HDN was more effective and cost-effective than ADN for treating P-limited surface water. The study of the full-scale HDBF (heterotrophic denitrification biofilter) indicated that the denitrification performance was periodically impacted by P limitation, particularly at low water temperatures. Copyright © 2018 Elsevier Ltd. All rights reserved.

Isotopologue fractionation during N(2)O production by fungal denitrification.

Science.gov (United States)

Sutka, Robin L; Adams, Gerard C; Ostrom, Nathaniel E; Ostrom, Peggy H

2008-12-01

Identifying the importance of fungi to nitrous oxide (N2O) production requires a non-intrusive method for differentiating between fungal and bacterial N2O production such as natural abundance stable isotopes. We compare the isotopologue composition of N2O produced during nitrite reduction by the fungal denitrifiers Fusarium oxysporum and Cylindrocarpon tonkinense with published data for N2O production during bacterial nitrification and denitrification. The fractionation factors for bulk nitrogen isotope values for fungal denitrification were in the range -74.7 to -6.6 per thousand. There was an inverse relationship between the absolute value of the fractionation factors and the reaction rate constant. We interpret this in terms of variation in the relative importance of the rate constants for diffusion and enzymatic reduction in controlling the net isotope effect for N2O production during fungal denitrification. Over the course of nitrite reduction, the delta(18)O values for N2O remained constant and did not exhibit a relationship with the concentration characteristic of an isotope effect. This probably reflects isotopic exchange with water. Similar to the delta(18)O data, the site preference (SP; the difference in delta(15)N between the central and outer N atoms in N2O) was unrelated to concentration during nitrite reduction and, therefore, has the potential to act as a conservative tracer of production from fungal denitrification. The SP values of N2O produced by F. oxysporum and C. tonkinense were 37.1 +/- 2.5 per thousand and 36.9 +/- 2.8 per thousand, respectively. These SP values are similar to those obtained in pure culture studies of bacterial nitrification but quite distinct from SP values for bacterial denitrification. The large magnitude of the bulk nitrogen isotope fractionation and the delta(18)O values associated with fungal denitrification are distinct from bacterial production pathways; thus multiple isotopologue data holds much promise for

ENZYME ACTIVITIES OF PADDY SOILS AND RELATIONSHIPS WITH THE SOIL PROPERTIES

Directory of Open Access Journals (Sweden)

Rıdvan KIZILKAYA

1998-03-01

Full Text Available This study was carried out to determine the effect of soil properties on enzyme activities of paddy soils, the sample of which were taken from Üçpınar, Harız, Doğancı, Kaygusuz, Emenli, Sarıköy and Gelemenağarı villages where rice cultivation is an intensive agricultural system. In this study, soil properties having effects on urease, phosphatase, ß-glucosidase and catalase enzyme activities were setforth. Urease enzyme activities of soil samples varied from 24.12 to 39.03 mg N 100 g dry soil -1 . Significant correlations were determined between urease enzyme activities and organic matter (r = 0.89**, extractable Mn (r = 0.74**, exchangable K (r = 0.73** and total P content of soil (r = 0.81*. Acid phosphatase enzyme activity varied between 3.00-17.44 mg phenol 100 g dry soil -1 , alkaline phosphatase enzyme activity between 12.00-25.53 mg phenol 100 g dry soil-1 . Exchangable Mg (r = 0.71* and extractable Cu (r = 0.74* were found to have positive effect on acid phosphatase enzyme activity and pH (r = 0.73*, exchangable Ca (r = 0.74*, exchangable Mg (r = 0.71*, exchangable total basic cations (r = 0.79* and extractable Cu (r = 0.70* had positive effects on alkaline phosphatase enzyme activity, whereas total P (r = - 0.84** affected the activity negatively. ß-glucosidase enzyme activity was measured to vary between 1.12-3.64 mg salingen 100 g dry soil -1 . It was also observed that extractable Zn content of soil samples (r = - 0.97** had negative effect on ß-glucosidase activity, wheras total exchangable acidic cations (r = 0.70* affected the activity positively. Catalase enzyme activities of soils changed between 5.25 - 9.00 mg O2 5 g dry soil -1 . Significant correlations were found between catalase activities and fraction of soils and extractable Fe content. Positive correlations, however, were determined between catalase activities and clay fraction (r = 0.82* and salt content (r = 0.83** of samples.

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

Science.gov (United States)

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

2017-04-01

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

Nitrogen speciation and trends, and prediction of denitrification extent, in shallow US groundwater

Science.gov (United States)

Hinkle, Stephen R.; Tesoriero, Anthony J.

2014-01-01

Uncertainties surrounding nitrogen cycling complicate assessments of the environmental effects of nitrogen use and our understanding of the global carbon–nitrogen cycle. In this paper, we synthesize data from 877 ambient-monitoring wells across the US to frame broad patterns of nitrogen speciation and trends. At these sites, groundwater frequently contains substantial co-occurring NO3− and XSN2 (N2 from denitrification), reflecting active/ongoing denitrification and/or a mixture of undenitrified and denitrified groundwater. NO3− and NH4+ essentially do not co-occur, indicating that the dominant source of NH4+ at these sites likely is not dissimilatory reduction of NO3− to NH4+. Positive correlations of NH4+ with apparent age, CH4, dissolved organic carbon, and indicators of reduced conditions are consistent with NH4+ mobilization from degradation of aquifer organic matter and contraindicate an anthropogenic source of NH4+ for most sites. Glacial aquifers and eastern sand and gravel aquifers generally have lower proportions of NO3− and greater proportions of XSN2 than do fractured rock and karst aquifers and western sand and gravel aquifers. NO3− dominates in the youngest groundwater, but XSN2 increases as residence time increases. Temporal patterns of nitrogen speciation and concentration reflect (1) changing NO3− loads over time, (2) groundwater residence-time controls on NH4+ mobilization from solid phases, and (3) groundwater residence-time controls on denitrification. A simple classification tree using readily available variables (a national coverage of soil water depth, generalized geology) or variables reasonably estimated in many aquifers (residence time) identifies categorical denitrification extent (50%) with 79% accuracy in an independent testing set, demonstrating a predictive application based on the interconnected effects of redox, geology, and residence time.

Controlled Carbon Source Addition to an Alternating Nitrification-Denitrification Wastewater Treatment Process Including Biological P Removal

DEFF Research Database (Denmark)

Isaacs, Steven Howard; Henze, Mogens

1995-01-01

The paper investigates the effect of adding an external carbon source on the rate of denitrification in an alternating activated sludge process including biological P removal. Two carbon sources were examined, acetate and hydrolysate derived from biologically hydrolyzed sludge. Preliminary batch ...

Field tracer test for denitrification in a pyrite-bearing schist aquifer

International Nuclear Information System (INIS)

Pauwels, H.; Kloppmann, W.; Foucher, J.-C.; Martelat, A.; Fritsche, V.

1998-01-01

A small-scale artificial tracer test performed on a schist aquifer in Brittany has helped clarify mechanisms and kinetics of in situ autotrophic denitrification. NO 3 was injected as a pulse simultaneously with a conservative tracer - Br - . During the test, which lasted 210 h, 73% of the injected Br - was recovered, as against only 47% of the NO 3 . The 26% difference in the recovery of the two injected species is interpreted as being the result of denitrification, in part due to the direct oxidation of pyrite present in the solid aquifer according to the reaction: 5FeS 2 +14NO 3 - +4H + approaches7N 2 +10SO 4 2- +5Fe 2+ +2H 2 O, and in part due to subsequent iron oxidation according to the reaction: NO 3 - +5Fe 2+ +6H + approaches1/2N 2 +5Fe 3+ +3H 2 O. Despite the potential increase in SO 4 and Fe resulting from denitrification through pyrite oxidation, the concentrations of these elements in the groundwater remain moderate due to the precipitation of minerals such as jarosite and/or natroalunite. Tracer transfer takes place in a heterogeneous medium which, according to the breakthrough curves, can be simplified to a dual-porosity aquifer comprising a high-permeability (fractures or large fissures) medium of low porosity from which only minor denitrification of circulating NO 3 -bearing water was observed and a low-permeability (small fissures) medium of high porosity which induces a higher denitrification rate in the circulating NO 3 -bearing water. The kinetics of the denitrification reaction are high compared with results obtained for other environments and can be described by a first-order model with a half life of 7.9 days for the low-porosity medium and only 2.1 days for the high-porosity medium. (Copyright (c) 1998 Elsevier Science B.V., Amsterdam. All rights reserved.)

Microbial Enzyme Activity and Carbon Cycling in Grassland Soil Fractions

Science.gov (United States)

Allison, S. D.; Jastrow, J. D.

2004-12-01

Extracellular enzymes are necessary to degrade complex organic compounds present in soils. Using physical fractionation procedures, we tested whether old soil carbon is spatially isolated from degradative enzymes across a prairie restoration chronosequence in Illinois, USA. We found that carbon-degrading enzymes were abundant in all soil fractions, including macroaggregates, microaggregates, and the clay fraction, which contains carbon with a mean residence time of ~200 years. The activities of two cellulose-degrading enzymes and a chitin-degrading enzyme were 2-10 times greater in organic matter fractions than in bulk soil, consistent with the rapid turnover of these fractions. Polyphenol oxidase activity was 3 times greater in the clay fraction than in the bulk soil, despite very slow carbon turnover in this fraction. Changes in enzyme activity across the restoration chronosequence were small once adjusted for increases in soil carbon concentration, although polyphenol oxidase activity per unit carbon declined by 50% in native prairie versus cultivated soil. These results are consistent with a `two-pool' model of enzyme and carbon turnover in grassland soils. In light organic matter fractions, enzyme production and carbon turnover both occur rapidly. However, in mineral-dominated fractions, both enzymes and their carbon substrates are immobilized on mineral surfaces, leading to slow turnover. Soil carbon accumulation in the clay fraction and across the prairie restoration chronosequence probably reflects increasing physical isolation of enzymes and substrates on the molecular scale, rather than the micron to millimeter scale.

Performance evaluation of pilot scale sulfur-oxidizing denitrification for treatment of metal plating wastewater.

Science.gov (United States)

Flores, Angel S P; Gwon, Eun-Mi; Sim, Dong-Min; Nisola, Grace; Galera, Melvin M; Chon, Seung-Se; Chung, Wook-Jin; Pak, Dae-Won; Ahn, Zou Sam

2006-01-01

A full-scale and two pilot-scale upflow sulfur-oxidizing denitrification (SOD) columns were evaluated using metal plating wastewater as feed. The sludge was autotrophically enriched, and inoculated in the SOD columns attached to the effluent line of three metal plating wastewater treatment facilities. The effects of activated carbon and aeration were also studied, and found effective for the removal of suspended solids and ammonia, respectively. The results showed that the constituents, such as the total nitrogen, nitrates, nitrites, ammonia, chemical oxygen demand (COD), and heavy metals, were effectively removed. The pH was observed to be maintained at 7-8 due to the alkalinity supplied by the sulfur-calcium carbonate (SC) pellet. The denitrification efficiency and start-up period were observed to be affected by the influent quality. Chromium, iron, nickel, copper, and zinc--the major heavy metal components of the influent--were effectively reduced at certain concentrations. Other metal ions were also detected and reduced to undetectable concentrations, but no trends in the comparison with denitrification were observed. From the results it can be concluded that SOD is effective for the removal of nitrogen, particularly nitrates, without a drastic pH change, and can effectively remove minute concentrations of heavy metals and COD in metal plating wastewaters.

Denitrification of nitrate waste solutions

International Nuclear Information System (INIS)

Bertolami, R.J.; Chao, E.I.; Choi, W.M.; Johnson, B.R.; Varlet, J.L.P.

1976-01-01

Growth rates for the denitrifying bacteria Pseudomonas Stutzeri were studied to minimize the time necessary to start up a bacterial denitrification reactor. Batch experiments were performed in nine 250-ml Erlenmeyer flasks, a 7-liter fermentor, and a 67-liter fermentor. All reactors maintained an anaerobic environment. Initial microorganism inoculum concentration was varied over four orders of magnitude. Initial nitrate and substrate carbon concentrations were varied from 200 to 6000 ppm and from 56 to 1596 ppm, respectively, with a carbon-to-nitrogen weight ratio of 1.18. In all experiments, except those with the highest initial substrate-to-bacteria ratio, no growth was observed due to substrate depletion during the lag period. In those experiments which did exhibit an increase in bacterial population, growth also stopped due to substrate depletion. A model simulating microbe growth during the induction period was developed, but insufficient data were available to properly adjust the model constants. Because of this, the model does not accurately predict microbe growth. The metabolism of Pseudomonas Stutzeri was studied in detail. This resulted in a prediction of the denitrification stoichiometry during steady state reactor operation. Iron was found to be an important component for bacterial anabolism

An additional simple denitrification bioreactor using packed gel envelopes applicable to industrial wastewater treatment.

Science.gov (United States)

Morita, Masahiko; Uemoto, Hiroaki; Watanabe, Atsushi

2007-08-15

A simple denitrification bioreactor for nitrate-containing wastewater without organic compounds was developed. This bioreactor consisted of packed gel envelopes in a single tank. Each envelope comprised two plates of gels containing Paracoccus denitrificans cells with an internal space between the plates. As an electron donor for denitrification, ethanol was injected into the internal space and not directly into the wastewater. P. denitrificans cells in the gel reduced nitrate to nitrogen gas by using the injected ethanol. Nitrate-containing desulfurization wastewater derived from a coal-fired thermal power plant was continuously treated with 20 packed gel envelopes (size, 1,000 x 900 x 12 mm; surface area, 1.44 m(2)) in a reactor tank (volume 1.5 m(3)). When the total nitrogen concentration in the inflow was around 150 mg-N x L(-1), the envelopes removed approximately 60-80% of the total nitrogen, and the maximum nitrogen removal rate was 5.0 g-N x day(-1) per square meter of the gel surface. This value corresponded to the volumetric nitrogen removal performance of 0.109 kg-N x m(-3) x day(-1). In each envelope, a high utilization efficiency of the electron donor was attained, although more than the double amount of the electron donor was empirically injected in the present activated sludge system to achieve denitrification when compared with the theoretical value. The bioreactor using the envelopes would be extremely effective as an additional denitrification system because these envelopes can be easily installed in the vacant spaces of preinstalled water treatment systems, without requiring additional facilities for removing surplus ethanol and sludge. (c) 2007 Wiley Periodicals, Inc.

Enzyme hydration, activity and flexibility : A neutron scattering approach

International Nuclear Information System (INIS)

Kurkal-Siebert, V.; Finney, J.L.; Daniel, R.M.; Smith, Jeremy C.

2006-01-01

Recent measurements have demonstrated enzyme activity at hydrations as low as 3%. The question of whether the hydration-induced enzyme flexibility is important for activity is addressed by performing picosecond dynamic neutron scattering experiments on pig liver esterase powders at various temperatures as well as solutions. At all temperatures and hydrations investigated here, significant quasielastic scattering intensity is found in the protein, indicating the presence of anharmonic, diffusive motion. As the hydration increases a temperature-dependent dynamical transition appears and strengthens involving additional diffusive motion. At low temperature, increasing hydration resulted in lower flexibility of the enzyme. At higher temperatures, systems containing sufficient number of water molecules interacting with the protein exhibit increased flexibility. The implication of these results is that, although the additional hydration-induced diffusive motion and flexibility at high temperatures in the enzyme detected here may be related to increased activity, they are not required for the enzyme to function

Predicting the denitrification capacity of sandy aquifers from shorter-term incubation experiments and sediment properties

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W. Eschenbach

2013-02-01

Full Text Available Knowledge about the spatial variability of denitrification rates and the lifetime of denitrification in nitrate-contaminated aquifers is crucial to predict the development of groundwater quality. Therefore, regression models were derived to estimate the measured cumulative denitrification of aquifer sediments after one year of incubation from initial denitrification rates and several sediment parameters, namely total sulphur, total organic carbon, extractable sulphate, extractable dissolved organic carbon, hot water soluble organic carbon and potassium permanganate labile organic carbon.

For this purpose, we incubated aquifer material from two sandy Pleistocene aquifers in Northern Germany under anaerobic conditions in the laboratory using the ¹⁵N tracer technique. The measured amount of denitrification ranged from 0.19 to 56.2 mg N kg⁻¹ yr⁻¹. The laboratory incubations exhibited high differences between non-sulphidic and sulphidic aquifer material in both aquifers with respect to all investigated sediment parameters. Denitrification rates and the estimated lifetime of denitrification were higher in the sulphidic samples. For these samples, the cumulative denitrification measured during one year of incubation (D_cum(365 exhibited distinct linear regressions with the stock of reduced compounds in the investigated aquifer samples. D_cum(365 was predictable from sediment variables within a range of uncertainty of 0.5 to 2 (calculated D_cum(365/measured D_cum(365 for aquifer material with a D_cum(365 > 20 mg N kg⁻¹ yr⁻¹. Predictions were poor for samples with lower D_cum(365, such as samples from the NO₃⁻ bearing groundwater zone, which includes the non-sulphidic samples, from the upper part of both aquifers where denitrification is not sufficient to

ANAEROBIC DIGESTION AND THE DENITRIFICATION IN UASB REACTOR

Directory of Open Access Journals (Sweden)

José Tavares de Sousa

2008-01-01

Full Text Available The environmental conditions in Brazil have been contributing to the development of anaerobic systems in the treatment of wastewaters, especially UASB - Upflow Anaerobic Sludge Blanket reactors. The classic biological process for removal of nutrients uses three reactors - Bardenpho System, therefore, this work intends an alternative system, where the anaerobic digestion and the denitrification happen in the same reactor reducing the number of reactors for two. The experimental system was constituted by two units: first one was a nitrification reactor with 35 L volume and 15 d of sludge age. This system was fed with raw sanitary waste. Second unit was an UASB, with 7.8 L and 6 h of hydraulic detention time, fed with ¾ of effluent nitrification reactor and ¼ of raw sanitary waste. This work had as objective to evaluate the performance of the UASB reactor. In terms of removal efficiency, of bath COD and nitrogen, it was verified that the anaerobic digestion process was not affected. The removal efficiency of organic material expressed in COD was 71%, performance already expected for a reactor of this type. It was also observed that the denitrification process happened; the removal nitrate efficiency was 90%. Therefore, the denitrification process in reactor UASB is viable.

Experimental and modelling study of drinking water hydrogenotrophic denitrification in packed-bed reactors

International Nuclear Information System (INIS)

Vasiliadou, I.A.; Karanasios, K.A.; Pavlou, S.; Vayenas, D.V.

2009-01-01

The aim of this work was to study hydrogenotrophic denitrification in packed-bed reactors under draw-fill and continuous operation. Three bench-scale packed-bed reactors with gravel in different sizes (mean diameter 1.75, 2.41 and 4.03 mm) as support media were used, in order to study the effect of particle size on reactors performance. The maximum denitrification rate achieved under draw-fill operation was 4.4 g NO 3 - -N/ld for the filter with gravel of 2.41 mm. This gravel size was chosen to perform experiments under continuous operation. Feed NO 3 - -N concentrations and hydraulic loadings (HL) ranged between 20-200 mg/l and 5.7-22.8 m 3 /m 2 d, respectively. A comparison between the two operating modes showed that, for low HL the draw-fill operation achieved higher denitrification rates, while for high HL and intermediate feed concentrations (40-60 mg NO 3 - -N/l) the continuous operation achieved higher denitrification rates (4.67-5.65 g/ld). Finally, experiments with three filters in series (with gravels of 4.03, 2.41 and 1.75 mm mean diameter) were also performed under continuous operation. The maximum denitrification rate achieved was 6.2 g NO 3 - -N/ld for feed concentration of 340 mg/l and HL of 11.5 m 3 /m 2 d. A model, which describes denitrification in packed-bed reactors, was also developed. The model predicts the concentration profiles of NO 3 - -N along filter height, in draw-fill as well as in continuous operation, satisfactorily.

Simultaneous desulfurization and denitrification by microwave reactor with ammonium bicarbonate and zeolite

International Nuclear Information System (INIS)

Wei Zaishan; Lin Zhehang; Niu Hejingying; He Haiming; Ji Yongfeng

2009-01-01

Microwave reactor with ammonium bicarbonate (NH 4 HCO 3 ) and zeolite was set up to study the simultaneous removal of sulfur dioxide (SO 2 ) and nitrogen oxides (NO x ) from flue gas. The results showed that the microwave reactor filled with NH 4 HCO 3 and zeolite could reduce SO 2 to sulfur with the best desulfurization efficiency of 99.1% and reduce NO x to nitrogen with the best NO x purifying efficiency of 86.5%. Microwave desulfurization and denitrification effect of the experiment using ammonium bicarbonate and zeolite together is much higher than that using ammonium bicarbonate or zeolite only. NO x concentration has little effect on denitrification but has no influence on desulfurization, SO 2 concentration has no effect on denitrification. The optimal microwave power and empty bed residence time (EBRT) on simultaneous desulfurization and dentrification are 211-280 W and 0.315 s, respectively. The mechanism for microwave reduced desulfurization and denitrification can be described as the microwave-induced catalytic reduction reaction between SO 2 , NO x and ammonium bicarbonate with zeolite being the catalyst and microwave absorbent

Isolation of a tyrosine-activating enzyme from baker's yeast

NARCIS (Netherlands)

Ven, A.M. van de; Koningsberger, V.V.; Overbeek, J.Th.G.

1958-01-01

The extracts of ether-CO2-frozen baker's yeast contain enzymes that catalyze the ATP-linked amino acid activation by way of pyrophosphate elimination. From the extract a tyrosine-activating enzyme could be isolated, which, judging from ultracentrifugation and electrophoretic data, was about 70% pure

Hydrogenotrophic denitrification in a packed bed reactor: effects of hydrogen-to-water flow rate ratio.

Science.gov (United States)

Lee, J W; Lee, K H; Park, K Y; Maeng, S K

2010-06-01

Hydrogen dissolution and hydrogenotrophic denitrification performance were investigated in a lab-scale packed bed reactor (PBR) by varying the hydrogen flow rate and hydraulic retention time (HRT). The denitrification performance was enhanced by increasing the hydrogen flow rate and HRT as a result of high dissolved hydrogen concentration (0.39mg/L) and utilization efficiencies (79%). In this study, the hydrogen-to-water flow rate ratio (Q(g)/Q(w)) was found to be a new operating factor representing the two parameters of hydrogen flow rate and HRT. Hydrogen dissolution and denitrification efficiency were nonlinearly and linearly correlated with the Q(g)/Q(w), respectively. Based on its excellent linear correlation with denitrification efficiency, Q(g)/Q(w) should be greater than 2.3 to meet the WHO's guideline of nitrate nitrogen for drinking water. This study demonstrates that Q(g)/Q(w) is a simple and robust factor to optimize hydrogen-sparged bioreactors for hydrogenotrophic denitrification. Copyright 2010 Elsevier Ltd. All rights reserved.

Nitrous oxide production in grassland soils: assessing the contribution of nitrifier denitrification

NARCIS (Netherlands)

Wrage, N.; Velthof, G.L.; Laanbroek, H.J.; Oenema, O.

2004-01-01

Nitrifier denitrification is the reduction of NO2- to N2 by nitrifiers. It leads to the production of the greenhouse gas nitrous oxide (N2O) as an intermediate and possible end product. It is not known how important nitrifier denitrification is for the production of N2O in soils. We explored N2O

Distribution of enzyme activity hotspots induced by earthworms in top- and subsoil

Science.gov (United States)

Hoang, D. T. T.

2016-12-01

Earthworms (Lumbricus terrestris L.) not only affect soil physics, but they also boost microbial activities and consequently create important hotspots of microbial mediated carbon and nutrient turnover through their burrowing activity. However, it is still unknown to which extend earthworms change the enzyme distribution and activity inside their burrows in top- and subsoil horizons. We hypothesized that earthworm burrows, which are enriched in available substrates, have higher percentage of enzyme activity hotspots than soil without earthworms, and that enzyme activities decreased with increasing depth because of the increasing recalcitrance of organic matter in subsoil. We visualized enzyme distribution inside and outside of worm burrows (biopores) by in situ soil zymography and measured enzyme kinetics of 6 enzymes - β-glucosidase (GLU), cellobiohydrolase (CBH), xylanase (XYL), chitinase (NAG), leucine aminopeptidase (LAP) and acid phosphatase (APT) - in pore and bulk soil material up to 105 cm. Zymography showed a heterogeneous distribution of hotspots in worm burrows. The hotspot areas was 2.4 to 14 times larger in the burrows than in soil without earthworms. However, the dispersion index of hotspot distribution showed more aggregated hotspots in soil without earthworms than in soil with earthworms and burrow wall. Enzyme activities decreased with depth, by a factor of 2 to 8 due to fresh C input from the soil surface. Compared to bulk soil, enzyme activities in topsoil biopores were up to 11 times higher for all enzymes, but in the subsoil activities of XYL, NAG and APT were lower in earthworm biopores than bulk soil. In conclusion, hotspots were twice as concentrated close to earthworm burrows as in surrounding soil. Earthworms exerted stronger effects on enzyme activities in biopores in the topsoil than in subsoil. Keywords: Earthworms, hotspots, enzyme activities, enzyme distribution, subsoil

Mixing-induced groundwater denitrification beneath a manured field in southern Alberta, Canada

International Nuclear Information System (INIS)

McCallum, J.E.; Ryan, M.C.; Mayer, B.; Rodvang, S.J.

2008-01-01

Contamination of shallow groundwater by NO 3 - from manure may occur under fields where manure is spread as fertilizer and for disposal. Attenuation of NO 3 - in groundwater occurs through denitrification under certain conditions, or NO 3 - -contaminated younger groundwater may mix with older groundwater, lowering the NO 3 - concentration. In this study, δ 15 N and δ 18 O values of NO 3 - , and δ 18 O and δ 2 H values in groundwater under a manured field were evaluated to determine if groundwater NO 3 - concentrations were influenced through mixing of shallower, manure-impacted groundwater with older groundwater, or if denitrification was reducing NO 3 - concentrations. The younger groundwater showed clear evidence of manure impact with elevated Cl - (∼85 mg L -1 ) and NO 3 - concentrations (∼50 mg NO 3 -N L -1 ), and δ 15 N and δ 18 O values of NO 3 - consistent with a manure source. Vertical hydraulic gradients and δ 18 O and δ 2 H values in groundwater suggest older, more reduced groundwater is upwelling locally and mixing with the shallow groundwater. Decreasing NO 3 :Cl ratios, decreasing dissolved O 2 concentrations, and increasing δ 15 N and δ 18 O values of NO 3 - suggest that denitrification occurs locally in the aquifer. The extent of denitrification is proportional to the fraction of deeper groundwater in the aquifer. Denitrification apparently does not proceed in the younger, manure-impacted groundwater in the absence of mixing

Chaperone-like activities of α-synuclein: α-Synuclein assists enzyme activities of esterases

International Nuclear Information System (INIS)

Ahn, Misun; Kim, SeungBum; Kang, Mira; Ryu, Yeonwoo; Doohun Kim, T.

2006-01-01

α-Synuclein, a major constituent of Lewy bodies (LBs), has been implicated to play a critical role in the pathogenesis of Parkinson's disease (PD), although the physiological function of α-synuclein has not yet been known. Here we have shown that α-synuclein, which has no well-defined secondary or tertiary structure, can protect the enzyme activity of microbial esterases against stress conditions such as heat, pH, and organic solvents. In particular, the flexibility of α-synuclein and its C-terminal region seems to be important for complex formation, but the structural integrity of the C-terminal region may not be required for stabilization of enzyme activity. In addition, atomic force microscopy (AFM) and in vivo enzyme assays showed highly specific interactions of esterases with α-synuclein. Our results indicate that α-synuclein not only protects the enzyme activity of microbial esterases in vitro, but also can stabilize the active conformation of microbial esterases in vivo

Endogenous influences on anammox and sulfocompound-oxidizing autotrophic denitrification coupling system (A/SAD) and dynamic operating strategy.

Science.gov (United States)

Sun, Xinbo; Du, Lingfeng; Hou, Yuqian; Cheng, Shaoju; Zhang, Xuxiang; Liu, Bo

2018-02-21

The anaerobic ammonia oxidation (anammox) and sulfocompound-oxidizing autotrophic denitrification coupling system (A/SAD) was initiated in an expanded granular sludge bed (EGSB) reactor for nitrogen removal from high-strength wastewater. Owing to cooperation between anammox and partial sulfocompound-oxidation autotrophic denitrification coupling system (PSAD), the highest nitrogen removal efficiency (NRE) of 98.1% ± 0.4% achieved at the optimal influent conditions of conversion efficiency of ammonium (CEA) of 55% and S 2 O 3 2- -S/NO 3 - -N (S/N) of 1.4 mol mol -1 . The activity of the short-cut sulfocompound-oxidizing autotrophic denitrification (SSAD) was also regulated to cope with dynamic CEA in the influent by changing the S/N, which was demonstrated to be effective in alleviating nitrite accumulation when the CEA was between 57% and 61%. Both the anammox and SAD bacteria enriched in the reactor after long-term incubation. Candidatus Brocadia and Candidatus Jettenia might be potentially contributing the most to anammox, while the Thiobacillus was the dominant taxa related to SAD. Copyright © 2018. Published by Elsevier Ltd.

Influence of 2. 45 GHz microwave radiation on enzyme activity

Energy Technology Data Exchange (ETDEWEB)

Galvin, M J; Parks, D L; McRee, D I

1981-05-01

The in vitro activity of acetylcholinesterase and creatine phosphokinase was determined during in vitro exposure to 2.45 GHz microwave radiation. The enzyme activities were examined during exposure to microwave radiation at specific absorption rates (SAR) of 1, 10, 50, and 100 mW/g. These specific absorption rates had no effect on the activity of either enzyme when the temperature of the control and exposed samples were similar. These data demonstrate that the activity of these two enzymes is not affected by microwave radiation at the SARs and frequency employed in this study.

HPRT Enzyme Activity of Blood Cells From Patients With Downs Syndrome

International Nuclear Information System (INIS)

Sbubber, E.K.; Abdul-Rahman, M.H.; Sultan, A.F.; Hamamy, H.A.

1998-01-01

Hypoxanthine phosphoribosyl transferase (HPRT) enzyme activity was determined in erythrocytes from 16 children (aged below one year to 11 year) with down s syndrome using 8-C 14 Hypoxanthine and radioeleelrophorsis techniques. Significant (P<0.01) reduction in HPRT enzyme activity was seen in D S children compared to that of 18 (age and sex matched) healthy children. Pure 21 - trisomic erythrocytes expressed lower enzyme activity than mosaic cell. Mothers of D S children showed significantly (P<0.01) lower enzyme activity than mothers of normal children . Reduced activity of HPRT enzyme was also observed in PHA-stimulated lymphocytes of DS children and their mothers. These results indicated that deficiency of HPRT in D S patients may contribute to the abnormal purine metabolism associated with the symptomatology of this syndrome

Exposure Time Distributions reveal Denitrification Rates along Groundwater Flow Path of an Agricultural Unconfined Aquifer

Science.gov (United States)

Kolbe, T.; Abbott, B. W.; Thomas, Z.; Labasque, T.; Aquilina, L.; Laverman, A.; Babey, T.; Marçais, J.; Fleckenstein, J. H.; Peiffer, S.; De Dreuzy, J. R.; Pinay, G.

2016-12-01

Groundwater contamination by nitrate is nearly ubiquitous in agricultural regions. Nitrate is highly mobile in groundwater and though it can be denitrified in the aquifer (reduced to inert N2 gas), this process requires the simultaneous occurrence of anoxia, an electron donor (e.g. organic carbon, pyrite), nitrate, and microorganisms capable of denitrification. In addition to this the ratio of the time groundwater spent in a denitrifying environment (exposure time) to the characteristic denitrification reaction time plays an important role, because denitrification can only occur if the exposure time is longer than the characteristic reaction time. Despite a long history of field studies and numerical models, it remains exceedingly difficult to measure or model exposure times in the subsurface at the catchment scale. To approach this problem, we developed a unified modelling approach combining measured environmental proxies with an exposure time based reactive transport model. We measured groundwater age, nitrogen and sulfur isotopes, and water chemistry from agricultural wells in an unconfined aquifer in Brittany, France, to quantify changes in nitrate concentration due to dilution and denitrification. Field data showed large differences in nitrate concentrations among wells, associated with differences in the exposure time distributions. By constraining a catchment-scale characteristic reaction time for denitrification with water chemistry proxies and exposure times, we were able to assess rates of denitrification along groundwater flow paths. This unified modeling approach is transferable to other catchments and could be further used to investigate how catchment structure and flow dynamics interact with biogeochemical processes such as denitrification.

Edaphic Conditions Regulate Denitrification Directly and Indirectly by Altering Denitrifier Abundance in Wetlands along the Han River, China.

Science.gov (United States)

Xiong, Ziqian; Guo, Laodong; Zhang, Quanfa; Liu, Guihua; Liu, Wenzhi

2017-05-16

Riparian wetlands play a critical role in retaining nitrogen (N) from upland runoff and improving river water quality, mainly through biological processes such as soil denitrification. However, the relative contribution of abiotic and biotic factors to riparian denitrification capacity remains elusive. Here we report the spatiotemporal dynamics of potential and unamended soil denitrification rates in 20 wetlands along the Han River, an important water source in central China. We also quantified the abundance of soil denitrifying microorganisms using nirK and nirS genes. Results showed that soil denitrification rates were significantly different between riparian and reservoir shoreline wetlands, but not between mountain and lowland wetlands. In addition, soil denitrification rates showed strong seasonality, with higher values in August (summer) and April (spring) but lower values in January (winter). The potential and unamended denitrification rates were positively correlated with edaphic conditions (moisture and carbon concentration), denitrifier abundance, and plant species richness. Path analysis further revealed that edaphic conditions could regulate denitrification rates both directly and indirectly through their effects on denitrifier abundance. Our findings highlight that not only environmental factors, but also biotic factors including denitrifying microorganisms and standing vegetation, play an important role in regulating denitrification rate and N removal capacity in riparian wetlands.

Chaperone-Like Activity of ß-Casein and Its Effect on Residual in Vitro Activity of Food Enzymes

DEFF Research Database (Denmark)

Sulewska, Anna Maria

ABSTRACT Activity of endogenous enzymes may cause browning of fruits and vegetables. These enzymes can be inactivated, for example by heat treatment, but the response of enzymes to heat treatment depends on many factors. Foods are very complex systems and the stability of enzymes......-casein on the enzymatic activity of three targets was tested by monitoring enzyme activity after heat treatment and by measuring the intensity of scattered light during and after heat treatment. β-Casein was shown to interact at elevated temperatures with three selected targets:horseradish peroxidase, tyrosinase from......, residual activity of horseradish peroxidase was lower in samples containing BSA than in samples without any addition. Horseradish peroxidase heated with BSA did not regain activity within one hour after treatment. BSA is often added to enzyme solutions to prevent enzyme adhesion to vial surfaces...

Regulation by sliding operation of a denitrification process; Regulation par regime glissant d`un procede de denitrification

Energy Technology Data Exchange (ETDEWEB)

Babary, J.P.; Bourrel, S.

1995-12-31

Biotechnological processing of waste waters is made through fixed bed bioreactors. Biochemical reaction kinetics can be modelled by non linear differential equations. The model is used to develop control laws. As an application, a denitrification bioreactor is modelled. Then control algorithm is checked through mathematical simulation of nitrogen compounds concentration ratio. (D.L.)

Concentration profiles near an activated enzyme.

Science.gov (United States)

Park, Soohyung; Agmon, Noam

2008-09-25

When a resting enzyme is activated, substrate concentration profile evolves in its vicinity, ultimately tending to steady state. We use modern theories for many-body effects on diffusion-influenced reactions to derive approximate analytical expressions for the steady-state profile and the Laplace transform of the transient concentration profiles. These show excellent agreement with accurate many-particle Brownian-dynamics simulations for the Michaelis-Menten kinetics. The steady-state profile has a hyperbolic dependence on the distance of the substrate from the enzyme, albeit with a prefactor containing the complexity of the many-body effects. These are most conspicuous for the substrate concentration at the surface of the enzyme. It shows an interesting transition as a function of the enzyme turnover rate. When it is high, the contact concentration decays monotonically to steady state. However, for slow turnover it is nonmonotonic, showing a minimum due to reversible substrate binding, then a maximum due to diffusion of new substrate toward the enzyme, and finally decay to steady state. Under certain conditions one can obtain a good estimate for the critical value of the turnover rate constant at the transition.

Extraction of Active Enzymes from "Hard-to-Break-Cells"

DEFF Research Database (Denmark)

Ottaviani, Alessio; Tesauro, Cinzia; Fjelstrup, S

We present the utilization of a rolling circle amplification (RCA) based assay to investigate the extraction efficiency of active enzymes from a class of “hard-to-break” cells, yeast Saccaramyces cerevisiae. Current analyses of microorganisms, such as pathogenic bacteria, parasites or particular...... life stages of microorganisms (e.g. spores from bacteria or fungi) is hampered by the lack of efficient lysis protocols that preserve the activity and integrity of the cellular content. Presented herein is a flexible scheme to screen lysis protocols for active enzyme extraction. We also report a gentle...... yet effective approach for extraction of active enzymes by entrapping cells in microdroplets. Combined effort of optimized extraction protocols and effective analytical approaches is expected to generate impact in future disease diagnosis and environmental safety....

Stimulation of Escherichia coli DNA photoreactivating enzyme activity by adenosine 5'-triphosphate

International Nuclear Information System (INIS)

Koka, P.

1984-01-01

A purification procedure consisting of Biorex-70, single-stranded DNA-agarose, and ultraviolet (UV) light irradiated DNA-cellulose chromatography has been adopted for the Escherichia coli photoreactivating enzyme, to obtain enzyme preparations that are free of extraneous nucleic acid or nucleotides. The purification yields high specific activities (75 000 pmol h -1 mg -1 ) with a 50% recovery. Enzyme preparations have also been obtained from UV-irradiated DNA-cellulose by exposure to visible light. These enzyme preparations contain oligoribonucleotides, up to 26 nucleotides in length in relation to DNA size markers, but these are not essential for enzymatic activity. When the enzyme is preincubated with exogenous ATP a 10-fold stimulation in the enzyme activity has been observed. It has been determined by polyacrylamide gel electrophoresis and high-voltage diethylaminoethyl paper electrophoresis that the light-released enzyme samples from a preincubated and washed mixture of the enzyme, [γ- 32 P]ATP, and UV-irradiated DNA-cellulose contained exogenous [γ- 32 P], which eluted with the enzyme-containing fractions when subjected to Bio-Gel P-30 chromatography. GTP caused a slight enhancement of the enzyme activity while ADP strongly inhibited photoreactivation, at the same concentration and conditions. Higher (X5) concentrations of ADP and adenosine 5'-(β, γ-methylenetriphosphate) totally inhibited the enzyme activity. Dialysis of a photoreactivating enzyme preparation against a buffer solution containing 1 mM ATP caused a 9-fold stimulation of the enzyme activity. In addition, there is an apparent hydrolysis of ATP during photoreactivation as measured by the release of 32 P from [γ- 32 P]ATP

Effects of aquatic vegetation type on denitrification

NARCIS (Netherlands)

Veraart, A.J.; Bruijne, de W.J.J.; Peeters, E.T.H.M.; Klein, de J.J.M.; Scheffer, M.

2011-01-01

In a microcosm 15N enrichment experiment we tested the effect of floating vegetation (Lemna sp.) and submerged vegetation (Elodea nuttallii) on denitrification rates, and compared it to systems without macrophytes. Oxygen concentration, and thus photosynthesis, plays an important role in regulating

Enzyme activity measurement via spectral evolution profiling and PARAFAC

DEFF Research Database (Denmark)

Baum, Andreas; Meyer, Anne S.; Garcia, Javier Lopez

2013-01-01

The recent advances in multi-way analysis provide new solutions to traditional enzyme activity assessment. In the present study enzyme activity has been determined by monitoring spectral changes of substrates and products in real time. The method relies on measurement of distinct spectral...... fingerprints of the reaction mixture at specific time points during the course of the whole enzyme catalyzed reaction and employs multi-way analysis to detect the spectral changes. The methodology is demonstrated by spectral evolution profiling of Fourier Transform Infrared (FTIR) spectral fingerprints using...

Patterns of functional enzyme activity in fungus farming ambrosia beetles.

Science.gov (United States)

De Fine Licht, Henrik H; Biedermann, Peter H W

2012-06-06

In wood-dwelling fungus-farming weevils, the so-called ambrosia beetles (Curculionidae: Scolytinae and Platypodinae), wood in the excavated tunnels is used as a medium for cultivating fungi by the combined action of digging larvae (which create more space for the fungi to grow) and of adults sowing and pruning the fungus. The beetles are obligately dependent on the fungus that provides essential vitamins, amino acids and sterols. However, to what extent microbial enzymes support fungus farming in ambrosia beetles is unknown. Here we measure (i) 13 plant cell-wall degrading enzymes in the fungus garden microbial consortium of the ambrosia beetle Xyleborinus saxesenii, including its primary fungal symbionts, in three compartments of laboratory maintained nests, at different time points after gallery foundation and (ii) four specific enzymes that may be either insect or microbially derived in X. saxesenii adult and larval individuals. We discovered that the activity of cellulases in ambrosia fungus gardens is relatively small compared to the activities of other cellulolytic enzymes. Enzyme activity in all compartments of the garden was mainly directed towards hemicellulose carbohydrates such as xylan, glucomannan and callose. Hemicellulolytic enzyme activity within the brood chamber increased with gallery age, whereas irrespective of the age of the gallery, the highest overall enzyme activity were detected in the gallery dump material expelled by the beetles. Interestingly endo-β-1,3(4)-glucanase activity capable of callose degradation was identified in whole-body extracts of both larvae and adult X. saxesenii, whereas endo-β-1,4-xylanase activity was exclusively detected in larvae. Similar to closely related fungi associated with bark beetles in phloem, the microbial symbionts of ambrosia beetles hardly degrade cellulose. Instead, their enzyme activity is directed mainly towards comparatively more easily accessible hemicellulose components of the ray

A Simple and Accurate Method for Measuring Enzyme Activity.

Science.gov (United States)

Yip, Din-Yan

1997-01-01

Presents methods commonly used for investigating enzyme activity using catalase and presents a new method for measuring catalase activity that is more reliable and accurate. Provides results that are readily reproduced and quantified. Can also be used for investigations of enzyme properties such as the effects of temperature, pH, inhibitors,…

The use of a rubble chimney for denitrification of irrigation return waters

Energy Technology Data Exchange (ETDEWEB)

Evans, Roy B; Kruger, Paul [Civil Engineering Department, Stanford University (United States)

1970-05-15

Biological denitrification has been proposed as a means of removing nitrates from waste waters to control eutrophication in receiving waters. A potential use for this method is the treatment of irrigation return waters containing high concentrations of nitrate-nitrogen, since direct discharge of such wastes may cause objectionable algal growth in the receiving waters. For example, the process may be used to treat agricultural waste waters in the San Joaquin Valley in California, where an estimated 580,000 acre-feet/year of return waters, containing 20 mg/l of nitrate-nitrogen, will require disposal by A.D. 2020. Two methods of biological denitrification are presently under study for possible use in the San Joaquin Valley. In one method nitrates are reduced to nitrogen gas by bacterial action in deep ponds; in the other method bacterial denitrification takes place in biological filters. In biological filters, bacteria are grown on columns of submerged stones. A possible alternative to the conventional construction of these filters is the creation of a rubble chimney by a contained nuclear explosion. This paper presents the results of a preliminary investigation of the feasibility of using a rubble chimney as a biological filter for denitrification. (author)

Simultaneous desulfurization and denitrification by microwave reactor with ammonium bicarbonate and zeolite

Energy Technology Data Exchange (ETDEWEB)

Wei Zaishan [School of Environmental Science and Engineering, Sun Yat-sen University, Guangzhou 510275 (China)], E-mail: weizaishan98@163.com; Lin Zhehang; Niu Hejingying; He Haiming; Ji Yongfeng [School of Environmental Science and Engineering, Sun Yat-sen University, Guangzhou 510275 (China)

2009-03-15

Microwave reactor with ammonium bicarbonate (NH{sub 4}HCO{sub 3}) and zeolite was set up to study the simultaneous removal of sulfur dioxide (SO{sub 2}) and nitrogen oxides (NO{sub x}) from flue gas. The results showed that the microwave reactor filled with NH{sub 4}HCO{sub 3} and zeolite could reduce SO{sub 2} to sulfur with the best desulfurization efficiency of 99.1% and reduce NO{sub x} to nitrogen with the best NO{sub x} purifying efficiency of 86.5%. Microwave desulfurization and denitrification effect of the experiment using ammonium bicarbonate and zeolite together is much higher than that using ammonium bicarbonate or zeolite only. NO{sub x} concentration has little effect on denitrification but has no influence on desulfurization, SO{sub 2} concentration has no effect on denitrification. The optimal microwave power and empty bed residence time (EBRT) on simultaneous desulfurization and dentrification are 211-280 W and 0.315 s, respectively. The mechanism for microwave reduced desulfurization and denitrification can be described as the microwave-induced catalytic reduction reaction between SO{sub 2}, NO{sub x} and ammonium bicarbonate with zeolite being the catalyst and microwave absorbent.

The use of a rubble chimney for denitrification of irrigation return waters

International Nuclear Information System (INIS)

Evans, Roy B.; Kruger, Paul

1970-01-01

Biological denitrification has been proposed as a means of removing nitrates from waste waters to control eutrophication in receiving waters. A potential use for this method is the treatment of irrigation return waters containing high concentrations of nitrate-nitrogen, since direct discharge of such wastes may cause objectionable algal growth in the receiving waters. For example, the process may be used to treat agricultural waste waters in the San Joaquin Valley in California, where an estimated 580,000 acre-feet/year of return waters, containing 20 mg/l of nitrate-nitrogen, will require disposal by A.D. 2020. Two methods of biological denitrification are presently under study for possible use in the San Joaquin Valley. In one method nitrates are reduced to nitrogen gas by bacterial action in deep ponds; in the other method bacterial denitrification takes place in biological filters. In biological filters, bacteria are grown on columns of submerged stones. A possible alternative to the conventional construction of these filters is the creation of a rubble chimney by a contained nuclear explosion. This paper presents the results of a preliminary investigation of the feasibility of using a rubble chimney as a biological filter for denitrification. (author)

Evolutionary transitions in enzyme activity of ant fungus gardens.

Science.gov (United States)

De Fine Licht, Henrik H; Schiøtt, Morten; Mueller, Ulrich G; Boomsma, Jacobus J

2010-07-01

Fungus-growing (attine) ants and their fungal symbionts passed through several evolutionary transitions during their 50 million year old evolutionary history. The basal attine lineages often shifted between two main cultivar clades, whereas the derived higher-attine lineages maintained an association with a monophyletic clade of specialized symbionts. In conjunction with the transition to specialized symbionts, the ants advanced in colony size and social complexity. Here we provide a comparative study of the functional specialization in extracellular enzyme activities in fungus gardens across the attine phylogeny. We show that, relative to sister clades, gardens of higher-attine ants have enhanced activity of protein-digesting enzymes, whereas gardens of leaf-cutting ants also have increased activity of starch-digesting enzymes. However, the enzyme activities of lower-attine fungus gardens are targeted primarily toward partial degradation of plant cell walls, reflecting a plesiomorphic state of nondomesticated fungi. The enzyme profiles of the higher-attine and leaf-cutting gardens appear particularly suited to digest fresh plant materials and to access nutrients from live cells without major breakdown of cell walls. The adaptive significance of the lower-attine symbiont shifts remains unclear. One of these shifts was obligate, but digestive advantages remained ambiguous, whereas the other remained facultative despite providing greater digestive efficiency.

Simultaneous denitrification and anaerobic digestion in GRAnular Bed Baffled Reactor (GRABBR)

International Nuclear Information System (INIS)

Baloch, M.I.; Akunna, J.C.

2002-01-01

This study elucidates the characteristics of compartmentalised anaerobic system seeded with UASB granules, called GRAanular Bed Baffled Reactor (GRABBR), for combined denitrification and anaerobic digestion processes. The reactor was used for the treatment of glucose enriched synthetic wastewater with various nitrate concentrations. The study was carried out with a 10 litre working volume GRABBR divided into 5 equal compartments operating at organic loading rate (OLR) of 20 kg COD/m 3 .d with a hydraulic retention time (HRT) of 6 hours. At these conditions, phase separation (between acidogenesis and methanogenesis) was created in the system and then the effect of varying nitrate concentrations (50-200 mg/l NO 3 -N) in the acidogenic zone (i.e. first compartment) was studied. Due to its unique compartmentalised design, denitrification was the major pathway for nitrate reduction with no noticeable dissimilatory nitrate reduction to ammonia (DNRA). More than 84% of all added nitrates were removed in the acidogenic zone, showing that acidogens possess high denitrifying capabilities. The denitrification rate increased with increase in nitrate concentration, with maximum value estimated as 175 mg NO 3 -N/l.h at influent nitrate concentration of 200 mg/l NO 3 -N in the acidogenic zone. Although nitrate addition resulted in lower methane production, COD removal efficiencies improved by up to 8% when compared with the reactor performance before nitrate addition. Furthermore, the alkalinity produced during denitrification improved the stability of the system by controlling the decrease in pH resulting from acidogenesis. The system encouraged simultaneous denitrification and anaerobic digestion in a single unit by accommodating denitrifiers in the early compartments and allowing methanogenesis to flourish in the downstream compartments of the system, thus minimising inhibition to methane producing bacteria by nitrates. (author)

Effects of specific inhibitors on anammox and denitrification in marine sediments.

Science.gov (United States)

Jensen, Marlene Mark; Thamdrup, Bo; Dalsgaard, Tage

2007-05-01

The effects of three metabolic inhibitors (acetylene, methanol, and allylthiourea [ATU]) on the pathways of N2 production were investigated by using short anoxic incubations of marine sediment with a 15N isotope technique. Acetylene inhibited ammonium oxidation through the anammox pathway as the oxidation rate decreased exponentially with increasing acetylene concentration; the rate decay constant was 0.10+/-0.02 microM-1, and there was 95% inhibition at approximately 30 microM. Nitrous oxide reduction, the final step of denitrification, was not sensitive to acetylene concentrations below 10 microM. However, nitrous oxide reduction was inhibited by higher concentrations, and the sensitivity was approximately one-half the sensitivity of anammox (decay constant, 0.049+/-0.004 microM-1; 95% inhibition at approximately 70 microM). Methanol specifically inhibited anammox with a decay constant of 0.79+/-0.12 mM-1, and thus 3 to 4 mM methanol was required for nearly complete inhibition. This level of methanol stimulated denitrification by approximately 50%. ATU did not have marked effects on the rates of anammox and denitrification. The profile of inhibitor effects on anammox agreed with the results of studies of the process in wastewater bioreactors, which confirmed the similarity between the anammox bacteria in bioreactors and natural environments. Acetylene and methanol can be used to separate anammox and denitrification, but the effects of these compounds on nitrification limits their use in studies of these processes in systems where nitrification is an important source of nitrate. The observed differential effects of acetylene and methanol on anammox and denitrification support our current understanding of the two main pathways of N2 production in marine sediments and the use of 15N isotope methods for their quantification.

Integrating anammox with the autotrophic denitrification process via electrochemistry technology.

Science.gov (United States)

Qiao, Sen; Yin, Xin; Zhou, Jiti; Wei, Li'e; Zhong, Jiayou

2018-03-01

In this study, an autotrophic denitrification process was successfully coupled with anammox to remove the nitrate by-product via electrochemical technology. When the voltage applied to the combined electrode reactor was 1.5 V, the electrode reaction removed nitrate by using the autotrophic denitrification biomass without affecting the anammox biomass. The nitrogen removal efficiency of the combined electrode reactor reached 99.1% without detectable nitrate at an influent NO 2 - -N/NH 4 + -N ratio of 1.5. On day 223, using the model calculations based on reaction equations, 19.7% of total nitrogen was removed via the autotrophic denitrification process, while the majority of nitrogen removal (approximately 79.4%) was attributed to the anammox reaction. Small variations of the population numbers and community structure of artificial bacteria according to electron microscopy predicted that the anammox and autotrophic denitrifying biomasses could coexist in the electrode reactor. Then, 16S rRNA analysis determined that the anammox biomass group was always dominant in mixed flora during continuous cultivation. Copyright © 2017 Elsevier Ltd. All rights reserved.

Light-regulation of enzyme activity in anacystis nidulans (Richt.).

Science.gov (United States)

Duggan, J X; Anderson, L E

1975-01-01

The effect of light on the levels of activity of six enzymes which are light-modulated in higher plants was examined in the photosynthetic procaryot Anacystis nidulans. Ribulose-5-phosphate kinase (EC 2.7.1.19) was found to be light-activated in vivo and dithiothreitol-activated in vitro while glucose-6-phosphate dehydrogenase (EC 1.1.1.49) was light-inactivated and dithiothreitol-inactivated. The enzymes fructose-1,6-diphosphate phosphatase (EC 3.1.3.11), sedoheptulose-1,7-diphosphate phosphatase, NAD- and NADP-linked glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.12; EC 1.2.1.13) were not affected by light treatment of the intact algae, but sedoheptulose-diphosphate phosphatase and the glyceraldehyde-3-phosphate dehydrogenases were dithiothreitol-activated in crude extracts. Light apparently controls the activity of the reductive and oxidative pentose phosphate pathway in this photosynthetic procaryot as in higher plants, through a process which probably involves reductive modulation of enzyme activity.

Activity enhancement of ligninolytic enzymes of Trametes versicolor ...

African Journals Online (AJOL)

Suspended cultures of white-rot fungus, Trametes versicolor, supplemented with bagasse powder showed a concentration dependent enhancement in the ligninolytic enzymes activity in liquid shake cultures. 2% (w/v) bagasse powder improved greater stability to the enzymes. The optimum pH is 3.5 and the optimum ...

Temperature and UV light affect the activity of marine cell-free enzymes

Directory of Open Access Journals (Sweden)

B. Thomson

2017-09-01

Full Text Available Microbial extracellular enzymatic activity (EEA is the rate-limiting step in the degradation of organic matter in the oceans. These extracellular enzymes exist in two forms: cell-bound, which are attached to the microbial cell wall, and cell-free, which are completely free of the cell. Contrary to previous understanding, cell-free extracellular enzymes make up a substantial proportion of the total marine EEA. Little is known about these abundant cell-free enzymes, including what factors control their activity once they are away from their sites (cells. Experiments were run to assess how cell-free enzymes (excluding microbes respond to ultraviolet radiation (UVR and temperature manipulations, previously suggested as potential control factors for these enzymes. The experiments were done with New Zealand coastal waters and the enzymes studied were alkaline phosphatase (APase, β-glucosidase, (BGase, and leucine aminopeptidase (LAPase. Environmentally relevant UVR (i.e. in situ UVR levels measured at our site reduced cell-free enzyme activities by up to 87 % when compared to controls, likely a consequence of photodegradation. This effect of UVR on cell-free enzymes differed depending on the UVR fraction. Ambient levels of UV radiation were shown to reduce the activity of cell-free enzymes for the first time. Elevated temperatures (15 °C increased the activity of cell-free enzymes by up to 53 % when compared to controls (10 °C, likely by enhancing the catalytic activity of the enzymes. Our results suggest the importance of both UVR and temperature as control mechanisms for cell-free enzymes. Given the projected warming ocean environment and the variable UVR light regime, it is possible that there could be major changes in the cell-free EEA and in the enzymes contribution to organic matter remineralization in the future.

A Review on the Effects of Supercritical Carbon Dioxide on Enzyme Activity

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Marie Zarevúcka

2010-01-01

Full Text Available Different types of enzymes such as lipases, several phosphatases, dehydrogenases, oxidases, amylases and others are well suited for the reactions in SC-CO2. The stability and the activity of enzymes exposed to carbon dioxide under high pressure depend on enzyme species, water content in the solution and on the pressure and temperature of the reaction system. The three-dimensional structure of enzymes may be significantly altered under extreme conditions, causing their denaturation and consequent loss of activity. If the conditions are less adverse, the protein structure may be largely retained. Minor structural changes may induce an alternative active protein state with altered enzyme activity, specificity and stability.

Excessive use of nitrogen in Chinese agriculture results in high N2O/(N2O+N2) product ratio of denitrification, primarily due to acidification of the soils

Science.gov (United States)

Qu, Zhi; Wang, Jingguo; Almøy, Trygve; Bakken, Lars R

2014-01-01

China is the world's largest producer and consumer of fertilizer N, and decades of overuse has caused nitrate leaching and possibly soil acidification. We hypothesized that this would enhance the soils' propensity to emit N2O from denitrification by reducing the expression of the enzyme N2O reductase. We investigated this by standardized oxic/anoxic incubations of soils from five long-term fertilization experiments in different regions of China. After adjusting the nitrate concentration to 2 mM, we measured oxic respiration (R), potential denitrification (D), substrate-induced denitrification, and the denitrification product stoichiometry (NO, N2O, N2). Soils with a history of high fertilizer N levels had high N2O/(N2O+N2) ratios, but only in those field experiments where soil pH had been lowered by N fertilization. By comparing all soils, we found a strong negative correlation between pH and the N2O/(N2O+N2) product ratio (r2 = 0.759, P soil pH. The immediate effect of liming acidified soils was lowered N2O/(N2O+N2) ratios. The results provide evidence that soil pH has a marginal direct effect on potential denitrification, but that it is the master variable controlling the percentage of denitrified N emitted as N2O. It has been known for long that low pH may result in high N2O/(N2O+N2) product ratios of denitrification, but our documentation of a pervasive pH-control of this ratio across soil types and management practices is new. The results are in good agreement with new understanding of how pH may interfere with the expression of N2O reductase. We argue that the management of soil pH should be high on the agenda for mitigating N2O emissions in the future, particularly for countries where ongoing intensification of plant production is likely to acidify the soils. PMID:24249526

Patterns of functional enzyme activity in fungus farming ambrosia beetles

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De Fine Licht Henrik H

2012-06-01

Full Text Available Abstract Introduction In wood-dwelling fungus-farming weevils, the so-called ambrosia beetles (Curculionidae: Scolytinae and Platypodinae, wood in the excavated tunnels is used as a medium for cultivating fungi by the combined action of digging larvae (which create more space for the fungi to grow and of adults sowing and pruning the fungus. The beetles are obligately dependent on the fungus that provides essential vitamins, amino acids and sterols. However, to what extent microbial enzymes support fungus farming in ambrosia beetles is unknown. Here we measure (i 13 plant cell-wall degrading enzymes in the fungus garden microbial consortium of the ambrosia beetle Xyleborinus saxesenii, including its primary fungal symbionts, in three compartments of laboratory maintained nests, at different time points after gallery foundation and (ii four specific enzymes that may be either insect or microbially derived in X. saxesenii adult and larval individuals. Results We discovered that the activity of cellulases in ambrosia fungus gardens is relatively small compared to the activities of other cellulolytic enzymes. Enzyme activity in all compartments of the garden was mainly directed towards hemicellulose carbohydrates such as xylan, glucomannan and callose. Hemicellulolytic enzyme activity within the brood chamber increased with gallery age, whereas irrespective of the age of the gallery, the highest overall enzyme activity were detected in the gallery dump material expelled by the beetles. Interestingly endo-β-1,3(4-glucanase activity capable of callose degradation was identified in whole-body extracts of both larvae and adult X. saxesenii, whereas endo-β-1,4-xylanase activity was exclusively detected in larvae. Conclusion Similar to closely related fungi associated with bark beetles in phloem, the microbial symbionts of ambrosia beetles hardly degrade cellulose. Instead, their enzyme activity is directed mainly towards comparatively more easily

Denitrification, anammox and fixed nitrogen removal in the water column of a tropical great lake

Science.gov (United States)

Darchambeau, François; Roland, Fleur; Crowe, Sean A.; De Brabandere, Loreto; Llirós, Marc; Garcia-Armisen, Tamara; Inceoglu, Ozgul; Michiels, Céline; Servais, Pierre; Morana, Cédric D. T.; Bouillon, Steven; Meysman, Filip; Veuger, Bart; Masilya, Pascal M.; Descy, Jean-Pierre; Borges, Alberto V.

2013-04-01

If rates of microbial denitrification in aquatic systems are poorly constrained, it is much more the case for tropical water bodies. Lake Kivu [2.50° S 1.59° S, 29.37° E 28.83° E] is one of the great lakes of the East African Rift. It is an oligotrophic lake characterized by anoxic deep waters rich in dissolved gases (methane and carbon dioxide) and nutrients, and by well oxygenated and nutrient-depleted surface waters. During the seasonally stratified rainy season (October to May), a nitrogenous zone characterized by the accumulation of nitrite (NO2-) and nitrate (NO3-) is often observed in the lower layer of the mixolimnion. It results from nitrification of ammonium released by decaying organic matter. With the seasonal uplift of the oxygen minimum zone, the nitrogenous zone becomes anoxic and might be the most preferential area for fixed nitrogen (N) removal in Lake Kivu. Our work aimed at identifying and quantifying the processes of N losses by denitrification and/or anammox in the nitrogenous zone of the Lake Kivu water column. During 5 sampling campaigns (March 2010, October 2010, June 2011, February 2012 and September 2012), isotopic labelling experiments were used to quantify denitrification and anammox rates along vertical profiles at two pelagic stations of the main lake. Moreover, N2:Ar ratios were estimated during the September 2012 campaign, and 16S rDNA pyrosequencing was used to describe bacterial community composition during the last 2 campaigns. No bacteria related to organisms performing anammox was observed and labelling experiments failed to detect anammox at any locations and any depths. In Lake Kivu, denitrifying bacteria were mainly related to Denitratisoma and Thiobacillus genus. Significant denitrification rates were observed at several occasions, especially under the oxic-anoxic interface in the bottom of the nitracline. The annual average denitrification rate was estimated at ~150 μmoles N m-2 d-1. Denitrification was not the only

Enzyme Stability and Activity in Non-Aqueous Reaction Systems: A Mini Review

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Shihui Wang

2016-02-01

Full Text Available Enormous interest in biocatalysis in non-aqueous phase has recently been triggered due to the merits of good enantioselectivity, reverse thermodynamic equilibrium, and no water-dependent side reactions. It has been demonstrated that enzyme has high activity and stability in non-aqueous media, and the variation of enzyme activity is attributed to its conformational modifications. This review comprehensively addresses the stability and activity of the intact enzymes in various non-aqueous systems, such as organic solvents, ionic liquids, sub-/super-critical fluids and their combined mixtures. It has been revealed that critical factors such as Log P, functional groups and the molecular structures of the solvents define the microenvironment surrounding the enzyme molecule and affect enzyme tertiary and secondary structure, influencing enzyme catalytic properties. Therefore, it is of high importance for biocatalysis in non-aqueous media to elucidate the links between the microenvironment surrounding enzyme surface and its stability and activity. In fact, a better understanding of the correlation between different non-aqueous environments and enzyme structure, stability and activity can contribute to identifying the most suitable reaction medium for a given biotransformation.

Visualization of enzyme activities inside earthworm biopores by in situ soil zymography

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Thu Duyen Hoang, Thi; Razavi, Bahar. S.; Blagodatskaya, Evgenia; Kuzyakov, Yakov

2015-04-01

Earthworms can strongly activate microorganisms, increase microbial and enzyme activities and consequently the turnover of native soil organic matter. In extremely dynamic microhabitats and hotspots as biopores made by earthworms, the in situ enzyme activities are a footprint of complex biotic interactions. The effect of earthworms on the alteration of enzyme activities inside biopores and the difference between bio-pores and earthworm-free soil was visualized by in situ soil zymography (Spohn and Kuzyakov, 2014). For the first time, we prepared quantitative imaging of enzyme activities in biopores. Furthermore, we developed the zymography technique by direct application of a substrate saturated membrane to the soil to obtain better spatial resolution. Lumbricus terrestris L. was placed into transparent box (15×20×15cm). Simultaneously, maize seed was sown in the soil. Control soil box with maize and without earthworm was prepared in the same way. After two weeks when bio-pore systems were formed by earthworm, we visualized in situ enzyme activities of five hydrolytic enzymes (β-glucosidase, cellobiohydrolase, chitinase, xylanase, leucine aminopeptidase) and phosphatase. Followed by non-destructive zymography, biopore samples and control soil were destructively collected to assay enzyme kinetics by fluorogenically labeled substrates method. Zymography showed higher activity of β-glucosidase, chitinase, xylanase and phosphatase in biopores comparing to bulk soil. These differences were further confirmed by fluorimetric microplate enzyme assay detected significant difference of Vmax in four above mentioned enzymes. Vmax of β-glucosidase, chitinase, xylanase and phosphatase in biopores is 68%, 108%, 50% and 49% higher than that of control soil. However, no difference in cellobiohydrolase and leucine aminopeptidase kinetics between biopores and control soil were detected. This indicated little effect of earthworms on protein and cellulose transformation in soil

Effect of pine bark and compost on the biological denitrification process of non-hazardous landfill leachate: Focus on the microbiology

International Nuclear Information System (INIS)

Trois, Cristina; Coulon, Frederic; Polge de Combret, Cecile; Martins, Jean M.F.; Oxarango, Laurent

2010-01-01

In an attempt to optimize the cost-efficiency of landfill leachate treatment by biological denitrification process, our study focused on finding low-cost alternatives to traditional expensive chemicals such as composted garden refuse and pine bark, which are both available in large amount in South African landfill sites. The overall objective was to assess the behaviour of the bacterial community in relation to each substrate while treating high strength landfill leachates. Denitrification processes in fixed bed reactors were simulated at laboratory scale using anaerobic batch tests with immature compost and pine bark. High strength leachate was simulated using a solution of water and nitrate at a concentration of 500 mg l -1 . Results suggest that pine bark released large amounts of phenolic compounds and hydroxylated benzene rings, which both can delay the acclimatization time and inhibit the biological denitrification (only 30% efficiency). Furthermore, presence of potential pathogens like Enterobacter and Pantoea agglomerans prevents the applicability of the pine bark in full-scale operations. On the other hand, lightly composted garden refuse (CGR) offered an adequate substrate for the formation of a biofilm necessary to complete the denitrification process (total nitrate removal observed within 7 days). CGR further contributed to a rapid establishment of an active consortium of denitrifiers including Acinetobacter, Rhizobium, Thermomonas, Rheinheimera, Phaeospirillum and Flavobacterium. Clearly the original composition, nature, carbon to nitrogen ratio (C/N) and degree of maturity and stability of the substrates play a key role in the denitrification process, impacting directly on the development of the bacterial population and, therefore, on the long-term removal efficiency.

Effect of pine bark and compost on the biological denitrification process of non-hazardous landfill leachate: Focus on the microbiology

Energy Technology Data Exchange (ETDEWEB)

Trois, Cristina, E-mail: troisc@ukzn.ac.za [Centre for Research in Environmental, Coastal and Hydrological Engineering, School of Civil Engineering, Surveying and Construction, University of KwaZulu-Natal, Howard College Campus, Durban 4041 (South Africa); Coulon, Frederic; Polge de Combret, Cecile [Centre for Resource Management and Efficiency, School of Applied Sciences, Cranfield University, MK43 0AL (United Kingdom); Martins, Jean M.F.; Oxarango, Laurent [Laboratoire d' etude de Transferts en Hydrologie et Environnement, UMR 5564 (CNRS/INPG/IRD/UJF), Universite de Grenoble, BP 53, 38041 Grenoble Cedex 9 (France)

2010-09-15

In an attempt to optimize the cost-efficiency of landfill leachate treatment by biological denitrification process, our study focused on finding low-cost alternatives to traditional expensive chemicals such as composted garden refuse and pine bark, which are both available in large amount in South African landfill sites. The overall objective was to assess the behaviour of the bacterial community in relation to each substrate while treating high strength landfill leachates. Denitrification processes in fixed bed reactors were simulated at laboratory scale using anaerobic batch tests with immature compost and pine bark. High strength leachate was simulated using a solution of water and nitrate at a concentration of 500 mg l{sup -1}. Results suggest that pine bark released large amounts of phenolic compounds and hydroxylated benzene rings, which both can delay the acclimatization time and inhibit the biological denitrification (only 30% efficiency). Furthermore, presence of potential pathogens like Enterobacter and Pantoea agglomerans prevents the applicability of the pine bark in full-scale operations. On the other hand, lightly composted garden refuse (CGR) offered an adequate substrate for the formation of a biofilm necessary to complete the denitrification process (total nitrate removal observed within 7 days). CGR further contributed to a rapid establishment of an active consortium of denitrifiers including Acinetobacter, Rhizobium, Thermomonas, Rheinheimera, Phaeospirillum and Flavobacterium. Clearly the original composition, nature, carbon to nitrogen ratio (C/N) and degree of maturity and stability of the substrates play a key role in the denitrification process, impacting directly on the development of the bacterial population and, therefore, on the long-term removal efficiency.

European-scale modelling of groundwater denitrification and associated N2O production

International Nuclear Information System (INIS)

Keuskamp, J.A.; Drecht, G. van; Bouwman, A.F.

2012-01-01

This paper presents a spatially explicit model for simulating the fate of nitrogen (N) in soil and groundwater and nitrous oxide (N 2 O) production in groundwater with a 1 km resolution at the European scale. The results show large heterogeneity of nitrate outflow from groundwater to surface water and production of N 2 O. This heterogeneity is the result of variability in agricultural and hydrological systems. Large parts of Europe have no groundwater aquifers and short travel times from soil to surface water. In these regions no groundwater denitrification and N 2 O production is expected. Predicted N leaching (16% of the N inputs) and N 2 O emissions (0.014% of N leaching) are much less than the IPCC default leaching rate and combined emission factor for groundwater and riparian zones, respectively. - Highlights: ► Groundwater denitrification and N 2 O production was modelled at the European scale. ► In large parts of Europe no groundwater denitrification is expected. ► N leaching and N 2 O emission in Europe are much less than the IPCC default values. - European groundwater denitrification is spatially variable, and associated nitrous oxide production is much less than based on the IPCC default estimate.

Detoxification enzymes activities in deltamethrin and bendiocarb ...

African Journals Online (AJOL)

Detoxification enzymes activities in deltamethrin and bendiocarb resistant and susceptible malarial vectors ( Anopheles gambiae ) breeding in Bichi agricultural and residential sites, Kano state, Nigeria.

Fluorogenic Substrates for Visualizing Acidic Organelle Enzyme Activities.

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Fiona Karen Harlan

Full Text Available Lysosomes are acidic cytoplasmic organelles that are present in all nucleated mammalian cells and are involved in a variety of cellular processes including repair of the plasma membrane, defense against pathogens, cholesterol homeostasis, bone remodeling, metabolism, apoptosis and cell signaling. Defects in lysosomal enzyme activity have been associated with a variety of neurological diseases including Parkinson's Disease, Lysosomal Storage Diseases, Alzheimer's disease and Huntington's disease. Fluorogenic lysosomal staining probes were synthesized for labeling lysosomes and other acidic organelles in a live-cell format and were shown to be capable of monitoring lysosomal metabolic activity. The new targeted substrates were prepared from fluorescent dyes having a low pKa value for optimum fluorescence at the lower physiological pH found in lysosomes. They were modified to contain targeting groups to direct their accumulation in lysosomes as well as enzyme-cleavable functions for monitoring specific enzyme activities using a live-cell staining format. Application to the staining of cells derived from blood and skin samples of patients with Metachromatic Leukodystrophy, Krabbe and Gaucher Diseases as well as healthy human fibroblast and leukocyte control cells exhibited localization to the lysosome when compared with known lysosomal stain LysoTracker® Red DND-99 as well as with anti-LAMP1 Antibody staining. When cell metabolism was inhibited with chloroquine, staining with an esterase substrate was reduced, demonstrating that the substrates can be used to measure cell metabolism. When applied to diseased cells, the intensity of staining was reflective of lysosomal enzyme levels found in diseased cells. Substrates specific to the enzyme deficiencies in Gaucher or Krabbe disease patient cell lines exhibited reduced staining compared to that in non-diseased cells. The new lysosome-targeted fluorogenic substrates should be useful for research

Visualization of Enzyme Activities in Earthworm Biopores by In Situ Soil Zymography.

Science.gov (United States)

Razavi, Bahar S; Hoang, Duyen; Kuzyakov, Yakov

2017-01-01

Earthworms produce biopores with strongly increased microbial and enzyme activities and consequently they form microbial hotspots in soil. In extremely dynamic microhabitats and hotspots such as earthworm biopores, the in situ enzyme activities are a footprint of process rates and complex biotic interactions. The effect of earthworms on enzyme activities inside biopores, relative to earthworm-free soil, can be visualized by in situ soil zymography. Here, we describe the details of the approach and discuss its advantages and limitations. Direct zymography provides high spatial resolution for quantitative images of enzyme activities in biopores.

The influence of stormwater management practices on denitrification rates of receiving streams in an urban watershed

Science.gov (United States)

Cronenberger, M. S.; McMillan, S. K.

2011-12-01

Increasing urbanization and the subsequent disruption of floodplains has led to the need for implementing stormwater management strategies to mitigate the effects of urbanization, including soil and streambank erosion, increased export of nutrients and contaminants and decreased biotic richness. Excessive stormwater runoff due to the abundance of impervious surfaces associated with an urban landscape has led to the ubiquitous use of best management practices (BMPs) to attenuate runoff events and prevent the destructive delivery of large volumes of water to stream channels. As a result, effluent from BMPs (i.e. wetlands and wet ponds) has the potential to alter the character of the receiving stream channel and thus, key ecosystem processes such as denitrification. The purpose of this study was to determine the extent to which BMPs, in the form of constructed wetlands and wet ponds, influence in-stream denitrification rates in the urban landscape of Charlotte, NC. Four sites, two of each BMP type, were evaluated. Sediment samples were collected upstream and downstream of the BMP outflow from May-July 2011 to determine the effect of wetland discharge on in-stream nitrogen removal via denitrification. Denitrification rates were determined using the acetylene block method; water column nutrient and carbon concentrations and sediment organic matter content were also measured. Generally, wetland sites exhibited higher denitrification rates, nitrate concentrations and sediment organic matter content. Our work and others has demonstrated a significant positive correlation between nitrate concentration and denitrification rates, which is the likely driver of the higher observed rates at the wetland sites. Geomorphology was also found to be a key factor in elevated denitrification rates at sites with riffles and boulder jams. Sediment organic matter was found to be higher downstream of BMP outflows at all four sites, but demonstrated no significant relationship with

Effects of protease and non-starch polysaccharide enzyme on performance, digestive function, activity and gene expression of endogenous enzyme of broilers.

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Lin Yuan

Full Text Available Three hundred one-day-old male broiler chickens (Ross-308 were fed corn-soybean basal diets containing non-starch polysaccharide (NSP enzyme and different levels of acid protease from 1 to 42 days of age to investigate the effects of exogenous enzymes on growth performance, digestive function, activity of endogenous digestive enzymes in the pancreas and mRNA expression of pancreatic digestive enzymes. For days 1-42, compared to the control chickens, average daily feed intake (ADFI and average daily gain (ADG were significantly enhanced by the addition of NSP enzyme in combination with protease supplementation at 40 or 80 mg/kg (p<0.05. Feed-to-gain ratio (FGR was significantly improved by supplementation with NSP enzymes or NSP enzyme combined with 40 or 80 mg/kg protease compared to the control diet (p<0.05. Apparent digestibility of crude protein (ADCP was significantly enhanced by the addition of NSP enzyme or NSP enzyme combined with 40 or 80 mg/kg protease (p<0.05. Cholecystokinin (CCK level in serum was reduced by 31.39% with NSP enzyme combined with protease supplementation at 160 mg/kg (p<0.05, but the CCK level in serum was increased by 26.51% with NSP enzyme supplementation alone. After 21 days, supplementation with NSP enzyme and NSP enzyme combined with 40 or 80 mg/kg protease increased the activity of pancreatic trypsin by 74.13%, 70.66% and 42.59% (p<0.05, respectively. After 42 days, supplementation with NSP enzyme and NSP enzyme combined with 40 mg/kg protease increased the activity of pancreatic trypsin by 32.45% and 27.41%, respectively (p<0.05. However, supplementation with NSP enzyme and 80 or 160 mg/kg protease decreased the activity of pancreatic trypsin by 10.75% and 25.88%, respectively (p<0.05. The activities of pancreatic lipase and amylase were significantly higher in treated animals than they were in the control group (p<0.05. Supplementation with NSP enzyme, NSP enzyme combined with 40 or 80 mg/kg protease increased

Nanocaged enzymes with enhanced catalytic activity and increased stability against protease digestion

Science.gov (United States)

Zhao, Zhao; Fu, Jinglin; Dhakal, Soma; Johnson-Buck, Alexander; Liu, Minghui; Zhang, Ting; Woodbury, Neal W.; Liu, Yan; Walter, Nils G.; Yan, Hao

2016-01-01

Cells routinely compartmentalize enzymes for enhanced efficiency of their metabolic pathways. Here we report a general approach to construct DNA nanocaged enzymes for enhancing catalytic activity and stability. Nanocaged enzymes are realized by self-assembly into DNA nanocages with well-controlled stoichiometry and architecture that enabled a systematic study of the impact of both encapsulation and proximal polyanionic surfaces on a set of common metabolic enzymes. Activity assays at both bulk and single-molecule levels demonstrate increased substrate turnover numbers for DNA nanocage-encapsulated enzymes. Unexpectedly, we observe a significant inverse correlation between the size of a protein and its activity enhancement. This effect is consistent with a model wherein distal polyanionic surfaces of the nanocage enhance the stability of active enzyme conformations through the action of a strongly bound hydration layer. We further show that DNA nanocages protect encapsulated enzymes against proteases, demonstrating their practical utility in functional biomaterials and biotechnology. PMID:26861509

Nanocaged enzymes with enhanced catalytic activity and increased stability against protease digestion

Science.gov (United States)

Zhao, Zhao; Fu, Jinglin; Dhakal, Soma; Johnson-Buck, Alexander; Liu, Minghui; Zhang, Ting; Woodbury, Neal W.; Liu, Yan; Walter, Nils G.; Yan, Hao

2016-02-01

Cells routinely compartmentalize enzymes for enhanced efficiency of their metabolic pathways. Here we report a general approach to construct DNA nanocaged enzymes for enhancing catalytic activity and stability. Nanocaged enzymes are realized by self-assembly into DNA nanocages with well-controlled stoichiometry and architecture that enabled a systematic study of the impact of both encapsulation and proximal polyanionic surfaces on a set of common metabolic enzymes. Activity assays at both bulk and single-molecule levels demonstrate increased substrate turnover numbers for DNA nanocage-encapsulated enzymes. Unexpectedly, we observe a significant inverse correlation between the size of a protein and its activity enhancement. This effect is consistent with a model wherein distal polyanionic surfaces of the nanocage enhance the stability of active enzyme conformations through the action of a strongly bound hydration layer. We further show that DNA nanocages protect encapsulated enzymes against proteases, demonstrating their practical utility in functional biomaterials and biotechnology.

Denitrification controls in urban riparian soils: implications for reducing urban nonpoint source nitrogen pollution.

Science.gov (United States)

Li, Yangjie; Chen, Zhenlou; Lou, Huanjie; Wang, Dongqi; Deng, Huanguang; Wang, Chu

2014-09-01

The purpose of this research was to thoroughly analyze the influences of environmental factors on denitrification processes in urban riparian soils. Besides, the study was also carried out to identify whether the denitrification processes in urban riparian soils could control nonpoint source nitrogen pollution in urban areas. The denitrification rates (DR) over 1 year were measured using an acetylene inhibition technique during the incubation of intact soil cores from six urban riparian sites, which could be divided into three types according to their vegetation. The soil samples were analyzed to determine the soil organic carbon (SOC), soil total nitrogen (STN), C/N ratio, extractable NO3 (-)-N and NH4 (+)-N, pH value, soil water content (SWC), and the soil nitrification potential to evaluate which of these factors determined the final outcome of denitrification. A nitrate amendment experiment further indicated that the riparian DR was responsive to added nitrate. Although the DRs were very low (0.099 ~ 33.23 ng N2O-N g(-1) h(-1)) due to the small amount of nitrogen moving into the urban riparian zone, the spatial and temporal patterns of denitrification differed significantly. The extractable NO3 (-)-N proved to be the dominant factor influencing the spatial distribution of denitrification, whereas the soil temperature was a determinant of the seasonal DR variation. The six riparian sites could also be divided into two types (a nitrate-abundant and a nitrate-stressed riparian system) according to the soil NO3 (-)-N concentration. The DR in nitrate-abundant riparian systems was significantly higher than that in the nitrate-stressed riparian systems. The DR in riparian zones that were covered with bushes and had adjacent cropland was higher than in grass-covered riparian sites. Furthermore, the riparian DR decreased with soil depth, which was mainly attributed to the concentrated nitrate in surface soils. The DR was not associated with the SOC, STN, C/N ratio, and

Simultaneous heterotrophic and sulfur-oxidizing autotrophic denitrification process for drinking water treatment: control of sulfate production.

Science.gov (United States)

Sahinkaya, Erkan; Dursun, Nesrin; Kilic, Adem; Demirel, Sevgi; Uyanik, Sinan; Cinar, Ozer

2011-12-15

A long-term performance of a packed-bed bioreactor containing sulfur and limestone was evaluated for the denitrification of drinking water. Autotrophic denitrification rate was limited by the slow dissolution rate of sulfur and limestone. Dissolution of limestone for alkalinity supplementation increased hardness due to release of Ca(2+). Sulfate production is the main disadvantage of the sulfur autotrophic denitrification process. The effluent sulfate concentration was reduced to values below drinking water guidelines by stimulating the simultaneous heterotrophic and autotrophic denitrification with methanol supplementation. Complete removal of 75 mg/L NO(3)-N with effluent sulfate concentration of around 225 mg/L was achieved when methanol was supplemented at methanol/NO(3)-N ratio of 1.67 (mg/mg), which was much lower than the theoretical value of 2.47 for heterotrophic denitrification. Batch studies showed that sulfur-based autotrophic NO(2)-N reduction rate was around three times lower than the reduction rate of NO(3)-N, which led to NO(2)-N accumulation at high loadings. Copyright © 2011 Elsevier Ltd. All rights reserved.

Activity of certain enzymes in cadmium-poisoned chicks

Energy Technology Data Exchange (ETDEWEB)

Kench, J E; Gubb, P J.D.

1970-01-01

Activities of a number of enzymes in the liver and other tissues of newly hatched cadmium poisoned chicks have been compared with those of normal controls before and after incubation with Cd/sup +2/ at a concentration similar to that present in vivo. Concentrations of Cd/sup +2/ in the various cellular fractions were determined, after wet oxidation, by atomic absorption spectrophotometry. Interaction of Cd/sup +2/ with enzymes may provide information on the localization of enzymes within mitochondria and other cellular structures. 7 references.

Development Of Enzyme Digestive Activity Of Blue Crab Portunus Pelagicus Larvae

OpenAIRE

Nikhlani, Andi; Sukarti, Komsanah

2017-01-01

Seed production continuity of Portunus pelagicus larvae had been conducted but the results were still un-consistent Digestive activity was known to be associated with the type of feed consumed by larvae. Amylase, lipase, and trypsin enzymes were used as a biological indicators to measure the digestion of feed. The aim of this study was to describe the activity of digestive enzymes in blue swimming crab larvae. Digestive enzyme activity data obtained was presented in graphical form and anal...

Potential denitrification and nitrous oxide production in the sediments of the Seine River Drainage Network (France).

Science.gov (United States)

Garnier, Josette A; Mounier, Emmanuelle M; Laverman, Anniet M; Billen, Gilles F

2010-01-01

To investigate bottom sediment denitrification at the scale of the Seine drainage network, a semi-potential denitrification assay was used in which river sediments (and riparian soils) were incubated for a few hours under anaerobic conditions with non limiting nitrate concentrations. This method allowed the nitrous oxide (N(2)O) concentration in the headspace, as well as the nitrate, nitrite, and ammonium concentrations to be determined during incubation. The rates at which nitrate decreased and N(2)O increased were then used to assess the potential denitrification activity and associated N(2)O production in the Seine River Basin. We observed a longitudinal pattern characterized by a significant increase of the potential rate of denitrification from upstream sectors to large downstream rivers (orders 7-8), from approximately 3.3 to 9.1 microg NO(3)(-)-N g(-1) h(-1), respectively, while the N(2)O production rates was the highest both in headwaters and in large order rivers (0.14 and 0.09 N(2)O-N g(-1) h(-1), respectively) and significantly lower in the intermediate sectors (0.01 and 0.03 N(2)O-N g(-1) h(-1)). Consequently, the ratio N(2)O production:NO(3) reduction was found to reach 5% in headstreams, whereas it averaged 1.2% in the rest of the drainage network, an intermediate percentage being found for the riparian soils. Finally, the ignition loss of sediments, together with other redundant variables (particulate organic carbon content: g C 100 g(-1) dry weight [dw], moisture: g water 100 g(-1) dw, sediment size production.

Pathogenicity and cell wall-degrading enzyme activities of some ...

African Journals Online (AJOL)

Dr. J. T. Ekanem

2005-12-17

Dec 17, 2005 ... be attributed to the activities of these cell wall degrading enzymes. Keywords: Cowpea ... bacteria have long been known to produce enzymes capable of ... Inoculated seeds were sown in small plastic pots filled with steam- ...

Plant carbohydrate binding module enhances activity of hybrid microbial cellulase enzyme

Directory of Open Access Journals (Sweden)

Caitlin Siobhan Byrt

2012-11-01

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

The efficiency of a membrane bioreactor in drinking water denitrification

Directory of Open Access Journals (Sweden)

Petrovič Aleksandra

2015-01-01

Full Text Available The membrane bioreactor (MBR system was investigated regarding its nitrate removal capacity from drinking water. The performance of a pilot-scale MBR was tested, depending on the operational parameters, using sucrose as a carbon source. Drinking water from the source was introduced into the reactor in order to study the influence of flow-rate on the nitrate removal and denitrification efficiency of drinking water. The content of the nitrate was around 70 mg/L and the C/N ratio was 3:1. Nitrate removal efficiencies above 90% were obtained by flow-rates lower than 4.8 L/h. The specific denitrification rates varied between 0.02 and 0.16 g/L NO3/ (g/L MLSS•d. The efficiencies and nitrate removal were noticeably affected by the flow-rate and hydraulic retention times. At the maximum flow-rate of 10.2 L/h still 68% of the nitrate had been removed, whilst the highest specific denitrification rate was achieved at 0.2738 g/L NO3/ (g/L MLSS•d. The maximum reactor removal capacity was calculated at 8.75 g NO3/m3•h.

Combined denitrification and phosphorus removal in a biofilter

DEFF Research Database (Denmark)

Falkentoft, Christina Maria; Harremoes, Poul; Mosbæk, Hans

2000-01-01

A lab-scale biofilter was run continuously for 11/2 years for combined denitrification and phosphorus removal. Alternation between anaerobic and anoxic (nitrate) conditions was used to obtain an enriched culture of denitrifying, phosphate accumulating organisms. Batch experiments were performed...

Coastal versus open-ocean denitrification in the Arabian Sea

Digital Repository Service at National Institute of Oceanography (India)

Naqvi, S.W.A.; Naik, H.; Pratihary, A.K.; DeSouza, W.; Narvekar, P.V.; Jayakumar, D.A.; Devol, A.H.; Yoshinari, T.; Saino, T.

through advection from the south. The waters Published by Copernicus GmbH on behalf of the European Geosciences Union. 622 S. W. A. Naqvi et al.: Arabian Sea denitrification 60 E 70 E 10 N 20 N 3204 3939 3201 TN039/18 CATS � � (a) o o o 0 500 1000 1500....: Arabian Sea denitrification 625 150 100 50 Temperature (oC) 100 50 O2 (M) Distance from coast (km) 150 100 50 Salinity G2 CATS G9 �P�P 150 100 50 150 100 50 150 NO2- (M) 150 100 50 NO3-(M) N2O (nM) 150 100 50 NH4+(M) 80 60 40 20 0 Distance from coast (km...

Reveal the response of enzyme activities to heavy metals through in situ zymography.

Science.gov (United States)

Duan, Chengjiao; Fang, Linchuan; Yang, Congli; Chen, Weibin; Cui, Yongxing; Li, Shiqing

2018-07-30

Enzymes in the soil are vital for assessing heavy metal soil pollution. Although the presence of heavy metals is thought to change the soil enzyme system, the distribution of enzyme activities in heavy metal polluted-soil is still unknown. For the first time, using soil zymography, we analyzed the distribution of enzyme activities of alfalfa rhizosphere and soil surface in the metal-contaminated soil. The results showed that the growth of alfalfa was significantly inhibited, and an impact that was most pronounced in seedling biomass and chlorophyll content. Catalase activity (CAT) in alfalfa decreased with increasing heavy metal concentrations, while malondialdehyde (MDA) content continually increased. The distribution of enzyme activities showed that both phosphatase and β-glucosidase activities were associated with the roots and were rarely distributed throughout the soil. In addition, the total hotspot areas of enzyme activities were the highest in extremely heavy pollution soil. The hotspot areas of phosphatase were 3.4%, 1.5% and 7.1% under none, moderate and extremely heavy pollution treatment, respectively, but increased from 0.1% to 0.9% for β-glucosidase with the increasing pollution levels. Compared with the traditional method of enzyme activities, zymography can directly and accurately reflect the distribution and extent of enzyme activity in heavy metals polluted soil. The results provide an efficient research method for exploring the interaction between enzyme activities and plant rhizosphere. Copyright © 2018 Elsevier Inc. All rights reserved.

Glacial-interglacial changes and Holocene variations in Arabian Sea denitrification

Science.gov (United States)

Gaye, Birgit; Böll, Anna; Segschneider, Joachim; Burdanowitz, Nicole; Emeis, Kay-Christian; Ramaswamy, Venkitasubramani; Lahajnar, Niko; Lückge, Andreas; Rixen, Tim

2018-01-01

At present, the Arabian Sea has a permanent oxygen minimum zone (OMZ) at water depths between about 100 and 1200 m. Active denitrification in the upper part of the OMZ is recorded by enhanced δ15N values in the sediments. Sediment cores show a δ15N increase during the middle and late Holocene, which is contrary to the trend in the other two regions of water column denitrification in the eastern tropical North and South Pacific. We calculated composite sea surface temperature (SST) and δ15N ratios in time slices of 1000 years of the last 25 kyr to better understand the reasons for the establishment of the Arabian Sea OMZ and its response to changes in the Asian monsoon system. Low δ15N values of 4-7 ‰ during the last glacial maximum (LGM) and stadials (Younger Dryas and Heinrich events) suggest that denitrification was inactive or weak during Pleistocene cold phases, while warm interstadials (ISs) had elevated δ15N. Fast changes in upwelling intensities and OMZ ventilation from the Antarctic were responsible for these strong millennial-scale variations during the glacial. During the entire Holocene δ15N values > 6 ‰ indicate a relatively stable OMZ with enhanced denitrification. The OMZ develops parallel to the strengthening of the SW monsoon and monsoonal upwelling after the LGM. Despite the relatively stable climatic conditions of the Holocene, the δ15N records show regionally different trends in the Arabian Sea. In the upwelling areas in the western part of the basin, δ15N values are lower during the mid-Holocene (4.2-8.2 ka BP) compared to the late Holocene ( ventilation of the OMZ during the period of the most intense southwest monsoonal upwelling. In contrast, δ15N values in the northern and eastern Arabian Sea rose during the last 8 kyr. The displacement of the core of the OMZ from the region of maximum productivity in the western Arabian Sea to its present position in the northeast was established during the middle and late Holocene. This was

Glacial–interglacial changes and Holocene variations in Arabian Sea denitrification

Directory of Open Access Journals (Sweden)

B. Gaye

2018-01-01

Full Text Available At present, the Arabian Sea has a permanent oxygen minimum zone (OMZ at water depths between about 100 and 1200 m. Active denitrification in the upper part of the OMZ is recorded by enhanced δ15N values in the sediments. Sediment cores show a δ15N increase during the middle and late Holocene, which is contrary to the trend in the other two regions of water column denitrification in the eastern tropical North and South Pacific. We calculated composite sea surface temperature (SST and δ15N ratios in time slices of 1000 years of the last 25 kyr to better understand the reasons for the establishment of the Arabian Sea OMZ and its response to changes in the Asian monsoon system. Low δ15N values of 4–7 ‰ during the last glacial maximum (LGM and stadials (Younger Dryas and Heinrich events suggest that denitrification was inactive or weak during Pleistocene cold phases, while warm interstadials (ISs had elevated δ15N. Fast changes in upwelling intensities and OMZ ventilation from the Antarctic were responsible for these strong millennial-scale variations during the glacial. During the entire Holocene δ15N values  >  6 ‰ indicate a relatively stable OMZ with enhanced denitrification. The OMZ develops parallel to the strengthening of the SW monsoon and monsoonal upwelling after the LGM. Despite the relatively stable climatic conditions of the Holocene, the δ15N records show regionally different trends in the Arabian Sea. In the upwelling areas in the western part of the basin, δ15N values are lower during the mid-Holocene (4.2–8.2 ka BP compared to the late Holocene ( <  4.2 ka BP due to stronger ventilation of the OMZ during the period of the most intense southwest monsoonal upwelling. In contrast, δ15N values in the northern and eastern Arabian Sea rose during the last 8 kyr. The displacement of the core of the OMZ from the region of maximum productivity in the western Arabian Sea to its present position

Nitrate removal from groundwater by cooperating heterotrophic with autotrophic denitrification in a biofilm-electrode reactor

Energy Technology Data Exchange (ETDEWEB)

Zhao, Yingxin [School of Water Resources and Environment, China University of Geosciences, Beijing 100083 (China); Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba 3058572 (Japan); Feng, Chuanping, E-mail: fengchuangping@gmail.com [School of Water Resources and Environment, China University of Geosciences, Beijing 100083 (China); Wang, Qinghong; Yang, Yingnan; Zhang, Zhenya; Sugiura, Norio [Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba 3058572 (Japan)

2011-09-15

Highlights: {yields} Intensified biofilm-electrode reactor using cooperative denitrification is developed. {yields} IBER combines heterotrophic and autotrophic denitrification. {yields} CO{sub 2} formed by heterotrophic denitrification is used by autotrophic bacteria. {yields} Optimum running conditions are C/N = 0.75, HRT = 8 h, and I = 40 mA. {yields} A novel degradation mechanism for cooperating denitrification process is proposed. - Abstract: An intensified biofilm-electrode reactor (IBER) combining heterotrophic and autotrophic denitrification was developed for treatment of nitrate contaminated groundwater. The reactor was evaluated with synthetic groundwater (NO{sub 3}{sup -}N50 mg L{sup -1}) under different hydraulic retention times (HRTs), carbon to nitrogen ratios (C/N) and electric currents (I). The experimental results demonstrate that high nitrate and nitrite removal efficiency (100%) were achieved at C/N = 1, HRT = 8 h, and I = 10 mA. C/N ratios were reduced from 1 to 0.5 and the applied electric current was changed from 10 to 100 mA, showing that the optimum running condition was C/N = 0.75 and I = 40 mA, under which over 97% of NO{sub 3}{sup -}N was removed and organic carbon (methanol) was completely consumed in treated water. Simultaneously, the denitrification mechanism in this system was analyzed through pH variation in effluent. The CO{sub 2} produced from the anode acted as a good pH buffer, automatically controlling pH in the reaction zone. The intensified biofilm-electrode reactor developed in the study was effective for the treatment of groundwater polluted by nitrate.

DNA-directed control of enzyme-inhibitor complex formation: a modular approach to reversibly switch enzyme activity

NARCIS (Netherlands)

Janssen, B.M.G.; Engelen, W.; Merkx, M.

2015-01-01

DNA-templated reversible assembly of an enzyme–inhibitor complex is presented as a new and highly modular approach to control enzyme activity. TEM1-ß-lactamase and its inhibitor protein BLIP were conjugated to different oligonucleotides, resulting in enzyme inhibition in the presence of template

Micropollutant degradation via extracted native enzymes from activated sludge.

Science.gov (United States)

Krah, Daniel; Ghattas, Ann-Kathrin; Wick, Arne; Bröder, Kathrin; Ternes, Thomas A

2016-05-15

A procedure was developed to assess the biodegradation of micropollutants in cell-free lysates produced from activated sludge of a municipal wastewater treatment plant (WWTP). This proof-of-principle provides the basis for further investigations of micropollutant biodegradation via native enzymes in a solution of reduced complexity, facilitating downstream protein analysis. Differently produced lysates, containing a variety of native enzymes, showed significant enzymatic activities of acid phosphatase, β-galactosidase and β-glucuronidase in conventional colorimetric enzyme assays, whereas heat-deactivated controls did not. To determine the enzymatic activity towards micropollutants, 20 compounds were spiked to the cell-free lysates under aerobic conditions and were monitored via LC-ESI-MS/MS. The micropollutants were selected to span a wide range of different biodegradabilities in conventional activated sludge treatment via distinct primary degradation reactions. Of the 20 spiked micropollutants, 18 could be degraded by intact sludge under assay conditions, while six showed reproducible degradation in the lysates compared to the heat-deactivated negative controls: acetaminophen, N-acetyl-sulfamethoxazole (acetyl-SMX), atenolol, bezafibrate, erythromycin and 10,11-dihydro-10-hydroxycarbamazepine (10-OH-CBZ). The primary biotransformation of the first four compounds can be attributed to amide hydrolysis. However, the observed biotransformations in the lysates were differently influenced by experimental parameters such as sludge pre-treatment and the addition of ammonium sulfate or peptidase inhibitors, suggesting that different hydrolase enzymes were involved in the primary degradation, among them possibly peptidases. Furthermore, the transformation of 10-OH-CBZ to 9-CA-ADIN was caused by a biologically-mediated oxidation, which indicates that in addition to hydrolases further enzyme classes (probably oxidoreductases) are present in the native lysates. Although the

Denitrification potential and its relation to organic carbon quality in three coastal wetland soils

International Nuclear Information System (INIS)

Dodla, Syam K.; Wang, Jim J.; DeLaune, Ron D.; Cook, Robert L.

2008-01-01

Capacity of a wetland to remove nitrate through denitrification is controlled by its physico-chemical and biological characteristics. Understanding these characteristics will help better to guide beneficial use of wetlands in processing nitrate. This study was conducted to determine the relationship between soil organic carbon (SOC) quality and denitrification rate in Louisiana coastal wetlands. Composite soil samples of different depths were collected from three different wetlands along a salinity gradient, namely, bottomland forest swamp (FS), freshwater marsh (FM), and saline marsh (SM) located in the Barataria Basin estuary. Potential denitrification rate (PDR) was measured by acetylene inhibition method and distribution of carbon (C) moieties in organic C was determined by 13 C solid-state NMR. Of the three wetlands, the FM soil profile exhibited the highest PDR on both unit weight and unit volume basis as compared to FS and SM. The FM also tended to yield higher amount of N 2 O as compared to the FS and SM especially at earlier stages of denitrification, suggesting incomplete reduction of NO 3 - at FM and potential for emission of N 2 O. Saline marsh soil profile had the lowest PDR on the unit volume basis. Increasing incubation concentration from 2 to 10 mg NO 3 - -N L -1 increased PDR by 2 to 6 fold with the highest increase in the top horizons of FS and SM soils. Regression analysis showed that across these three wetland systems, organic C has significant effect in regulating PDR. Of the compositional C moieties, polysaccharides positively influenced denitrification rate whereas phenolics (likely phenolic adehydes and ketonics) negatively affected denitrification rate in these wetland soils. These results could have significant implication in integrated assessment and management of wetlands for treating nutrient-rich biosolids and wastewaters, non-point source agricultural runoff, and nitrate found in the diverted Mississippi River water used for coastal

Effect of dissolved oxygen on biological denitrification using biodegradable plastic as the carbon source

Science.gov (United States)

Zhang, Xucai; Zhang, Jianmei

2018-02-01

Biological denitrification is currently a common approach to remove nitrate from wastewater. This study was conducted to evaluate the influence of dissolved oxygen on denitrification in wastewater treatment using biodegradable plastic as carbon source by designing the aerated, anoxic, and low-oxygen experimental treatment groups. The results showed that the removal rates of nitrate in anoxic and low-oxygen groups were 30.6 g NO3 --Nm-3 d-1 and 30.8 g NO3 --N m-3 d-1 at 83 h, respectively, both of which were higher than that of the aerated group. There was no significant difference between the anoxic and low-oxygen treatment groups for the nitrate removal. Additional, the nitrite accumulated during the experiments, and the nitrite concentrations in anoxic and aerated groups were lower than those in low-oxygen group. No nitrite was detected in all groups at the end of the experiments. These findings indicated that dissolved oxygen has important influence on denitrification, and anoxic and low-oxygen conditions can support completely denitrification when using BP as carbon source in nitrate-polluted wastewater treatment.

Rising Sludge in Secondary Settlers Due to Denitrification

DEFF Research Database (Denmark)

Henze, Mogens; Dupont, Rene; Grau, Peter

1993-01-01

High suspended solids concentrations in settler effluents can be caused by rising sludge, which is the effect of flotation of solids by nitrogen gas resulting from biological denitrification. Many factors influence the nitrogen gas bubble evolution. The most important factor is the rate...

Acid Fermentation Process Combined with Post Denitrification for the Treatment of Primary Sludge and Wastewater with High Strength Nitrate

Directory of Open Access Journals (Sweden)

Allen Kurniawan

2016-03-01

Full Text Available In this study, an anaerobic baffled reactor (ABR, combined with a post denitrification process, was applied to treat primary sludge from a municipal wastewater treatment plant and wastewater with a high concentration of nitrate. The production of volatile fatty acids (VFAs was maximized with a short hydraulic retention time in the acid fermentation of the ABR process, and then the produced VFAs were supplied as an external carbon source for the post denitrification process. The laboratory scale experiment was operated for 160 days to evaluate the VFAs’ production rate, sludge reduction in the ABR type-acid fermentation process, and the specific denitrification rate of the post denitrification process. As results, the overall removal rate of total chemical oxygen demand (TCOD, total suspended solids (TSS, and total nitrogen (TN were found to be 97%, 92%, 73%, respectively, when considering the influent into ABR type-acid fermentation and effluent from post denitrification. We observed the specific VFAs production rate of 0.074 gVFAs/gVSS/day for the ABR type-acid fermentation, and an average specific denitrification rate of 0.166 gNO3−-N/gVSS/day for the post denitrification. Consequently, we observed that a high production of VFAs from a primary sludge, using application of the ABR type acid fermentation process and the produced VFAs were then successfully utilized as an external carbon source for the post denitrification process, with a high removal rate of nitrogen.

Activation of lysosomal enzymes and tumour regression caused by irradiation and steroid hormones

International Nuclear Information System (INIS)

Ball, A.; Barratt, G.M.; Wills, E.D.

1982-01-01

The lysosomal enzyme activity and membrane permeability of mouse C3H mammary tumours has been studied using quantitative cytochemical methods following irradiation of the tumours with doses of 1500, 3500 or 6000 rad ν rays. No change in the lysosomal enzyme activity was observed immediately after irradiation, but increased enzyme activity and increased membrane permeability were observed 24 hr after irradiation with doses of 3500 or 6000 rad. Twenty-four hours after injection of prednisolone there was a marked increase of lysosomal membrane permeability and enzyme activity, and injection of prednisolone soon after irradiation enhanced the effect of irradiation. After a dose of 6000 rad and prednisolone, the lysosomal membrane permeability increased to 191% of the control and the enzyme activity to 326% of the value of the control tumours. Measurement of tumour size after irradiation or after a combined treatment with irradiation and prednisolone showed that a close correlation exists between tumour regression and lysosomal enzyme activity. The experiments support the view that lysosomal enzymes play an important role in tumour regression following irradiation. (author)

[Identification and Nitrogen Removal Characteristics of a Heterotrophic Nitrification-Aerobic Denitrification Strain Isolated from Marine Environment].

Science.gov (United States)

Sun, Qing-hua; Yu, De-shuang; Zhang, Pei-yu; Lin, Xue-zheng; Li, Jin

2016-02-15

A heterotrophic nitrification-aerobic denitrification strain named y5 was isolated from marine environment by traditional microbial isolation method using seawater as medium. It was identified as Klebsiella sp. based on the morphological, physiological and 16S rRNA sequence analysis. The experiment results showed that the optimal carbon resource was sodium citrate; the optimal pH was 7.0; and the optimal C/N was 17. The strain could use NH4Cl, NaNO2 and KNO3 as sole nitrogen source, and the removal efficiencies were77.07%, 64.14% and 100% after 36 hours, respectively. The removal efficiency reached 100% after 36 hours in the coexistence of NH4Cl, NaNO2 and KNO3. The results showed that the strain y5 had independent and efficient heterotrophic nitrification and aerobic denitrification activities in high salt wastewater.

Hfq stimulates the activity of the CCA-adding enzyme

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Betat Heike

2007-10-01

Full Text Available Abstract Background The bacterial Sm-like protein Hfq is known as an important regulator involved in many reactions of RNA metabolism. A prominent function of Hfq is the stimulation of RNA polyadenylation catalyzed by E. coli poly(A polymerase I (PAP. As a member of the nucleotidyltransferase superfamily, this enzyme shares a high sequence similarity with an other representative of this family, the tRNA nucleotidyltransferase that synthesizes the 3'-terminal sequence C-C-A to all tRNAs (CCA-adding enzyme. Therefore, it was assumed that Hfq might not only influence the poly(A polymerase in its specific activity, but also other, similar enzymes like the CCA-adding enzyme. Results Based on the close evolutionary relation of these two nucleotidyltransferases, it was tested whether Hfq is a specific modulator acting exclusively on PAP or whether it also influences the activity of the CCA-adding enzyme. The obtained data indicate that the reaction catalyzed by this enzyme is substantially accelerated in the presence of Hfq. Furthermore, Hfq binds specifically to tRNA transcripts, which seems to be the prerequisite for the observed effect on CCA-addition. Conclusion The increase of the CCA-addition in the presence of Hfq suggests that this protein acts as a stimulating factor not only for PAP, but also for the CCA-adding enzyme. In both cases, Hfq interacts with RNA substrates, while a direct binding to the corresponding enzymes was not demonstrated up to now (although experimental data indicate a possible interaction of PAP and Hfq. So far, the basic principle of these stimulatory effects is not clear yet. In case of the CCA-adding enzyme, however, the presented data indicate that the complex between Hfq and tRNA substrate might enhance the product release from the enzyme.

Direct contribution of clams (Ruditapes philippinarum) to benthic fluxes, nitrification, denitrification and nitrous oxide emission in a farmed sediment

Science.gov (United States)

Welsh, David T.; Nizzoli, Daniele; Fano, Elisa A.; Viaroli, Pierluigi

2015-03-01

The influence of the manila clam (Ruditapes philippinarum) on N-cycle processes, and oxygen and nutrient fluxes in a farmed sediment was investigated using a multiple core incubation approach and parallel incubations of individual clams. Clam population/biomass density varied ∼8-fold between cores and all sediment-water column solute (O2. N2, N2O, NH4+, NOX and DIN) fluxes and benthic process (N-regeneration, nitrification and denitrification) rates were strongly and significantly correlated with clam density/biomass. Isolated clams exhibited high rates of respiration, N-excretion, nitrification and denitrification of 2050 ± 70, 395 ± 49, 201 ± 42 and 235 ± 40 nmol individual-1 h-1, respectively. The direct contribution of the clams and their associated microbiota to benthic processes was estimated by multiplying the per individual rates by the number of clams in each incubated core. The clams on average directly accounted for 64-133% of total rates of sediment oxygen demand, N-regeneration, nitrification and denitrification, indicating that they regulated processes primarily through their own metabolic activity and that of bacteria that colonise them. Clams and the farmed sediments were significant sources of the greenhouse gas N2O, but this was primarily due to their high nitrification and denitrification rates, rather than high specific N2O yields, as N2O emissions represented farmed sediments had a high denitrification efficiency of 67 ± 10%, but this ecosystem service came at the environmental cost of increased N-regeneration and N2O emission rates. The measured N2O emissions indicate that bivalve aquaculture may be a significant source of N2O. It is therefore recommended that N2O emissions should be included in the impact assessments of current and future bivalve-farming projects.

Carotenoid-cleavage activities of crude enzymes from Pandanous amryllifolius.

Science.gov (United States)

Ningrum, Andriati; Schreiner, Matthias

2014-11-01

Carotenoid degradation products, known as norisoprenoids, are aroma-impact compounds in several plants. Pandan wangi is a common name of the shrub Pandanus amaryllifolius. The genus name 'Pandanus' is derived from the Indonesian name of the tree, pandan. In Indonesia, the leaves from the plant are used for several purposes, e.g., as natural colorants and flavor, and as traditional treatments. The aim of this study was to determine the cleavage of β-carotene and β-apo-8'-carotenal by carotenoid-cleavage enzymes isolated from pandan leaves, to investigate dependencies of the enzymatic activities on temperature and pH, to determine the enzymatic reaction products by using Headspace Solid Phase Microextraction Gas Chromatography/Mass Spectrophotometry (HS-SPME GC/MS), and to investigate the influence of heat treatment and addition of crude enzyme on formation of norisoprenoids. Crude enzymes from pandan leaves showed higher activity against β-carotene than β-apo-8'-carotenal. The optimum temperature of crude enzymes was 70°, while the optimum pH value was 6. We identified β-ionone as the major volatile reaction product from the incubations of two different carotenoid substrates, β-carotene and β-apo-8'-carotenal. Several treatments, e.g., heat treatment and addition of crude enzymes in pandan leaves contributed to the norisoprenoid content. Our findings revealed that the crude enzymes from pandan leaves with carotenoid-cleavage activity might provide a potential application, especially for biocatalysis, in natural-flavor industry. Copyright © 2014 Verlag Helvetica Chimica Acta AG, Zürich.

Enzyme activities by indicator of quality in organic soil

Science.gov (United States)

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

2016-04-01

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

High rates of denitrification and nitrous oxide emission in arid biological soil crusts from the Sultanate of Oman

DEFF Research Database (Denmark)

Abed, Raeid M M; Lam, Phyllis; De Beer, Dirk

2013-01-01

Using a combination of process rate determination, microsensor profiling and molecular techniques, we demonstrated that denitrification, and not anaerobic ammonium oxidation (anammox), is the major nitrogen loss process in biological soil crusts from Oman. Potential denitrification rates were 584...... that nitrogen loss via denitrification is a dominant process in crusts from Oman, which leads to N 2 O gas emission and potentially reduces desert soil fertility....

Indian Monsoon and denitrification change in the Laxmi Basin (IODP Exp. 355 Site U1456) of the Eastern Arabian Sea during the last 800 kyrs

Science.gov (United States)

Kim, J. E.; Khim, B. K.; Ikehara, M.; Lee, J.

2017-12-01

The Arabian Sea is a famous site for the basin-wide denitrification in the globe. The Western Arabian Sea has been acknowledged by its upwelling-induced denitrification related to the Indian Monsoon system (Altabet et al., 1999). It was recently reported that the denitrification in the Eastern Arabian Sea (IODP Exp. 355 Site U1456) has been persistent and consistent during the mid-Pleistocene as reflected in the bulk sediment δ15N values (Tripathi et al., 2017). Based on the age model reconstructed by δ18O stratigraphy of planktonic foraminifera (Globigerinoides ruber) together with shipboard biostratigraphic and paleomagnetic data at Site U1456 drilled in the Laxmi Basin of the Eastern Arabian Sea, the glacial-interglacial fluctuations of denitrification in association with the development of oxygen minimum zone (OMZ) were resolved in the context of Indian Monsoon activity. One of striking features in the Eastern Arabian Sea is that the δ15N values of bulk sediment show clear and consistent denitrification with minimum δ15N values exceeding 6‰ even during glacial periods, when its western counterpart experienced a temporal collapse of OMZ and denitrification. The Eastern Arabian Sea is fed not only by the upwelling-induced productivity in the western margin during the summer monsoon but also by the high productivity during the winter monsoon, both of which maintain the increased productivity affecting the OMZ through the consumption of dissolved oxygen by the degradation of sinking organic particles. The Eastern Arabian Sea is further influenced by the clockwise surface currents, intermediate water ventilation change by the blockage of Antarctic Intermediate Water, limited inflow from the Red Sea/Persian Gulf, and the freshwater salinity stratification due to nearby riverine discharges, all of which make the denitrification process more complicated than the Western Arabian Sea. Nonetheless, the glacial-interglacial denitrification change in the Eastern

Effect of diffusion on enzyme activity in a microreactor

NARCIS (Netherlands)

Swarts, J.W.; Kolfschoten, R.C.; Jansen, M.C.A.A.; Janssen, A.E.M.; Boom, R.M.

2010-01-01

To establish general rules for setting up an enzyme microreactor system, we studied the effect of diffusion on enzyme activity in a microreactor. As a model system we used the hydrolysis of ortho-nitrophenyl-ß-d-galactopyranoside by ß-galactosidase from Kluyveromyces lactis. We found that the

Differentiation between activity of digestive enzymes of Brachionus calyciflorus and extracellular enzymes of its epizooic bacteria

Directory of Open Access Journals (Sweden)

Wilko H. AHLRICHS

2009-08-01

Full Text Available The rotifer Brachionus calyciflorus was examined by scanning electron microscopy (SEM for surface-attached, i.e. epizootic, bacteria to ascertain their specific localization and thus find out if we could discern between rotifer and bacterial enzyme activity. The lorica of B. calyciflorus was colonized by one distinct type of bacteria, which originated from the algal culture used for rotifer feeding. The corona, posterior epidermis and foot of all inspected individuals were always without attached bacteria. The density of the attached bacteria was higher with the increasing age of B. calyciflorus: while young individuals were colonized by ~ tens of bacterial cells, older ones had on average hundreds to thousands of attached bacteria. We hypothesize that epizooic bacteria may produce the ectoenzymes phosphatases and β-N-acetylhexosaminidases on the lorica, but not on the corona of B. calyciflorus. Since enzyme activities of epizooic bacteria may influence the values and interpretation of bulk rotifer enzyme activities, we should take the bacterial contribution into account.

Enzyme activities in reclaimed coal mine spoils and soils

Energy Technology Data Exchange (ETDEWEB)

Fresquez, P R; Aldon, E F; Lindemann, W C

1987-11-01

The segregation and stockpiling of topsoil material may reduce enzymatic activities that may hinder normal nutrient cycling processes in reclaimed minelands. The effects of topsoiling and reclamation age on dehydrogenase, nitrogenase, phosphatase, arylsulphatase, amylase, cellulase, invertase and urease activities were evaluated on three reclaimed non-top-soiled and five reclaimed topsoiled areas and compared with an indisturbed reference soil. Three months after topsoiling and revegetation, activities of the enzymes in the reclaimed areas, with the exception of dehydrogenase, were statistically equal to activities of the undisturbed soil. Most enzymes, including dehydrogenase, peaked in the next 1 or 2 years after reclamation with topsoiling and declined thereafter. A 4-year-old topsoiled site (revegetated in 1978) was statistically similar to the undisturbed soil. Amylase activity, however, was significantly lower after the fourth year compared to the undisturbed soil. The non-topsoiled areas, even after 6, 7 and 8 years, appeared to have lower enzyme activities than the younger topsoiled areas or the undisturbed soil. This trend was supported by the finding that the 4-year-old topsoiled site was more enzymatically similar to the undisturbed soil than was the 8-year-old non-topsoiled site (revegetated in 1974). The low enzyme acitivities found in the non-topsoiled areas may be a result of their adverse chemical and physical properties, as well as the low diversity of microorganisms. These studies demonstrate the value of topsoil use for early establishment of soil processes in reclaimed areas. 3 figs., 19 refs., 8 tabs.

In Situ Denitrification and Biological Nitrogen Fixation Under Enhanced Atmospheric Reactive Nitrogen Deposition in UK Peatlands

Science.gov (United States)

Ullah, Sami; Saiz Val, Ernesto; Sgouridis, Fotis; Peichl, Matthias; Nilsson, Mats

2017-04-01

Dinitrogen (N2) and nitrous oxide (N2O) losses due to denitrification and biological N2 fixation (BNF) are the most uncertain components of the nitrogen (N) cycle in peatlands under enhanced atmospheric reactive nitrogen (Nr) deposition. This uncertainty hampers our ability to assess the contribution of denitrification to the removal of biologically fixed and/or atmospherically deposited Nr in peatlands. This uncertainty emanates from the difficulty in measuring in situ soil N2 and N2O production and consumption in peatlands. In situ denitrification and its contribution to total N2O flux was measured monthly between April 2013 and October 2014 in peatlands in two UK catchments. An adapted 15N-Gas Flux method1 with low level addition of 15N tracer (0.03 ± 0.005 kg 15N ha-1) was used to measure denitrification and its contribution to net N2O production (DN2O/TN2O). BNF was measured in situ through incubation of selected sphagnum species under 15N2 gas tracer. Denitrification2 varied temporally and averaged 8 kg N-N2 ha-1 y-1. The contribution of denitrification was about 48% to total N2O flux3 of 0.05 kg N ha-1 y-1. Soil moisture, temperature, ecosystem respiration, pH and mineral N content mainly regulated the flux of N2 and N2O. Preliminary results showed suppression of BNF, which was 1.8 to 7 times lower in peatland mosses exposed to ˜15 to 20 kg N ha-1 y-1 Nr deposition in the UK than in peatland mosses in northern Sweden with background Nr deposition. Overall, the contribution of denitrification to Nr removal in the selected peatlands was ˜50% of the annual Nr deposition rates, making these ecosystems vulnerable to chronic N saturation. These results point to a need for a more comprehensive annual BNF measurement to more accurately account for total Nr input into peatlands and its atmospheric loss due to denitrification. References Sgouridis F, Stott A & Ullah S, 2016. Application of the 15N-Gas Flux method for measuring in situ N2 and N2O fluxes due to

Phytobiotic Utilization as Feed Additive in Feed for Pancreatic Enzyme Activity of Broiler Chicken

Directory of Open Access Journals (Sweden)

Sri Purwanti

2015-09-01

Full Text Available This research was conducted to evaluate the effect of turmeric water extract, garlic and combination turmeric and garlic as a feed additive in the broiler diet on pancreatic enzyme activity of broiler chicken. Effectivity of treatments was assessed by addition of phytobiotic (control, 015% zinc bacitracin, 2.5% TE, 2.0% GE, 2.5% TGE which were arranged Completely Randomized Design with 4 replications. The variables measured were pancreatic enzyme activity(amylase enzyme activity, protease enzyme activity  and lipase enzyme activity.The results showed that enzyme protein activity content of 2.5% TE supplementation is also high at 82.02 U/ml, then supplemented 2.5% TGE, 2.0% GE, negative control and positive control respectively 75.98 ; 72.02; 68.74; and 66.57 U/ml. The lipase enzyme activity whereas the negative control and a positive control differ significantly higher (P<0.05 to treatment with the addition of 2.5% TE, 2.0% GE and 2.5% TGE phytobiotic. The research concluded that the incorporation of 2.5% TE, 2% GE and combined 2.5% TGE as feed additive enhanced pancreatic enzyme activity.

[The restoration of the enzyme activity of chernozem soil after gamma-irradiation].

Science.gov (United States)

Denisova, T V; Kazeev, K Sh

2006-01-01

The Influence of gamma-radiation by dozes 1, 5, 10 and 20 kGy on enzyme activity of ordinary chemozem were studied. Dynamics of the restoration of the enzyme activity after the influence of gamma-radiation in model experiments in 3, 30, 90 and 180 days was investigated. The doze 1 kGy did no statistically significant influence on the investigated enzymes. Dehydrogenase is more radiosensitive enzyme than catalase. Values of the saccharase activity differed a significant variation so in most cases it has not been registered statistically significant changes. In 90-180 days of the incubation enzymes activity was restored up to control values. Dehydrogenase activity in 180 days in variants with dozes 10 and 20 kGy was restored up to a level of the control, over variants with dozes 1 and 5 kGy--is higher than the control over 78% and 23% accordingly. Saccharase activity in 180 days after the influence of gamma-radiation with a doze 20 kGy was on 61% lower than the control.

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

Directory of Open Access Journals (Sweden)

Daryl L Moorhead

2013-08-01

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

An evaluation on elastase enzyme activity in gingival crevicular fluid in periodontitis

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Qujeq D

2003-08-01

Full Text Available Statement of Problem: Changes in protein levels, host calls enzymes and inflammatory mediators in gingival"ncrevicular Fluid (GCF are considered as diagnostic indicators of Periodontitis."nPurpose: he aim of the present study was to measure the elastase enzyme activity in gingival crevicular Fluid"namong patients with periodontitis."nMaterial and Methods: In this study, 52 periodontitis patients (experimental group and 51 healthy subjects"nwithout any gingival inflammatio (control group were participated. Subjects of the periodontitis group"nshowed pockets of 4-5 mm depth without gingival enlargement and recession or pockets of 1-2 mm depth"nwith gingival recession. For enzyme activity measurement, lOOu,! of gingival fluid of each sample was mixed"nwith lOOu! of enzyme substrate on the tube. The mixture was incubated at 34°c for lh with a buffer solution"nof 1ml volume and absorbance was read at 410nm with spectrophotometer. The enzyme activity differences"nbetween two groups were analyzed by student t test."nResults: The elastase enzyme activity in gingival crevicular fluid in subjects with periodontium destruction"nand control subjects was 153±11.3 and 52.7±10.4 enzyme unit in ml per minute, respectively. The difference"nbetween groups was statistically significant (PO.05."nConclusion: Based on the findings of this study, the measurement of elastae enzyme activity could be a useful"nindication of tissue changes that may ultimately manifest clinically as periodontitis.

Establishing Ideal Conditions for Complete Denitrification by Pseudomonas Aureofaciens - An Update on Determining Isotopic Composition of Dissolved Nitrate Using Bacterial Denitrification and Laser Spectroscopy

International Nuclear Information System (INIS)

Yi, Amelia Lee Zhi; Heiling, Maria; Toloza, Arsenio; Heng, Lee K.

2017-01-01

This serves as update on research entitled “Determining isotopic composition of dissolved nitrate using bacterial denitrification and laser spectroscopy” first published in the Vol. 39, No. 1, July 2016 SWMCN Soils Newsletter. In this research, isotopic δ"1"5N and δ"1"8O composition of dissolved nitrates is measured by laser spectroscopy after reduction of nitrate to N_2O by Pseudomonas aureofaciens. Quantifying the isotopic composition of nitrates in aqueous samples allows for better identification of potential nitrate sources, which in turn assists in remediation of nitrate-contaminated water and design of future agricultural management practises. The overall objective of the project is to establish a technical guide in the form of a standard operating procedure outlining best practises for denitrification method.

Remote enzyme activation using gold coated magnetite as antennae for radio frequency fields

Science.gov (United States)

Collins, Christian B.; Ackerson, Christopher J.

2018-02-01

The emerging field of remote enzyme activation, or the ability to remotely turn thermophilic increase enzyme activity, could be a valuable tool for understanding cellular processes. Through exploitation of the temperature dependence of enzymatic processes and high thermal stability of thermophilic enzymes these experiments utilize nanoparticles as `antennae' that convert radiofrequency (RF) radiation into local heat, increasing activity of the enzymes without increasing the temperature of the surrounding bulk solution. To investigate this possible tool, thermolysin, a metalloprotease was covalently conjugated to 4nm gold coated magnetite particles via peptide bond formation with the protecting ligand shell. RF stimulated protease activity at 17.76 MHz in a solenoid shaped antenna, utilizing both electric and magnetic field interactions was investigated. On average 40 percent higher protease activity was observed in the radio frequency fields then when bulk heating the sample to the same temperature. This is attributed to electrophoretic motion of the nanoparticle enzyme conjugates and local regions of heat generated by the relaxation of the magnetite cores with the oscillating field. Radio frequency local heating of nanoparticles conjugated to enzymes as demonstrated could be useful in the activation of specific enzymes in complex cellular environments.

Performance of heterotrophic partial denitrification under feast-famine condition of electron donor: a case study using acetate as external carbon source.

Science.gov (United States)

Gong, Lingxiao; Huo, Mingxin; Yang, Qing; Li, Jun; Ma, Bin; Zhu, Rulong; Wang, Shuying; Peng, Yongzhen

2013-04-01

Recently, the combination of anammox and post heterotrophic partial denitrification (nitrate to nitrite) was increasingly popular to treat anammox effluent with excessive nitrate, whereas achieving nitrite accumulation stably was a major bottleneck for post-denitrification. This work focused on the performance of heterotrophic partial denitrification under acetate feast-famine condition. The results showed that readily biodegradable COD to nitrate (RBCOD/NO3(-)) ratio of 2.5 facilitated an ideal nitrite accumulation ratio (NAR) of 71.7% under complete nitrate reduction. When RBCOD/NO3(-) ratio was below 3.5, in terms of efficiency and nitrite accumulation, higher NAR obtained during exogenous denitrification identified that the external acetate depletion was the optimal ending point of denitrification, which could be indicated by pH accurately. The indication of pH realized NAR of 60% ideally under batch-flow mode with RBCOD/NO3(-) ratio of 2.7, which might promote the scale-up of partial denitrification. Furthemore, alkaline environment (pH 9.0-9.6) repressed N2O emission even during endogenous denitrification. Copyright © 2012 Elsevier Ltd. All rights reserved.

Ligninolytic enzyme activities in mycelium of some wild and ...

African Journals Online (AJOL)

Lignin is probably one of the most recalcitrant compounds synthesized by plants. This compound is degraded by few microorganisms. White-rot fungi have been extensively studied due to its powerful ligninolytic enzymes. In this study, ligninolytic enzyme activities of different fungal species (six commercial and 13 wild) were ...

Alternative solutions for the bio-denitrification of landfill leachates using pine bark and compost

International Nuclear Information System (INIS)

Trois, Cristina; Pisano, Giulia; Oxarango, Laurent

2010-01-01

Nitrified leachate may still require an additional bio-denitrification step, which occurs with the addition of often-expensive chemicals as carbon source. This study explores the applicability of low-cost carbon sources such as garden refuse compost and pine bark for the denitrification of high strength landfill leachates. The overall objective is to assess efficiency, kinetics and performance of the substrates in the removal of high nitrate concentrations. Garden refuse and pine bark are currently disposed of in general waste landfills in South Africa, separated from the main waste stream. A secondary objective is to assess the feasibility of re-using green waste as by-product of an integrated waste management system. Denitrification processes in fixed bed reactors were simulated at laboratory scale using anaerobic batch tests and leaching columns packed with immature compost and pine bark. Biologically treated leachate from a Sequencing Batch Reactor (SBR) with nitrate concentrations of 350, 700 and 1100 mgN/l were used for the trials. Preliminary results suggest that, passed the acclimatization step (40 days for both substrates), full denitrification is achieved in 10-20 days for the pine bark and 30-40 days for the compost.

Activity of selected hydrolytic enzymes in Allium sativum L. anthers.

Science.gov (United States)

Winiarczyk, Krystyna; Gębura, Joanna

2016-05-01

The aim of the study was to determine enzymatic activity in sterile Allium sativum anthers in the final stages of male gametophyte development (the stages of tetrads and free microspores). The analysed enzymes were shown to occur in the form of numerous isoforms. In the tetrad stage, esterase activity was predominant, which was manifested by the greater number of isoforms of the enzyme. In turn, in the microspore stage, higher numbers of isoforms of acid phosphatases and proteases were detected. The development of sterile pollen grains in garlic is associated with a high level of protease and acid phosphatase activity and lower level of esterase activities in the anther locule. Probably this is the first description of the enzymes activity (ACPH, EST, PRO) in the consecutives stages of cell wall formation which is considered to be one of the causes of male sterility in flowering plant. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

Denitrification potential and its relation to organic carbon quality in three coastal wetland soils

Energy Technology Data Exchange (ETDEWEB)

Dodla, Syam K. [School of Plant, Environmental and Soil Sciences, Louisiana State Univ. Agricultural Center, Baton Rouge, LA 70803 (United States); Wang, Jim J. [School of Plant, Environmental and Soil Sciences, Louisiana State Univ. Agricultural Center, Baton Rouge, LA 70803 (United States)], E-mail: jjwang@agctr.lsu.edu; DeLaune, Ron D. [Wetland Biogeochemistry Institute, School of the Coast and Environment, Louisiana State University, Baton Rouge, LA 70803 (United States); Cook, Robert L. [Chemistry Department, Louisiana State University, Baton Rouge, LA 70803 (United States)

2008-12-15

Capacity of a wetland to remove nitrate through denitrification is controlled by its physico-chemical and biological characteristics. Understanding these characteristics will help better to guide beneficial use of wetlands in processing nitrate. This study was conducted to determine the relationship between soil organic carbon (SOC) quality and denitrification rate in Louisiana coastal wetlands. Composite soil samples of different depths were collected from three different wetlands along a salinity gradient, namely, bottomland forest swamp (FS), freshwater marsh (FM), and saline marsh (SM) located in the Barataria Basin estuary. Potential denitrification rate (PDR) was measured by acetylene inhibition method and distribution of carbon (C) moieties in organic C was determined by {sup 13}C solid-state NMR. Of the three wetlands, the FM soil profile exhibited the highest PDR on both unit weight and unit volume basis as compared to FS and SM. The FM also tended to yield higher amount of N{sub 2}O as compared to the FS and SM especially at earlier stages of denitrification, suggesting incomplete reduction of NO{sub 3}{sup -} at FM and potential for emission of N{sub 2}O. Saline marsh soil profile had the lowest PDR on the unit volume basis. Increasing incubation concentration from 2 to 10 mg NO{sub 3}{sup -}-N L{sup -1} increased PDR by 2 to 6 fold with the highest increase in the top horizons of FS and SM soils. Regression analysis showed that across these three wetland systems, organic C has significant effect in regulating PDR. Of the compositional C moieties, polysaccharides positively influenced denitrification rate whereas phenolics (likely phenolic adehydes and ketonics) negatively affected denitrification rate in these wetland soils. These results could have significant implication in integrated assessment and management of wetlands for treating nutrient-rich biosolids and wastewaters, non-point source agricultural runoff, and nitrate found in the diverted

Optimization of Enzyme Co-Immobilization with Sodium Alginate and Glutaraldehyde-Activated Chitosan Beads.

Science.gov (United States)

Gür, Sinem Diken; İdil, Neslihan; Aksöz, Nilüfer

2018-02-01

In this study, two different materials-alginate and glutaraldehyde-activated chitosan beads-were used for the co-immobilization of α-amylase, protease, and pectinase. Firstly, optimization of multienzyme immobilization with Na alginate beads was carried out. Optimum Na alginate and CaCl 2 concentration were found to be 2.5% and 0.1 M, respectively, and optimal enzyme loading ratio was determined as 2:1:0.02 for pectinase, protease, and α-amylase, respectively. Next, the immobilization of multiple enzymes on glutaraldehyde-activated chitosan beads was optimized (3% chitosan concentration, 0.25% glutaraldehyde with 3 h of activation and 3 h of coupling time). While co-immobilization was successfully performed with both materials, the specific activities of enzymes were found to be higher for the enzymes co-immobilized with glutaraldehyde-activated chitosan beads. In this process, glutaraldehyde was acting as a spacer arm. SEM and FTIR were used for the characterization of activated chitosan beads. Moreover, pectinase and α-amylase enzymes immobilized with chitosan beads were also found to have higher activity than their free forms. Three different enzymes were co-immobilized with these two materials for the first time in this study.

Chironomus plumosus larvae increase fluxes of denitrification products and diversity of nitrate-reducing bacteria in freshwater sediment

DEFF Research Database (Denmark)

Poulsen, Morten; W. V. Kofoed, Michael; H. Larsen, Lone

2014-01-01

, respectively, which was mostly due to stimulation of sedimentary denitrification; incomplete denitrification in the guts accounted for up to 20% of the N2O efflux. Phylotype richness of the nitrate reductase gene narG was significantly higher in sediment with than without larvae. In the gut, 47 narG phylotypes...... were found expressed, which may contribute to higher phylotype richness in colonized sediment. In contrast, phylotype richness of the nitrous oxide reductase gene nosZ was unaffected by the presence of larvae and very few nosZ phylotypes were expressed in the gut. Gene abundance of neither narG, nor...... nosZ wasdifferent in sediments with and without larvae. Hence, C. plumosus increases activity and diversity, but not overall abundance of nitrate-reducing bacteria, probably by providing additional ecological niches in its burrow and gut....

Using Pure Cultures to Define the Site Preference of Nitrous Oxide Produced by Microbial Nitrification and Denitrification

Science.gov (United States)

Sutka, R. L.; Breznak, J. A.; Ostrom, N. E.; Ostrom, P. H.; Gandhi, H.

2004-12-01

Defining the site preference of nitrous oxide (N2O) produced in pure culture studies is crucial to interpreting field data. We have previously demonstrated that the intramolecular distribution of nitrogen isotopes (isotopomers) can be used to differentiate N2O produced by nitrifier denitrification and nitrification in cultures of Nitrosomonas europaea. Here, we have expanded on our initial results and evaluated the isotopomeric composition of N2O produced during nitrification and nitrifier denitrification with cultures of Nitrosospira multiformis. In addition, we have analyzed N2O produced during methanotrophic nitrification, denitrification, and fungal denitrification. To evaluate N2O production during nitrification and nitrifier denitrification, we compared the site preference of N2O formed as a result of nitrite reduction and hydroxylamine oxidation with Nitrosomonas europaea and Nitrosospira multiformis. The average site preference of N2O produced by hydroxylamine oxidation was similar for Nitrosomonas europaea (33.0 ± 3.5 ‰ ) and Nitrosospira multiformis (33.1 ± 4.2 ‰ ). Nitrous oxide produced by nitrifier-denitrification by Nitrosomonas europaea and Nitrosospira multiformis had a similar site preference of - 1.4 ± 4.4 ‰ and - 1.1 ± 2.6 ‰ respectively. The results indicate that it is possible to differentiate between N2O produced by nitrite reduction and hydroxylamine oxidation by ammonia oxidizing bacteria. Methanotrophic nitrification was evaluated by analyzing the N2O produced during hydroxylamine oxidation in concentrated cell suspensions of two methane oxidizing bacteria. The site preference of N2O produced by the two methane oxidizers, Methylococcus capsulatus Bath and Methylosinus trichosporium was 31.8 ± 4.7 ‰ and 33.0 ± 4.5 ‰ respectively. The results indicate that a site preference of 33 ‰ is applicable for nitrification regardless of whether a methane oxidizer or ammonia oxidizer is involved in the reaction. To determine the site

Revealing hidden effect of earthworm on C distribution and enzyme activity

Science.gov (United States)

Razavi, Bahar S.; Hoang, Duyen; Kuzyakov, Yakov

2017-04-01

Despite its importance for terrestrial nutrient and carbon cycling, the spatial organization and localization of microbial activity in soil and in biopores is poorly understood. We hypothesized that biopores created by earthworm play a critical role in reducing the gap of SOM input and microbial activities between topsoil and subsoil. Accordingly, Carbon (C) allocation by earthworms was related to enzyme distribution along soil profile. For the first time we visualized spatial distribution of enzyme activities (β-glucosidase, chitinase and acid phosphatase) and C allocation (by 14C imaging) in earthworm biopores in topsoil and subsoil. Soil zymography (an in situ method for the analysis of the two-dimensional distribution of enzyme activity in soil) was accompanied with 14C imaging (a method that enables to trace distribution of litter and C in soil profile) to visualize change of enzyme activities along with SOM incorporation by earthworms from topsoil to subsoil. Experiment was set up acquiring rhizoboxes (9×1×50 cm) filled up with fresh soil and 3 earthworms (L. terrestris), which were then layered with 14C-labeled plant-litter of 0.3 MBq on the soil surface. 14C imaging and zymography have been carried out after one month. Activities of all enzymes regardless of their nutrient involvement (C, N, P) were higher in the biopores than in bulk soil, but the differences were larger in topsoil compared to subsoil. Among three enzymes, Phosphatase activity was 4-times higher in the biopore than in the bulk soil. Phosphatase activity was closely associated with edge of burrows and correlate positively with 14C activity. These results emphasized especial contribution of hotspheres such as biopores to C allocation in subsoil - which is limited in C input and nutrients - and in stimulation of microbial and enzymatic activity by input of organic residues, e.g. by earthworms. In conclusion, biopore increased enzymatic mobilization of nutrients (e.g. P) inducing allocation

Molecular dynamics explorations of active site structure in designed and evolved enzymes.

Science.gov (United States)

Osuna, Sílvia; Jiménez-Osés, Gonzalo; Noey, Elizabeth L; Houk, K N

2015-04-21

This Account describes the use of molecular dynamics (MD) simulations to reveal how mutations alter the structure and organization of enzyme active sites. As proposed by Pauling about 70 years ago and elaborated by many others since then, biocatalysis is efficient when functional groups in the active site of an enzyme are in optimal positions for transition state stabilization. Changes in mechanism and covalent interactions are often critical parts of enzyme catalysis. We describe our explorations of the dynamical preorganization of active sites using MD, studying the fluctuations between active and inactive conformations normally concealed to static crystallography. MD shows how the various arrangements of active site residues influence the free energy of the transition state and relates the populations of the catalytic conformational ensemble to the enzyme activity. This Account is organized around three case studies from our laboratory. We first describe the importance of dynamics in evaluating a series of computationally designed and experimentally evolved enzymes for the Kemp elimination, a popular subject in the enzyme design field. We find that the dynamics of the active site is influenced not only by the original sequence design and subsequent mutations but also by the nature of the ligand present in the active site. In the second example, we show how microsecond MD has been used to uncover the role of remote mutations in the active site dynamics and catalysis of a transesterase, LovD. This enzyme was evolved by Tang at UCLA and Codexis, Inc., and is a useful commercial catalyst for the production of the drug simvastatin. X-ray analysis of inactive and active mutants did not reveal differences in the active sites, but relatively long time scale MD in solution showed that the active site of the wild-type enzyme preorganizes only upon binding of the acyl carrier protein (ACP) that delivers the natural acyl group to the active site. In the absence of bound ACP

Denitrification in the karstic Floridan Aquifer

Science.gov (United States)

Fork, M.; Albertin, A. R.; Heffernan, J. B.; Katz, B. G.; Cohen, M. J.

2010-12-01

Nitrate concentrations in the karstic Floridan Aquifer have increased dramatically over the past 50 years, owing to agricultural intensification and urbanization. Due to low concentrations of organic matter and moderately oxic conditions in the Floridan Aquifer, groundwater denitrification has been assumed to be negligible. In this study, we evaluate that assumption using both existing and new data describing dissolved gases (Ne, N2, O2, Ar) and NO3- concentration and isotopic composition (δ18O- and δ15N-NO3) in the aquifer’s artesian springs. For new data, we collected samples from 33 spring vents representing a gradient of both DO and NO3- concentrations in northern Florida and used Membrane Inlet Mass Spectrometry (MIMS) to directly measure dissolved N2 and Ar. We modeled the physical processes (recharge temperature, dissolution of excess air) driving super-saturation of N2 gas using Ne and Ar where data describing Ne were available. Ar concentrations were correlated closely with recharge temperature, which ranged from 15.7 - 22.2°C, while Ne was closely correlated with excess air, which ranged from 1.05 to 2.66 mg L-1 and averaged 1.83 mg L-1. Estimates of physical mechanisms allowed calculation of expected N2 concentrations that were compared to observed N2 concentrations. Where Ne data were unavailable, we assumed excess air equal to the empirical average. Overall, observed N2 exceeded expectations based on physical processes in 33 of 47 cases; average excess N2 was 0.48 mg L-1 across all sites. In addition, excess N2 was negatively correlated with DO (r2 = 0.46); springs with low DO (Aquifer. Low DOC concentrations indicate that alternative electron donors may fuel nitrate reduction. Scaling to regional estimates of N2 production based on springs discharge and DO concentrations indicates that subsurface denitrification may account for some of the imbalance in springshed nutrient budgets. In addition, we conclude that use of δ15N-NO3- to diagnose

Nitrous oxide production by nitrification and denitrification in the Eastern Tropical South Pacific oxygen minimum zone

Science.gov (United States)

Ji, Qixing; Babbin, Andrew R.; Jayakumar, Amal; Oleynik, Sergey; Ward, Bess B.

2015-12-01

The Eastern Tropical South Pacific oxygen minimum zone (ETSP-OMZ) is a site of intense nitrous oxide (N2O) flux to the atmosphere. This flux results from production of N2O by nitrification and denitrification, but the contribution of the two processes is unknown. The rates of these pathways and their distributions were measured directly using 15N tracers. The highest N2O production rates occurred at the depth of peak N2O concentrations at the oxic-anoxic interface above the oxygen deficient zone (ODZ) because slightly oxygenated waters allowed (1) N2O production from both nitrification and denitrification and (2) higher nitrous oxide production yields from nitrification. Within the ODZ proper (i.e., anoxia), the only source of N2O was denitrification (i.e., nitrite and nitrate reduction), the rates of which were reflected in the abundance of nirS genes (encoding nitrite reductase). Overall, denitrification was the dominant pathway contributing the N2O production in the ETSP-OMZ.

Spatial distribution of enzyme activities along the root and in the rhizosphere of different plants

Science.gov (United States)

Razavi, Bahar S.; Zarebanadkouki, Mohsen; Blagodatskaya, Evgenia; Kuzyakov, Yakov

2015-04-01

Extracellular enzymes are important for decomposition of many biological macromolecules abundant in soil such as cellulose, hemicelluloses and proteins. Activities of enzymes produced by both plant roots and microbes are the primary biological drivers of organic matter decomposition and nutrient cycling. So far acquisition of in situ data about local activity of different enzymes in soil has been challenged. That is why there is an urgent need in spatially explicit methods such as 2-D zymography to determine the variation of enzymes along the roots in different plants. Here, we developed further the zymography technique in order to quantitatively visualize the enzyme activities (Spohn and Kuzyakov, 2013), with a better spatial resolution We grew Maize (Zea mays L.) and Lentil (Lens culinaris) in rhizoboxes under optimum conditions for 21 days to study spatial distribution of enzyme activity in soil and along roots. We visualized the 2D distribution of the activity of three enzymes:β-glucosidase, leucine amino peptidase and phosphatase, using fluorogenically labelled substrates. Spatial resolution of fluorescent images was improved by direct application of a substrate saturated membrane to the soil-root system. The newly-developed direct zymography shows different pattern of spatial distribution of enzyme activity along roots and soil of different plants. We observed a uniform distribution of enzyme activities along the root system of Lentil. However, root system of Maize demonstrated inhomogeneity of enzyme activities. The apical part of an individual root (root tip) in maize showed the highest activity. The activity of all enzymes was the highest at vicinity of the roots and it decreased towards the bulk soil. Spatial patterns of enzyme activities as a function of distance from the root surface were enzyme specific, with highest extension for phosphatase. We conclude that improved zymography is promising in situ technique to analyze, visualize and quantify

Seasonal effects of the zebra mussel (Dreissena polymorpha) on sediment denitrification rates in Pool 8 of the Upper Mississippi River

Science.gov (United States)

Bruesewitz, Denise A.; Tank, Jennifer L.; Bernot, Melody J.; Richardson, William B.; Strauss, Eric A.

2006-01-01

Zebra mussels (Dreissena polymorpha) have altered the structure of invaded ecosystems and exhibit characteristics that suggest they may influence ecosystem processes such as nitrogen (N) cycling. We measured denitrification rates seasonally on sediments underlying zebra mussel beds collected from the impounded zone of Navigation Pool 8 of the Upper Mississippi River. Denitrification assays were amended with nutrients to characterize variation in nutrient limitation of denitrification in the presence or absence of zebra mussels. Denitrification rates at zebra mussel sites were high relative to sites without zebra mussels in February 2004 (repeated measures analysis of variance (RM ANOVA), p = 0.005), potentially because of high NO3-N variability from nitrification of high NH4+ zebra mussel waste. Denitrification rates were highest in June 2003 (RM ANOVA, p 3-N concentrations during the study (linear regression, R2 = 0.72, p p ≤ 0.01). Examining how zebra mussels influence denitrification rates will aid in developing a more complete understanding of the impact of zebra mussels and more effective management strategies of eutrophic waters.

A Tire-Sulfur Hybrid Adsorption Denitrification (T-SHAD) process for decentralized wastewater treatment.

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Krayzelova, Lucie; Lynn, Thomas J; Banihani, Qais; Bartacek, Jan; Jenicek, Pavel; Ergas, Sarina J

2014-09-15

Nitrogen discharges from decentralized wastewater treatment (DWT) systems contribute to surface and groundwater contamination. However, the high variability in loading rates, long idle periods and lack of regular maintenance presents a challenge for biological nitrogen removal in DWT. A Tire-Sulfur Hybrid Adsorption Denitrification (T-SHAD) process was developed that combines nitrate (NO3(-)) adsorption to scrap tire chips with sulfur-oxidizing denitrification. This allows the tire chips to adsorb NO3(-) when the influent loading exceeds the denitrification capacity of the biofilm and release it when NO3(-) loading rates are low (e.g. at night). Three waste products, scrap tire chips, elemental sulfur pellets and crushed oyster shells, were used as a medium in adsorption, leaching, microcosm and up-flow packed bed bioreactor studies of NO3(-) removal from synthetic nitrified DWT wastewater. Adsorption isotherms showed that scrap tire chips have an adsorption capacity of 0.66 g NO3(-)-N kg(-1) of scrap tires. Leaching and microcosm studies showed that scrap tires leach bioavailable organic carbon that can support mixotrophic metabolism, resulting in lower effluent SO4(2-) concentrations than sulfur oxidizing denitrification alone. In column studies, the T-SHAD process achieved high NO3(-)-N removal efficiencies under steady state (90%), variable flow (89%) and variable concentration (94%) conditions. Copyright © 2014 Elsevier Ltd. All rights reserved.

Influence of long-term fertilization on soil enzyme activities

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Alina Dora SAMUEL

2009-05-01

Full Text Available Soil enzyme activities (actual and potential dehydrogenase, catalase, acid and alkaline phosphatase were determined in the 0–10, 10–20, and 20–30 cm layers of a brown luvic soil submitted to a complex fertilization experiment with different types of green manure. It was found that each activity decreased with increasing sampling depth. It should be emphasized that greenmanuring of maize led to a significant increase in each of the five enzymatic activities determined. The enzymatic indicators of soil quality calculated from the values of enzymatic activities showed the order: lupinus + rape + oat > lupinus > vetch + oat + ryegrass > lupinus + oat + vetch > unfertilized plot. This order means that by determination of enzymatic activities valuable information can be obtained regarding fertility status of soils. There were significant correlations of soil enzyme activities with chemical properties.

15N kinetic analysis of N2O production by Nitrosomonas europaea: an examination of nitrifier denitrification

International Nuclear Information System (INIS)

Poth, M.; Focht, D.D.

1985-01-01

A series of 15 N isotope tracer experiments showed that Nitrosomonas europaea produces nitrous oxide only under oxygen-limiting conditions and that the labeled N from nitrite, but not nitrate, is incorporated into nitrous oxide, indicating the presence of the denitrifying enzyme nitrite reductase. A kinetic analysis of the m/z 44, 45, and 46 nitrous oxide produced by washed cell suspensions of N. europaea when incubated with 4 mM ammonium (99% 14 N) and 0.4 mM nitrite (99% 15 N) was performed. No labeled nitirte was reduced to ammonium. All labeled material added was accounted for as either nitrite or nitrous oxide. The hypothesis that nitrous oxide is produced directly from nitrification was rejected since (i) it does not allow for the large amounts of double-labeled (m/z 46) nitrous oxide observed; (ii) the observed patterns of m/z 44, 45, 46 nitrous oxide were completely consistent with a kinetic analysis based on denitrification as the sole mechanism of nitrous oxide production but not with a kinetic analysis based on both mechanisms; (iii) the asymptotic ratio of m/z 45 to m/z 46 nitrous oxide was consistent with denitrification kinetics but inconsistent with nitrification kinetics, which predicted no limit to m/z 45 production. It is concluded that N. europaea is a denitrifier which, under conditions of oxygen stress, uses nitrite as a terminal electron acceptor and produces nitrous oxide

A metal-based inhibitor of NEDD8-activating enzyme.

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Hai-Jing Zhong

Full Text Available A cyclometallated rhodium(III complex [Rh(ppy(2(dppz](+ (1 (where ppy=2-phenylpyridine and dppz=dipyrido[3,2-a:2',3'-c]phenazine dipyridophenazine has been prepared and identified as an inhibitor of NEDD8-activating enzyme (NAE. The complex inhibited NAE activity in cell-free and cell-based assays, and suppressed the CRL-regulated substrate degradation and NF-κB activation in human cancer cells with potency comparable to known NAE inhibitor MLN4924. Molecular modeling analysis suggested that the overall binding mode of 1 within the binding pocket of the APPBP1/UBA3 heterodimer resembled that for MLN4924. Complex 1 is the first metal complex reported to suppress the NEDDylation pathway via inhibition of the NEDD8-activating enzyme.

Enzyme activity screening of thermophilic bacteria isolated from Dusun Tua Hot Spring, Malaysia

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Msarah, Marwan; Ibrahim, Izyanti; Aqma, Wan Syaidatul

2018-04-01

Thermophilic bacteria have biotechnological importance due to the availability of unique enzymes which are stable in extreme circumstances. The aim of this study includes to isolate thermophilic bacteria from hot spring and screen for important enzyme activities. Water samples from the Dusun Tua Hot Spring were collected and the physiochemical characterisation of water was measured. Eight thermophilic bacteria were isolated and determined to have at least three strong enzyme activity including protease, lipase, amylase, cellulase, pectinase and xylanase. The results showed that HuluC2 displayed all the enzyme activities and can be further studied.

Understanding drivers of peatland extracellular enzyme activity in the PEATcosm experiment: mixed evidence for enzymic latch hypothesis

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Karl J. Romanowicz; Evan S. Kane; Lynette R. Potvin; Aleta L. Daniels; Randy Kolka; Erik A. Lilleskov

2015-01-01

Aims. Our objective was to assess the impacts of water table position and plant functional groups on peatland extracellular enzyme activity (EEA) framed within the context of the enzymic latch hypothesis. Methods. We utilized a full factorial experiment with 2 water table (WT) treatments (high and low) and 3 plant functional...

Subcellular distribution of histone-degrading enzyme activities from rat liver

International Nuclear Information System (INIS)

Heinrich, P.C.; Raydt, G.; Puschendorf, B.; Jusic, M.

1976-01-01

Chromatin prepared from liver tissue contains a histone-degrading enzyme activity with a pH optimum of 7.5-8.0, whereas chromatin isolated from purified nuclei is devoid of it. The histone-degrading enzyme activity was assayed with radioactively labelled total histones from Ehrlich ascites tumor cells. Among the different subcellular fractions assayed, only lysosomes and mitochondria exhibited histone-degrading enzymes. A pH optimum around 4.0-5.0 was found for the lysosomal fraction, whereas 7.5-8.0 has been found for mitochondria. Binding studies of frozen and thawed lysosomes or mitochondria to proteinase-free chromatin demonstrate that the proteinase associated with chromatin isolated from frozen tissue originates from damaged mitochondria. The protein degradation patterns obtained after acrylamide gel electrophoresis are similar for the chromatin-associated and the mitochondrial proteinase and different from that obtained after incubation with lysosomes. The chromatin-associated proteinase as well as the mitochondrial proteinase are strongly inhibited by 1.0 mM phenylmethanesulfonyl fluoride. Weak inhibition is found for lysosomal proteinases at pH 5. Kallikrein-trypsin inhibitor, however, inhibits lysosomal proteinase activity and has no effect on either chromatin-associated or mitochondrial proteinases. The higher template activity of chromatin isolated from a total homogenate compared to chromatin prepared from nuclei may be due to the presence of this histone-degrading enzyme activity. (orig.) [de

Gaseous environment of plants and activity of enzymes of carbohydrate catabolism

International Nuclear Information System (INIS)

Ivanov, B.F.; Zemlyanukhin, A.A.; Igamberdiev, A.U.; Salam, A.M.M.

1989-01-01

The authors investigated the action of hypoxia and high CO 2 concentration in the atmosphere on activity of phosphofructokinase, aldolase, glucose phosphate isomerase, glucose-6-phosphate dehydrogenase, lactate dehydrogenase, alcohol dehydrogenase, and isocitrate lyase in pea seedlings (Pisum sativum L.), corn scutella (Zea mays L.), and hemp cotyledons (Cannabis sativa L.). The first 4-12h of hypoxia witnessed suppression of enzymes of the initial stages of glycolysis (glucose-6-phosphate isomerase, phosphofructokinase)and activation of enzymes of its final stages (alcohol dehydrogenase and lactate dehydrogenase) and enzymes linking glycolysis and the pentose phosphate pathway (aldolase and glucose-6-phosphate dehydrogenase). An excess of CO 2 in the environment accelerated and amplified this effect. At the end of a 24-h period of anaerobic incubation, deviations of enzyme activity from the control were leveled in both gaseous environments. An exception was observed in the case of phosphofructokinase, whose activity increased markedly at this time in plants exposed to CO 2 . Changes in activity of the enzymes were coupled with changes in their kinetic parameters (apparent K m and V max values). The activity of isocitrate lyase was suppressed in both variants of hypoxic gaseous environments, a finding that does not agree with the hypothesis as to participation of the glyoxylate cycle in the metabolic response of plants to oxygen stress. Thus, temporary inhibition of the system of glycolysis and activation of the pentose phosphate pathway constituted the initial response of the plants to O 2 stress, and CO 2 intensified this metabolic response

Alternative solutions for the bio-denitrification of landfill leachates using pine bark and compost.

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Trois, Cristina; Pisano, Giulia; Oxarango, Laurent

2010-06-15

Nitrified leachate may still require an additional bio-denitrification step, which occurs with the addition of often-expensive chemicals as carbon source. This study explores the applicability of low-cost carbon sources such as garden refuse compost and pine bark for the denitrification of high strength landfill leachates. The overall objective is to assess efficiency, kinetics and performance of the substrates in the removal of high nitrate concentrations. Garden refuse and pine bark are currently disposed of in general waste landfills in South Africa, separated from the main waste stream. A secondary objective is to assess the feasibility of re-using green waste as by-product of an integrated waste management system. Denitrification processes in fixed bed reactors were simulated at laboratory scale using anaerobic batch tests and leaching columns packed with immature compost and pine bark. Biologically treated leachate from a Sequencing Batch Reactor (SBR) with nitrate concentrations of 350, 700 and 1100 mgN/l were used for the trials. Preliminary results suggest that, passed the acclimatization step (40 days for both substrates), full denitrification is achieved in 10-20 days for the pine bark and 30-40 days for the compost. Copyright 2010 Elsevier B.V. All rights reserved.

Ultrasound assisted intensification of enzyme activity and its properties: a mini-review.

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Nadar, Shamraja S; Rathod, Virendra K

2017-08-22

Over the last decade, ultrasound technique has emerged as the potential technology which shows large applications in food and biotechnology processes. Earlier, ultrasound has been employed as a method of enzyme inactivation but recently, it has been found that ultrasound does not inactivate all enzymes, particularly, under mild conditions. It has been shown that the use of ultrasonic treatment at appropriate frequencies and intensity levels can lead to enhanced enzyme activity due to favourable conformational changes in protein molecules without altering its structural integrity. The present review article gives an overview of influence of ultrasound irradiation parameters (intensity, duty cycle and frequency) and enzyme related factors (enzyme concentration, temperature and pH) on the catalytic activity of enzyme during ultrasound treatment. Also, it includes the effect of ultrasound on thermal kinetic parameters and Michaelis-Menten kinetic parameters (k m and V max ) of enzymes. Further, in this review, the physical and chemical effects of ultrasound on enzyme have been correlated with thermodynamic parameters (enthalpy and entropy). Various techniques used for investigating the conformation changes in enzyme after sonication have been highlighted. At the end, different techniques of immobilization for ultrasound treated enzyme have been summarized.

Sulfur-oxidizing autotrophic and mixotrophic denitrification processes for drinking water treatment: elimination of excess sulfate production and alkalinity requirement.

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Sahinkaya, Erkan; Dursun, Nesrin

2012-09-01

This study evaluated the elimination of alkalinity need and excess sulfate generation of sulfur-based autotrophic denitrification process by stimulating simultaneous autotrophic and heterotrophic (mixotrophic) denitrification process in a column bioreactor by methanol supplementation. Also, denitrification performances of sulfur-based autotrophic and mixotrophic processes were compared. In autotrophic process, acidity produced by denitrifying sulfur-oxidizing bacteria was neutralized by the external NaHCO(3) supplementation. After stimulating mixotrophic denitrification process, the alkalinity need of the autotrophic process was satisfied by the alkalinity produced by heterotrophic denitrifiers. Decreasing and lastly eliminating the external alkalinity supplementation did not adversely affect the process performance. Complete denitrification of 75 mg L(-1) NO(3)-N under mixotrophic conditions at 4 h hydraulic retention time was achieved without external alkalinity supplementation and with effluent sulfate concentration lower than the drinking water guideline value of 250 mg L(-1). The denitrification rate of mixotrophic process (0.45 g NO(3)-N L(-1) d(-1)) was higher than that of autotrophic one (0.3 g NO(3)-N L(-1) d(-1)). Batch studies showed that the sulfur-based autotrophic nitrate reduction rate increased with increasing initial nitrate concentration and transient accumulation of nitrite was observed. Copyright © 2012 Elsevier Ltd. All rights reserved.

River water infiltration enhances denitrification efficiency in riparian groundwater.

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Trauth, Nico; Musolff, Andreas; Knöller, Kay; Kaden, Ute S; Keller, Toralf; Werban, Ulrike; Fleckenstein, Jan H

2018-03-01

Nitrate contamination in ground- and surface water is a persistent problem in countries with intense agriculture. The transition zone between rivers and their riparian aquifers, where river water and groundwater interact, may play an important role in mediating nitrate exports, as it can facilitate intensive denitrification, which permanently removes nitrate from the aquatic system. However, the in-situ factors controlling riparian denitrification are not fully understood, as they are often strongly linked and their effects superimpose each other. In this study, we present the evaluation of hydrochemical and isotopic data from a 2-year sampling period of river water and groundwater in the riparian zone along a 3rd order river in Central Germany. Based on bi- and multivariate statistics (Spearman's rank correlation and partial least squares regression) we can show, that highest rates for oxygen consumption and denitrification in the riparian aquifer occur where the fraction of infiltrated river water and at the same time groundwater temperature, are high. River discharge and depth to groundwater are additional explanatory variables for those reaction rates, but of minor importance. Our data and analyses suggest that at locations in the riparian aquifer, which show significant river water infiltration, heterotrophic microbial reactions in the riparian zone may be fueled by bioavailable organic carbon derived from the river water. We conclude that interactions between rivers and riparian groundwater are likely to be a key control of nitrate removal and should be considered as a measure to mitigate high nitrate exports from agricultural catchments. Copyright © 2017 Elsevier Ltd. All rights reserved.

ATPase Activity Measurements by an Enzyme-Coupled Spectrophotometric Assay.

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Sehgal, Pankaj; Olesen, Claus; Møller, Jesper V

2016-01-01

Enzymatic coupled assays are usually based on the spectrophotometric registration of changes in NADH/NAD(+) or NADPH/NADP(+) absorption at 340 nm accompanying the oxidation/reduction of reactants that by dehydrogenases and other helper enzymes are linked to the activity of the enzymatic reaction under study. The present NADH-ATP-coupled assay for ATPase activity is a seemingly somewhat complicated procedure, but in practice adaptation to performance is easily acquired. It is a more safe and elegant method than colorimetric methods, but not suitable for handling large number of samples, and also presupposes that the activity of the helper enzymes is not severely affected by the chemical environment of the sample in which it is tested.

Interrogating the activities of conformational deformed enzyme by single-molecule fluorescence-magnetic tweezers microscopy

Science.gov (United States)

Guo, Qing; He, Yufan; Lu, H. Peter

2015-01-01

Characterizing the impact of fluctuating enzyme conformation on enzymatic activity is critical in understanding the structure–function relationship and enzymatic reaction dynamics. Different from studying enzyme conformations under a denaturing condition, it is highly informative to manipulate the conformation of an enzyme under an enzymatic reaction condition while monitoring the real-time enzymatic activity changes simultaneously. By perturbing conformation of horseradish peroxidase (HRP) molecules using our home-developed single-molecule total internal reflection magnetic tweezers, we successfully manipulated the enzymatic conformation and probed the enzymatic activity changes of HRP in a catalyzed H2O2–amplex red reaction. We also observed a significant tolerance of the enzyme activity to the enzyme conformational perturbation. Our results provide a further understanding of the relation between enzyme behavior and enzymatic conformational fluctuation, enzyme–substrate interactions, enzyme–substrate active complex formation, and protein folding–binding interactions. PMID:26512103

Heterogeneity of hydrolytic enzyme activities under drought: imaging and quantitative analysis

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Sanaullah, Muhammad; Razavi, Bahar S.; Kuzyakov, Yakov

2015-04-01

The zymography-based "snap-shoot" of enzyme activities in the rhizosphere is challenging to detect the in situ microbial response to global climate change. We developed in situ soil zymography and used it for identification and localization of hotspots of β-glucosidase activity in the rhizosphere of maize under drought stress (30% of field capacity). The zymographic signals were especially high at root tips and were much stronger for activity of β-glucosidase under drought as compared with optimal moisture (70% of field capacity). This distribution of enzyme activity was confirmed by fluorogenically labelled substrates applied directly to the root exudates. The activity of β-glucosidase in root exudates (produced by root and microorganism associated on the root surface), sampled within 1 hour after zymography was significantly higher by drought stressed plants as compared with optimal moisture. In contrast, the β-glucosidase activity in destructively sampled rhizosphere soil was lower under drought stress compared with optimal moisture. Furthermore, drought stress did not affected β-glucosidase activity in bulk soil, away from rhizosphere. Consequently, we conclude that higher release of mucilage by roots und drought stimulated β-glucosidase activity in the rhizosphere. Thus, the zymography revealed plant-mediated mechanisms accelerating β-glucosidase activity under drought at the root-soil interface. So, coupling of zymography and enzyme assays in the rhizosphere and non-rhizosphere soil enables precise mapping the changes in two-dimensional distribution of enzyme activities due to climate change within dynamic soil interfaces.

The Evolution of Indian and Pacific Ocean Denitrification and Nitrogen Dynamcs since the Miocene

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Ravelo, A. C.; Carney, C.; Rosenthal, Y.; Holbourn, A.; Kulhanek, D. K.

2017-12-01

The feedbacks between geochemical cycles and physical climate change are poorly understood; however, there has been tremendous progress in developing coupled models to help predict the direction and strength of these feedbacks. As such, there is a need for more data to validate and test these models. To this end, the nitrogen (N) cycle, and its links to the biological pump and to climate, is an active area of paleoceanographic research. Using N isotope records, Robinson et al. (2014) showed that pelagic denitrification in the Indian and Pacific Oceans intensified as climate cooled and subsurface ventilation decreased since the Pliocene. They pointed out that a more ventilated warm Pliocene contrasts with glacial-interglacial patterns wherein more ventilation occurs during cold phases, indicating that different mechanisms may occur at different timescales. Our objective is to better understand the nature of the feedbacks between the oceanic N cycle and climate by focusing on the large dynamic range of conditions that occurred during and since the Miocene. We used new cores drilled during IODP Expedition 363 to generate bulk sediment N isotope records at three western tropical Pacific sites (U1486, U1488, U1490) and one southeastern tropical Indian Ocean site (U1482). We find that the N isotope trends since the Pliocene are in agreement with previous studies showing increasing denitrification as climate cooled. In the Miocene, the Indian Ocean record shows no long-term N isotope trend whereas the Pacific Ocean records show a trend that is roughly coupled to changes in global climate suggesting that pelagic denitrification in the Pacific was strongly influenced by greater ventilation during global warmth. However, there are notable deviations from this coupling during several intervals in the Miocene, and there are site-to-site differences in trends. These deviations and differences can be explained by changes in tropical productivity (e.g., late Miocene biogenic

Adsorption and enzyme activity of asparaginase at lipid Langmuir and Langmuir-Blodgett films

International Nuclear Information System (INIS)

Rocha Junior, Carlos da; Caseli, Luciano

2017-01-01

In this present work, the surface activity of the enzyme asparaginase was investigated at the air-water interface, presenting surface activity in high ionic strengths. Asparaginase was incorporated in Langmuir monolayers of the phospholipid dipalmitoylphosphatidylcholine (DPPC), forming a mixed film, which was characterized with surface pressure-area isotherms, surface potential-area isotherms, polarization-modulated infrared reflection-absorption spectroscopy (PM-IRRAS), and Brewster angle microscopy (BAM). The adsorption of the enzyme at the air-water interface condensed the lipid monolayer and increased the film compressibility at high surface pressures. Amide bands in the PM-IRRAS spectra were identified, with the C−N and C =O dipole moments lying parallel to monolayer plane, revealing the structuring of the enzyme into α-helices and β-sheets. The floating monolayers were transferred to solid supports as Langmuir-Blodgett (LB) films and characterized with fluorescence spectroscopy and atomic force microscopy. Catalytic activities of the films were measured and compared to the homogenous medium. The enzyme accommodated in the LB films preserved more than 78% of the enzyme activity after 30 days, in contrast for the homogeneous medium, which preserved less than 13%. The method presented in this work not only allows for an enhanced catalytic activity, but also can help explain why certain film architectures exhibit better performance. - Highlights: • Biomembranes are mimicked with Langmuir monolayers. • Asparaginase is incorporated into the lipid monolayer. • Enzyme adsorption is confirmed with tensiometry and infrared spectroscopy. • Langmuir-Blodgett films of the enzyme present enzyme activity.

Adsorption and enzyme activity of asparaginase at lipid Langmuir and Langmuir-Blodgett films

Energy Technology Data Exchange (ETDEWEB)

Rocha Junior, Carlos da; Caseli, Luciano, E-mail: lcaseli@unifesp.br

2017-04-01

In this present work, the surface activity of the enzyme asparaginase was investigated at the air-water interface, presenting surface activity in high ionic strengths. Asparaginase was incorporated in Langmuir monolayers of the phospholipid dipalmitoylphosphatidylcholine (DPPC), forming a mixed film, which was characterized with surface pressure-area isotherms, surface potential-area isotherms, polarization-modulated infrared reflection-absorption spectroscopy (PM-IRRAS), and Brewster angle microscopy (BAM). The adsorption of the enzyme at the air-water interface condensed the lipid monolayer and increased the film compressibility at high surface pressures. Amide bands in the PM-IRRAS spectra were identified, with the C−N and C =O dipole moments lying parallel to monolayer plane, revealing the structuring of the enzyme into α-helices and β-sheets. The floating monolayers were transferred to solid supports as Langmuir-Blodgett (LB) films and characterized with fluorescence spectroscopy and atomic force microscopy. Catalytic activities of the films were measured and compared to the homogenous medium. The enzyme accommodated in the LB films preserved more than 78% of the enzyme activity after 30 days, in contrast for the homogeneous medium, which preserved less than 13%. The method presented in this work not only allows for an enhanced catalytic activity, but also can help explain why certain film architectures exhibit better performance. - Highlights: • Biomembranes are mimicked with Langmuir monolayers. • Asparaginase is incorporated into the lipid monolayer. • Enzyme adsorption is confirmed with tensiometry and infrared spectroscopy. • Langmuir-Blodgett films of the enzyme present enzyme activity.

Experimental Strategy to Discover Microbes with Gluten-degrading Enzyme Activities.

Science.gov (United States)

Helmerhorst, Eva J; Wei, Guoxian

2014-05-05

Gluten proteins contained in the cereals barley, rye and wheat cause an inflammatory disorder called celiac disease in genetically predisposed individuals. Certain immunogenic gluten domains are resistant to degradation by mammalian digestive enzymes. Enzymes with the ability to target such domains are potentially of clinical use. Of particular interest are gluten-degrading enzymes that would be naturally present in the human body, e.g. associated with resident microbial species. This manuscript describes a selective gluten agar approach and four enzyme activity assays, including a gliadin zymogram assay, designed for the selection and discovery of novel gluten-degrading microorganisms from human biological samples. Resident and harmless bacteria and/or their derived enzymes could potentially find novel applications in the treatment of celiac disease, in the form of a probiotic agent or as a dietary enzyme supplement.

Hydrolytic enzyme activity enhanced by Barium supplementation

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Camilo Muñoz

2016-10-01

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

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

Science.gov (United States)

Glass, Jennifer B.; Orphan, Victoria J.

2011-01-01

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

The role of C:N:P stoichiometry in affecting denitrification in sediments from agricultural surface and tile-water wetlands.

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Grebliunas, Brian D; Perry, William L

2016-01-01

Nutrient stoichiometry within a wetland is affected by the surrounding land use, and may play a significant role in the removal of nitrate (NO3-N). Tile-drained, agricultural watersheds experience high seasonal inputs of NO3-N, but low phosphorus (PO4-P) and dissolved organic carbon (DOC) loads relative to surface water dominated systems. This difference may present stoichiometric conditions that limit denitrification within receiving waterways. We investigated how C:N:P ratios affected denitrification rates of sediments from tile-drained mitigation wetlands incubated for: 0, 5, 10, and 20 days. We then tested whether denitrification rates of sediments from surface-water and tile-drained wetlands responded differently to C:N ratios of 2:1 versus 4:1. Ratios of C:N:P (P tile-drained wetland sediments. Carbon limitation of denitrification became evident at elevated NO3-N concentrations (20 mg L(-1)). Denitrification measured from tile water and surface water wetland sediments increased significantly (P < 0.05) at the 2:1 and 4:1 C:N treatments. The results from both experiments suggest wetland sediments provide a limiting pool of labile DOC to maintain prolonged NO3-N removal. Also, DOC limitation became more evident at elevated NO3-N concentrations (20 mg L(-1)). Irrespective of NO3-N concentrations, P did not limit denitrification rates. In addition to wetting period, residence time, and maintenance of anaerobic conditions, the availability of labile DOC is playing an important limiting role in sediment denitrification within mitigation wetlands.

Evaluation of the organophosphorus hydrolase enzyme activity in creams and investigation of its stability

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Mariye Rajaie

2016-06-01

Full Text Available The main purpose of this project is investigation of the organophosphorus hydrolase (OPH enzyme activity in water in oil (w/o and oil in water (o/w creams and investigation of the OPH enzyme stability in formulated creams. OPH enzyme was extracted and purified from strain flavobacterium. The w/o and o/w creams were prepared using different formulations. In order to achieve an emulsion with maximum stability, appropriate percentage of the cream components was selected by studying different formulations and the physical and chemical stability of the produced cream were considered. 5Uenzyme/90gcream enzyme was used for each formulation. To measure the enzyme activity in creams, extraction method was used and enzyme activity was determined based on parathion hydrolysis. The thermal stability of OPH in both types of w/o and o/w creams was studied at 4 and 30  °C for various time periods. The average enzyme activity was about 0.0065 U/gcream and 0.018 U/gcream for w/o and o/w creams respectivly. According to the results, the relative activity at 4 °C was reduced to 50% after 26 and 45 days in w/o and o/w creams, respectivly. The results showed that the OPH enzyme activity in o/w cream was 2.6 times more than that of w/o cream, because of the higher hydrophobicity of o/w cream compared to w/o. The OPH enzyme stability in o/w cream was greater in comparison to w/o cream. The OPH enzyme was active for nearly 2 months on o/w creams at 4 °C .

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

Science.gov (United States)

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

2014-05-01

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

Seasonal variation in the temperature sensitivity of proteolytic enzyme activity in temperate forest soils

Science.gov (United States)

Brzostek, Edward R.; Finzi, Adrien C.

2012-03-01

Increasing soil temperature has the potential to alter the activity of the extracellular enzymes that mobilize nitrogen (N) from soil organic matter (SOM) and ultimately the availability of N for primary production. Proteolytic enzymes depolymerize N from proteinaceous components of SOM into amino acids, and their activity is a principal driver of the within-system cycle of soil N. The objectives of this study were to investigate whether the soils of temperate forest tree species differ in the temperature sensitivity of proteolytic enzyme activity over the growing season and the role of substrate limitation in regulating temperature sensitivity. Across species and sampling dates, proteolytic enzyme activity had relatively low sensitivity to temperature with a mean activation energy (Ea) of 33.5 kJ mol-1. Ea declined in white ash, American beech, and eastern hemlock soils across the growing season as soils warmed. By contrast, Eain sugar maple soil increased across the growing season. We used these data to develop a species-specific empirical model of proteolytic enzyme activity for the 2009 calendar year and studied the interactive effects of soil temperature (ambient or +5°C) and substrate limitation (ambient or elevated protein) on enzyme activity. Declines in substrate limitation had a larger single-factor effect on proteolytic enzyme activity than temperature, particularly in the spring. There was, however, a large synergistic effect of increasing temperature and substrate supply on proteolytic enzyme activity. Our results suggest limited increases in N availability with climate warming unless there is a parallel increase in the availability of protein substrates.

Evaluation of gasoline-denatured ethanol as a carbon source for denitrification.

Science.gov (United States)

Kazasi, Anna; Boardman, Gregory D; Bott, Charles B

2013-06-01

In this study concerning denitrification, the performance of three carbon sources, methanol (MeOH), ethanol (EtOH) and gasoline-denatured ethanol (dEtOH), was compared and evaluated on the basis of treatment efficiency, inhibition potential and cost. The gasoline denaturant considered here contained mostly aliphatic compounds and little of the components that typically boost the octane rating, such as benzene, toluene, ethylbenzene and xylenes. Results were obtained using three lab-scale SBRs operated at SRT of 12.0 +/- 0.9 days. After biomass was acclimated, denitrification rates with dEtOH were similar to those of EtOH (201 +/- 50 and 197 +/- 28 NO3-N/g MLVSS x d, respectively), and higher than those of MeOH (165 +/- 49 mg NO3-N/g MLVSS x d). The denaturant did not affect biomass production, nitrification or denitrification. Effluent soluble COD concentrations were always less than the analytical detection limit. Although the cost of dEtOH ($2.00/kg nitrate removed) was somewhat higher than that of methanol ($1.63/kg nitrate removed), the use of dEtOH is very promising and utilities will have to decide if it is worth paying a little extra to take advantage of its benefits.

Arabinogalactan proteins: focus on carbohydrate active enzymes

Directory of Open Access Journals (Sweden)

Eva eKnoch

2014-06-01

Full Text Available Arabinogalactan proteins (AGPs are a highly diverse class of cell surface proteoglycans that are commonly found in most plant species. AGPs play important roles in many cellular processes during plant development, such as reproduction, cell proliferation, pattern formation and growth, and in plant-microbe interaction. However, little is known about the molecular mechanisms of their function. Numerous studies using monoclonal antibodies that recognize different AGP glycan epitopes have shown the appearance of a slightly altered AGP glycan in a specific stage of development in plant cells. Therefore, it is anticipated that the biosynthesis and degradation of AGP glycan is tightly regulated during development. Until recently, however, little was known about the enzymes involved in the metabolism of AGP glycans. In this review, we summarize recent discoveries of carbohydrate active enzymes (CAZy; http://www.cazy.org/ involved in the biosynthesis and degradation of AGP glycans, and we discuss the biological role of these enzymes in plant development.

Enzyme activity of a Phanerochaete chrysosporium cellobiohydrolase

African Journals Online (AJOL)

The aim of this study was to produce a secreted, heterologously expressed Phanerochaete chrysosporium cellobiohydrolase (CBHI.1) protein that required no in vitro chemical refolding and to investigate the cellulolytic activity of the clone expressing the glutathione S-transferase (GST) fused CBHI.1 protein. Plate enzyme ...

Enzyme activity in bioregulator-treated tomato (Solanum ...

African Journals Online (AJOL)

USER

2010-05-31

May 31, 2010 ... African Journal of Biotechnology Vol. 9(22), pp. 3264-3271, 31 ... In this work, spectrophotometric analysis ... most stable enzymes in vegetables and its thermal destruc-tion ... proteins, carbohydrates, lipids and allelochemicals (Hedin et al., 1995) ..... activities isolated from corn root plasma membrane. Plant.

The feasibility of enzyme targeted activation for amino acid/dipeptide monoester prodrugs of floxuridine; cathepsin D as a potential targeted enzyme.

Science.gov (United States)

Tsume, Yasuhiro; Amidon, Gordon L

2012-03-26

The improvement of therapeutic efficacy for cancer agents has been a big challenge which includes the increase of tumor selectivity and the reduction of adverse effects at non-tumor sites. In order to achieve those goals, prodrug approaches have been extensively investigated. In this report, the potential activation enzymes for 5'-amino acid/dipeptide monoester floxuridine prodrugs in pancreatic cancer cells were selected and the feasibility of enzyme specific activation of prodrugs was evaluated. All prodrugs exhibited the range of 3.0-105.7 min of half life in Capan-2 cell homogenate with the presence and the absence of selective enzyme inhibitors. 5'-O-L-Phenylalanyl-L-tyrosyl-floxuridine exhibited longer half life only with the presence of pepstatin A. Human cathepsin B and D selectively hydrolized 5'-O-L-phenylalanyl-L-tyrosylfloxuridine and 5'-O-L-phenylalanyl-L-glycylfloxuridine compared to the other tested prodrugs. The wide range of growth inhibitory effect by floxuridine prodrugs in Capan-2 cells was observed due to the different affinities of prodrug promoieties to enzymes. In conclusion, it is feasible to design prodrugs which are activated by specific enzymes. Cathepsin D might be a good candidate as a target enzyme for prodrug activation and 5'-O-L-phenylalanyl-L-tyrosylfloxuridine may be the best candidate among the tested floxuridine prodrugs.

Effect of Robola and Cabernet Sauvignon extracts on platelet activating factor enzymes activity on U937 cells.

Science.gov (United States)

Xanthopoulou, M N; Asimakopoulos, D; Antonopoulou, S; Demopoulos, C A; Fragopoulou, E

2014-12-15

A number of studies support the anti-atherogenic effect of wine compounds. The scope of this study was to examine the effect of a red (Cabernet Sauvignon-CS) and a white (Robola-R) wine, as well as resveratrol and quercetin, on the platelet activating factor (PAF) biosynthetic enzymes, acetyl-CoA:lyso-PAF acetyltransferase (lyso-PAF-AT) and DTT-insensitive CDP-choline 1-alkyl-2-acetyl-sn-glycerol cholinephosphotransferase (PAF-CPT), and its main catabolic enzyme (PAF acetylhydrolase; PAF-AH), on U937 cells, in cell free and in intact cell experiments. In cell free experiments, phenolic compounds and wine extracts inhibited PAF biosynthetic enzymes, however in higher concentrations than intact cell experiments. In the latter cases, polar lipids of both wines inhibited in the same order of magnitude the action of lyso-PAF-AT and of PAF-CPT. The water fractions possessed a dual action, in lower concentrations they activated both enzymes, while in higher concentrations only inhibited PAF-CPT. All fractions either did not affect or slightly activated PAF-AH activity. In conclusion, wine compounds may exert their anti-inflammatory activity by reducing PAF levels through modulation of the PAF metabolic enzymes. Copyright © 2014 Elsevier Ltd. All rights reserved.

Assessment of denitrification gaseous end-products in the soil profile under two water table management practices using repeated measures analysis.

Science.gov (United States)

Elmi, Abdirashid A; Astatkie, Tess; Madramootoo, Chandra; Gordon, Robert; Burton, David

2005-01-01

The denitrification process and nitrous oxide (N2O) production in the soil profile are poorly documented because most research into denitrification has concentrated on the upper soil layer (0-0.15 m). This study, undertaken during the 1999 and 2000 growing seasons, was designed to examine the effects of water table management (WTM), nitrogen (N) application rate, and depth (0.15, 0.30, and 0.45 m) on soil denitrification end-products (N2O and N2) from a corn (Zea mays L.) field. Water table management treatments were free drainage (FD) with open drains and subirrigation (SI) with a target water table depth of 0.6 m. Fertility treatments (ammonium nitrate) were 120 kg N ha(-1) (N120) and 200 kg N ha(-1) (N200). During both growing seasons greater denitrification rates were measured in SI than in FD, particularly in the surface soil (0-0.15 m) and at the intermediate (0.15-0.30 m) soil depths under N200 treatment. Greater denitrification rates under the SI treatment, however, were not accompanied with greater N2O production. The decrease in N2O production under SI was probably caused by a more complete reduction of N2O to N2, which resulted in lower N2O to (N2O + N2) ratios. Denitrification rate, N2O production and N2O to (N2O + N2) ratios were only minimally affected by N treatments, irrespective of sampling date and soil depth. Overall, half of the denitrification occurred at the 0.15- to 0.30- and 0.30- to 0.45-m soil layers, and under SI, regardless of fertility treatment level. Consequently, sampling of the 0- to 0.15-m soil layer alone may not give an accurate estimation of denitrification losses under SI practice.

Effect of soil saturation on denitrification in a grassland soil

Directory of Open Access Journals (Sweden)

L. M. Cardenas

2017-10-01

Full Text Available Nitrous oxide (N2O is of major importance as a greenhouse gas and precursor of ozone (O3 destruction in the stratosphere mostly produced in soils. The soil-emitted N2O is generally predominantly derived from denitrification and, to a smaller extent, nitrification, both processes controlled by environmental factors and their interactions, and are influenced by agricultural management. Soil water content expressed as water-filled pore space (WFPS is a major controlling factor of emissions and its interaction with compaction, has not been studied at the micropore scale. A laboratory incubation was carried out at different saturation levels for a grassland soil and emissions of N2O and N2 were measured as well as the isotopocules of N2O. We found that flux variability was larger in the less saturated soils probably due to nutrient distribution heterogeneity created from soil cracks and consequently nutrient hot spots. The results agreed with denitrification as the main source of fluxes at the highest saturations, but nitrification could have occurred at the lower saturation, even though moisture was still high (71 % WFSP. The isotopocules data indicated isotopic similarities in the wettest treatments vs. the two drier ones. The results agreed with previous findings where it is clear there are two N pools with different dynamics: added N producing intense denitrification vs. soil N resulting in less isotopic fractionation.

Enzyme activity and allosteric characteristics of gamma-irradiated solid aspartate transcarbamylase

International Nuclear Information System (INIS)

Bigler, W.N.; Tolbert, B.M.

1977-01-01

Aspartate transcarbamylase purified from E. coli was lyophilized, irradiated in vacuo with γ radiation from a cesium-137 source, redissolved in buffer under a nitrogen atmosphere, and assayed for enzyme activity. Lyophilized and redissolved enzyme had normal catalytic and allosteric kinetic characteristics. The average D 37 observed with saturating substrate, 25 mM aspartate, was 4.1 Mrad. With less than saturating substrate, 5 mM aspartate, the activity increases from zero to 1.6 Mrad and then decreases with a D 37 of 7.2 Mrad. Inclusion of 1 mM CTP, an allosteric inhibitor, in the 5 mM aspartate assays results in a more pronounced maximum in the activity curve occurring at slightly higher dose, 2.2 Mrad. Inhibitability by CTP has a D 37 of 2.3 Mrad with doses below the activity maximum. Enzyme lyophilized in the presence of 1 mM CTP has a D 37 of 2.9 Mrad. ATCase activity changes caused by irradiation of lyophylized bacteria were qualitatively like the changes observed in the detailed studies with the purified enzyme. Apparent radiation sensitivities of ATCase in lyophilized bacteria were observed to vary with the technique used to disrupt the resuspended bacteria

The effect of millennial-scale changes in Arabian Sea denitrification on atmospheric CO2

International Nuclear Information System (INIS)

Altabet, M.A.; Higginson, M.J.; Murray, D.W.

2002-01-01

Most global biogeochemical processes are known to respond to climate change, some of which have the capacity to produce feedbacks through the regulation of atmospheric greenhouse gases. Marine denitrification - the reduction of nitrate to gaseous nitrogen - is an important process in this regard, affecting greenhouse gas concentrations directly through the incidental production of nitrous oxide, and indirectly through modification of the marine nitrogen inventory and hence the biological pump for C0 2 . Although denitrification has been shown to vary with glacial-interglacial cycles, its response to more rapid climate change has not yet been well characterized. Here we present nitrogen isotope ratio, nitrogen content and chlorin abundance data from sediment cores with high accumulation rates on the Oman continental margin that reveal substantial millennial-scale variability in Arabian Sea denitrification and productivity during the last glacial period. The detailed correspondence of these changes with Dansgaard-Oeschger events recorded in Greenland ice cores indicates rapid, century-scale reorganization of the Arabian Sea ecosystem in response to climate excursions, mediated through the intensity of summer monsoonal upwelling. Considering the several-thousand-year residence time of fixed nitrogen in the ocean, the response of global marine productivity to changes in denitrification would have occurred at lower frequency and appears to be related to climatic and atmospheric C0 2 oscillations observed in Antarctic ice cores between 20 and A kyr ago. (author)

Disturbances in lysosomal enzymes activity in rats, following experimental postradiation disease

International Nuclear Information System (INIS)

Drozdz, M.; Piwowarczyk, B.; Olczyk, K.; Pikula-Zachara, M.

1981-01-01

The studies were aimed at detecting the biological effects of radiation on rat's organism, through studying the activity of lysosomal enzymes in blood plasma and some organs. The contemporary studies suggest that lysosomes play an important role in the occurrence and course of postradiation disease. The obtained results suggest the multidirectional gamma-rays effects on lysosomal enzymes response in serum, leucocytes, liver lysosomes and in liver, kidneys, lungs, heart. Increased activity of acid phosphatase, beta-glucoronidase and beta-acetyl-glucosaminase in the tissues of irradiated animals indicates that gamma rays labilizate the lysosomal membrane. The range of changes indicates a selective nature of this phenomenon. Kidneys, lungs and liver appeared the most ray-sensitive organs. The activity of acid phosphatase was found to be most increased in blood serum and leucocytes. The activity of all examined enzymes in liver lysosomes was decreased. Acid phosphatase exhibited the greatest activity increases. Lysosomal responses are indicative of the degree of destructive or regenerative changes in the organism. (author)

Characterization of inhibitor(s) of β-glucuronidase enzyme activity in GUS-transgenic wheat

KAUST Repository

Ramadan, Ahmed M Ali

2011-06-26

The uidA gene, encoding for β-glucuronidase (GUS), is the most frequently used reporter gene in plants. As a reporter enzyme, GUS can be assayed both qualitatively and quantitatively. In wheat, there are numerous reports of failure in detecting GUS enzyme activity in tissues of transgenic plants, while other reports have suggested presence of β-glucuronidase inhibitor(s) in wheat tissues. In the present study, we show that the β-glucuronidase enzyme activity is not only tissue-specific but also genotype-dependent. Our data demonstrate that the glucuronic acid could be the candidate inhibitor for β-glucuronidase enzyme activity in wheat leaves and roots. It should be noted that the assays to detect β-glucuronidase enzyme activity in wheat should be interpreted carefully. Based on the data of our present study, we recommend studying the chemical pathways, the unintended effects and the possible loss-of-function of any candidate transgene prior to transformation experiments. © 2011 Springer Science+Business Media B.V.

Characterization of inhibitor(s) of β-glucuronidase enzyme activity in GUS-transgenic wheat

KAUST Repository

Ramadan, Ahmed M Ali; Eissa, Hala F.; El-Domyati, Fotouh M.; Saleh, Osama Mesilhy; Ibrahim, Nasser E.; Salama, M. I.; Mahfouz, Magdy M.; Bahieldin, Ahmed M.

2011-01-01

The uidA gene, encoding for β-glucuronidase (GUS), is the most frequently used reporter gene in plants. As a reporter enzyme, GUS can be assayed both qualitatively and quantitatively. In wheat, there are numerous reports of failure in detecting GUS enzyme activity in tissues of transgenic plants, while other reports have suggested presence of β-glucuronidase inhibitor(s) in wheat tissues. In the present study, we show that the β-glucuronidase enzyme activity is not only tissue-specific but also genotype-dependent. Our data demonstrate that the glucuronic acid could be the candidate inhibitor for β-glucuronidase enzyme activity in wheat leaves and roots. It should be noted that the assays to detect β-glucuronidase enzyme activity in wheat should be interpreted carefully. Based on the data of our present study, we recommend studying the chemical pathways, the unintended effects and the possible loss-of-function of any candidate transgene prior to transformation experiments. © 2011 Springer Science+Business Media B.V.

Fungi regulate response of N2O production to warming and grazing in a Tibetan grassland

Science.gov (United States)

Zhong, Lei; Wang, Shiping; Xu, Xingliang; Wang, Yanfen; Rui, Yichao; Zhou, Xiaoqi; Shen, Qinhua; Wang, Jinzhi; Jiang, Lili; Luo, Caiyun; Gu, Tianbao; Ma, Wenchao; Chen, Guanyi

2018-03-01

Lack of understanding of the effects of warming and winter grazing on soil fungal contribution to nitrous oxide (N2O) production has limited our ability to predict N2O fluxes under changes in climate and land use management, because soil fungi play an important role in driving terrestrial N cycling. Here, we examined the effects of 10 years' warming and winter grazing on soil N2O emissions potential in an alpine meadow. Our results showed that soil bacteria and fungi contributed 46 % and 54 % to nitrification, and 37 % and 63 % to denitrification, respectively. Neither warming nor winter grazing affected the activity of enzymes responsible for overall nitrification and denitrification. However, warming significantly increased the enzyme activity of bacterial nitrification and denitrification to 53 % and 55 %, respectively. Warming significantly decreased enzyme activity of fungal nitrification and denitrification to 47 % and 45 %, respectively, while winter grazing had no such effect. We conclude that soil fungi could be the main source for N2O production potential in the Tibetan alpine grasslands. Warming and winter grazing may not affect the potential for soil N2O production potential, but climate warming can alter biotic pathways responsible for N2O production. These findings indicate that characterizing how fungal nitrification/denitrification contributes to N2O production, as well as how it responds to environmental and land use changes, can advance our understanding of N cycling. Therefore, our results provide some new insights about ecological controls on N2O production and lead to refine greenhouse gas flux models.

Antibacterial and glucosyltransferase enzyme inhibitory activity of helmyntostachyszelanica

Science.gov (United States)

Kuspradini, H.; Putri, AS; Mitsunaga, T.

2018-04-01

Helminthostachyszeylanica is a terrestrial, herbaceous, fern-like plant of southeastern Asia and Australia, commonly known as tunjuk-langit. This kind of plant have a medicinal properties such as treatment of malaria, dysentery and can be eaten with betel in the treatment of whooping cough. To evaluate the scientific basis for the use of the plant, the antimicrobial activities of extracts of the stem and leaves were evaluated. The bacteria used in this study is Streptococcus sobrinus, a species of gram-positive, that may be associated with human dental caries. The dried powdered plant parts were extracted using methanol and 50% aqueous extract and screened for their antibacterial effects of Streptococcus sobrinus using the 96 well-plate microdilution broth method. The inhibitory activities of its related enzyme were also determined. The plant extracts showed variable antibacterial and Glucosyltransferase enzyme inhibitory activity while some extracts could not cause any inhibition. It was shown that 50% ethanolics of Helminthostachyzeylanica stem have a potency as anti dental caries agents.

Selective enhancement and verification of woody biomass digestibility as a denitrification carbon source.

Science.gov (United States)

Hu, Rongting; Zheng, Xilai; Xin, Jia; Sun, Zhaoyue; Zheng, Tianyuan

2017-11-01

The denitrification efficiency of woody biomass as carbon source is low because of its poor carbon availability. In this study, representative poplar sawdust was pretreated with lime and peracetic acid to enhance the biomass digestibility to different degrees; sawdust was then mixed with soil to investigate its denitrification efficiency. Under controllable conditions (25-95°C, 12-24h, varying dosages), sawdust digestibility (characterized by reducing sugar yield) was selectively enhanced 1.0-21.8 times over that of the raw sawdust (28.8mgeq.glucoseg -1 dry biomass). This increase was mainly attributed to the removal of lignin from the biomass. As a carbon source, the sawdust (digestibility enhanced by 5.4 times) increased the nitrate removal rate by 4.7 times, without N 2 O emission. However, the sawdust with high digestibility (12.6 or 18.0 times), despite releasing more dissolved organic carbon (DOC), did not exhibit further increase in denitrification efficiency, and emitted N 2 O. Copyright © 2017 Elsevier Ltd. All rights reserved.

Characteristic of nitrous oxide production in partial denitrification process with high nitrite accumulation.

Science.gov (United States)

Du, Rui; Peng, Yongzhen; Cao, Shenbin; Wang, Shuying; Niu, Meng

2016-03-01

Nitrous oxide (N2O) production during the partial denitrification process with nitrate (NO3(-)-N) to nitrite (NO2(-)-N) transformation ratio of 80% was investigated in this study. Results showed that N2O was seldom observed before complete depletion of NO3(-)-N, but it was closely related to the reduction of NO2(-)-N rather than NO3(-)-N. High COD/NO3(-)-N was in favor of N2O production in partial denitrification with high NO2(-)-N accumulation. It was seriously enhanced at constant acidic pH due to the free nitrous acid (FNA) inhibition. However, the N2O production was much lower at initial pH of 5.5 and 6.5 due to the pH increase during denitrification process. Significantly, the pH turning point could be chosen as a controlled parameter to denote the end of NO3(-)-N reduction, which could not only achieve high NO2(-)-N accumulation but also decrease the N2O production significantly for practical application. Copyright © 2015 Elsevier Ltd. All rights reserved.

Optimization of ultrasound-assisted extraction of pectinase enzyme from guava (Psidium guajava) peel: Enzyme recovery, specific activity, temperature, and storage stability.

Science.gov (United States)

Amid, Mehrnoush; Murshid, Fara Syazana; Manap, Mohd Yazid; Islam Sarker, Zaidul

2016-01-01

This study aimed to investigate the effects of the ultrasound-assisted extraction conditions on the yield, specific activity, temperature, and storage stability of the pectinase enzyme from guava peel. The ultrasound variables studied were sonication time (10-30 min), ultrasound temperature (30-50 °C), pH (2.0-8.0), and solvent-to-sample ratio (2:1 mL/g to 6:1 mL/g). The main goal was to optimize the ultrasound-assisted extraction conditions to maximize the recovery of pectinase from guava peel with the most desirable enzyme-specific activity and stability. Under the optimum conditions, a high yield (96.2%), good specific activity (18.2 U/mg), temperature stability (88.3%), and storage stability (90.3%) of the extracted enzyme were achieved. The optimal conditions were 20 min sonication time, 40 °C temperature, at pH 5.0, using a 4:1 mL/g solvent-to-sample ratio. The study demonstrated that optimization of ultrasound-assisted process conditions for the enzyme extraction could improve the enzymatic characteristics and yield of the enzyme.

An easy and efficient permeabilization protocol for in vivo enzyme activity assays in cyanobacteria

DEFF Research Database (Denmark)

Rasmussen, Randi Engelberth; Erstad, Simon Matthé; Ramos Martinez, Erick Miguel

2016-01-01

microbial cell factories. Better understanding of the activities of enzymes involved in the central carbon metabolism would lead to increasing product yields. Currently cell-free lysates are the most widely used method for determination of intracellular enzyme activities. However, due to thick cell walls...... used directly in the assays, the permeabilized cells exhibited the enzyme activities that are comparable or even higher than those detected for cell-free lysates. Moreover, the permeabilized cells could be stored at -20 °C without losing the enzyme activities. The permeabilization process...... for permeabilization of the cyanobacteria Synechococcus sp. PCC 7002 and Synechocystis sp. PCC 6803, and determination of two intracellular enzymes, ribulose-1,5-bisphosphate carboxylase/decarboxylase (Rubisco) and glucose-6-phosphate dehydrogenase (G6PDH), that play pivotal roles in the central carbon metabolism...

A soil infiltration system incorporated with sulfur-utilizing autotrophic denitrification (SISSAD) for domestic wastewater treatment.

Science.gov (United States)

Kong, Zhe; Feng, Chuanping; Chen, Nan; Tong, Shuang; Zhang, Baogang; Hao, Chunbo; Chen, Kun

2014-05-01

To enhance the denitrification performance of soil infiltration, a soil infiltration system incorporated with sulfur-utilizing autotrophic denitrification (SISSAD) for domestic wastewater treatment was developed, and the SISSAD performance was evaluated using synthetic domestic wastewater in this study. The aerobic respiration and nitrification were mainly taken place in the upper aerobic stage (AES), removed 88.44% COD and 89.99% NH4(+)-N. Moreover, autotrophic denitrification occurred in the bottom anaerobic stage (ANS), using the CO2 produced from AES as inorganic carbon source. Results demonstrated that the SISSAD showed a remarkable performance on COD removal efficiency of 95.09%, 84.86% for NO3(-)-N, 95.25% for NH4(+)-N and 93.15% for TP. This research revealed the developed system exhibits a promising application prospect for domestic wastewater in the future. Copyright © 2014 Elsevier Ltd. All rights reserved.

Evidence for denitrification as main source of N2O emission from residue-amended soil

DEFF Research Database (Denmark)

Li, Xiaoxi; Sørensen, Peter; Olesen, Jørgen Eivind

2016-01-01

-leguminous species (ryegrass). Plant material was placed in a discrete layer surrounded by soil in which the nitrate View the MathML source pool was enriched with 15N to distinguish N2O derived from denitrification and nitrification. Net N mineralisation from leguminous catch crops was significant (30–48 mg N kg−1....... Emission of N2O occurred at all moisture levels, but was higher at 50 and 60% WFPS than at 40% in soil with leguminous residues. The 15N enrichment of N2O indicated that denitrification was the dominant source independent of moisture level and residue type. We conclude that catch crop residues...... will stimulate N2O emissions via denitrification over a wide range of soil moisture conditions, but that emission levels may depend significantly on residue quality and soil moisture....

Measurement of peroxisomal enzyme activities in the liver of brown trout (Salmo trutta, using spectrophotometric methods

Directory of Open Access Journals (Sweden)

Resende Albina D

2003-03-01

Full Text Available Abstract Background This study was aimed primarily at testing in the liver of brown trout (Salmo trutta spectrophotometric methods previously used to measure the activities of catalase and hydrogen peroxide producing oxidases in mammals. To evaluate the influence of temperature on the activities of those peroxisomal enzymes was the second objective. A third goal of this work was the study of enzyme distribution in crude cell fractions of brown trout liver. Results The assays revealed a linear increase in the activity of all peroxisomal enzymes as the temperature rose from 10° to 37°C. However, while the activities of hydrogen peroxide producing oxidases were strongly influenced by temperature, catalase activity was only slightly affected. A crude fraction enriched with peroxisomes was obtained by differential centrifugation of liver homogenates, and the contamination by other organelles was evaluated by the activities of marker enzymes for mitochondria (succinate dehydrogenase, lysosomes (aryl sulphatase and microsomes (NADPH cytochrome c reductase. For peroxisomal enzymes, the activities per mg of protein (specific activity in liver homogenates were strongly correlated with the activities per g of liver and with the total activities per liver. These correlations were not obtained with crude peroxisomal fractions. Conclusions The spectrophotometric protocols originally used to quantify the activity of mammalian peroxisomal enzymes can be successfully applied to the study of those enzymes in brown trout. Because the activity of all studied peroxisomal enzymes rose in a linear mode with temperature, their activities can be correctly measured between 10° and 37°C. Probably due to contamination by other organelles and losses of soluble matrix enzymes during homogenisation, enzyme activities in crude peroxisomal fractions do not correlate with the activities in liver homogenates. Thus, total homogenates will be used in future seasonal and

Particulate Pyrite Autotrophic Denitrification (PPAD) for Remediation of Nitrate-contaminated Groundwater

Science.gov (United States)

Tong, S.; Rodriguez-Gonzalez, L. C.; Henderson, M.; Feng, C.; Ergas, S. J.

2015-12-01

The rapid movement of human civilization towards urbanization, industrialization, and increased agricultural activities has introduced a large amount of nitrate into groundwater. Nitrate is a toxic substance discharged from groundwater to rivers and leads to decreased dissolved oxygen and eutrophication. For this experiment, an electron donor is needed to convert nitrate into non-toxic nitrogen gas. Pyrite is one of the most abundant minerals in the earth's crust making it an ideal candidate as an electron donor. The overall goal of this research was to investigate the potential for pyrite to be utilized as an electron donor for autotrophic denitrification of nitrate-contaminated groundwater. Batch studies of particulate pyrite autotrophic denitrification (PPAD) of synthetic groundwater (100 mg NO3--N L-1) were set up with varying biomass concentration, pyrite dose, and pyrite particle size. Reactors were seeded with mixed liquor volatile suspended solids (VSS) from a biological nitrogen removal wastewater treatment facility. PPAD using small pyrite particles (exhibited substantial nitrate removal rate, lower sulfate accumulation (5.46 mg SO42-/mg NO3--N) and lower alkalinity consumption (1.70 mg CaCO3/mg NO3--N) when compared to SOD (7.54 mg SO42-/mg NO3--N, 4.57 mg CaCO3/mg NO3--N based on stoichiometric calculation). This research revealed that the PPAD process is a promising technique for nitrate-contaminated groundwater treatment and promoted the utilization of pyrite in the field of environmental remediation.

Seasonality of fibrolytic enzyme activity in herbivore microbial ...

African Journals Online (AJOL)

2012-08-21

Aug 21, 2012 ... liberating end-products such as volatile fatty acids. Cellulase enzyme ... All the other common chemicals such as glacial acetic acid, sodium azide .... specific activity was observed among animal species and between seasons ...

Microbial enzyme activities of peatland soils in south central Alaska lowlands

Science.gov (United States)

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

Changes in growth, survival and digestive enzyme activities of Asian ...

African Journals Online (AJOL)

A study was conducted to determine the effects of different dietary treatments on the growth, survival and digestive enzyme activities of Mystus nemurus larvae. Newly hatched larvae were reared for 14 days in twelve 15 L glass aquaria (for growth and survival) and eight 300 L fiberglass tanks (for enzyme samples) at a ...

Early bichemical markers of effects: Enzyme induction, oncogene activation and markers of oxidative damage

DEFF Research Database (Denmark)

Poulsen, Henrik E.; Loft, Steffen

1995-01-01

Early bichemical marker, enzyme induction, oncogene activation, oxidative damage, low-density lipoprotein......Early bichemical marker, enzyme induction, oncogene activation, oxidative damage, low-density lipoprotein...

Improving enzymatic activities and thermostability of a tri-functional enzyme with SOD, catalase and cell-permeable activities.

Science.gov (United States)

Luangwattananun, Piriya; Eiamphungporn, Warawan; Songtawee, Napat; Bülow, Leif; Isarankura Na Ayudhya, Chartchalerm; Prachayasittikul, Virapong; Yainoy, Sakda

2017-04-10

Synergistic action of major antioxidant enzymes, e.g., superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) is known to be more effective than the action of any single enzyme. Recently, we have engineered a tri-functional enzyme, 6His-MnSOD-TAT/CAT-MnSOD (M-TAT/CM), with SOD, CAT and cell-permeable activities. The protein actively internalized into the cells and showed superior protection against oxidative stress-induced cell death over native enzymes fused with TAT. To improve its molecular size, enzymatic activity and stability, in this study, MnSOD portions of the engineered protein were replaced by CuZnSOD, which is the smallest and the most heat resistant SOD isoform. The newly engineered protein, CAT-CuZnSOD/6His-CuZnSOD-TAT (CS/S-TAT), had a 42% reduction in molecular size and an increase in SOD and CAT activities by 22% and 99%, respectively. After incubation at 70°C for 10min, the CS/S-TAT retained residual SOD activity up to 54% while SOD activity of the M-TAT/CM was completely abolished. Moreover, the protein exhibited a 5-fold improvement in half-life at 70°C. Thus, this work provides insights into the design and synthesis of a smaller but much more stable multifunctional antioxidant enzyme with ability to enter mammalian cells for further application as protective/therapeutic agent against oxidative stress-related conditions. Copyright © 2017 Elsevier B.V. All rights reserved.

Extracellular enzyme activity in a willow sewage treatment system.

Science.gov (United States)

Brzezinska, Maria Swiontek; Lalke-Porczyk, Elżbieta; Kalwasińska, Agnieszka

2012-12-01

This paper presents the results of studies on the activity of extra-cellular enzymes in soil-willow vegetation filter soil which is used in the post-treatment of household sewage in an onsite wastewater treatment system located in central Poland. Wastewater is discharged from the detached house by gravity into the onsite wastewater treatment system. It flows through a connecting pipe into a single-chamber septic tank and is directed by the connecting pipe to a control well to be further channelled in the soil-willow filter by means of a subsurface leaching system. Soil samples for the studies were collected from two depths of 5 cm and 1 m from three plots: close to the wastewater inflow, at mid-length of the plot and close to its terminal part. Soil samples were collected from May to October 2009. The activity of the extra-cellular enzymes was assayed by the fluorometric method using 4-methylumbelliferyl and 7-amido-4-methylcoumarin substrate. The ranking of potential activity of the assayed enzymes was the same at 5 cm and 1 m soil depths, i.e. esterase > phosphmomoesterase > leucine-aminopeptidase > β-glucosidase > α-glucosidase. The highest values of enzymatic activity were recorded in the surface layer of the soil at the wastewater inflow and decreased with increasing distance from that point.

Enzyme Enzyme activities in relation to sugar accumulation in tomato

International Nuclear Information System (INIS)

Alam, M.J.; Rahman, M.H.; Mamun, M.A.; Islam, K.

2006-01-01

Enzyme activities in tomato juice of five different varieties viz. Ratan, Marglove, BARI-1, BARI-5 and BARI-6, in relation to sugar accumulation were investigated at different maturity stages. The highest amount of invertase and beta-galactosidase was found in Marglove and the lowest in BARI- 6 at all maturity stages. Total soluble sugar and sucrose contents were highest in BARI-1 and lowest in BARI-6. The activity of amylase was maximum in Ratan and minimum in Marglove. Protease activity was highest in Ratan and lowest in BARI-6. BARI-1 contained the highest cellulase activity and the lowest in BARI-5. The amount of total soluble sugar and sucrose increased moderately from premature to ripe stage. The activities of amylase and cellulase increased up to the mature stage and then decreased drastically in the ripe stage. The activities of invertase and protease increased sharply from the premature to the ripe stage while the beta-galactosidase activity decreased remarkably. No detectable amount of reducing sugar was present in the premature stage in all cultivars of tomato but increased thereafter upto the ripe stage. The highest reducing sugar was present in BARI-5 in all of the maturity stages. (author)

Effects of Recurring Droughts on Extracellular Enzyme Activity in Mountain Grassland

Science.gov (United States)

Fuchslueger, L.; Bahn, M.; Kienzl, S.; Hofhansl, F.; Schnecker, J.; Richter, A.

2015-12-01

Water availability is a key factor for biogeochemical processes and determines microbial activity and functioning, and thereby organic matter decomposition in soils by affecting the osmotic potential, soil pore connectivity, substrate diffusion and nutrient availability. Low water availability during drought periods therefore directly affects microbial activity. Recurring drought periods likely induce shifts in microbial structure that might be reflected in altered responses of microbial turnover of organic matter by extracellular enzymes. To study this we measured a set of potential extracellular enzyme activity rates (cellobiohydrolase CBH; leucine-amino-peptidase LAP; phosphatase PHOS; phenoloxidase POX), in grassland soils that were exposed to extreme experimental droughts during the growing seasons of up to five subsequent years. During the first drought period after eight weeks of rain exclusion all measured potential enzyme activities were significantly decreased. In parallel, soil extractable organic carbon and nitrogen concentrations increased and microbial community structure, determined by phospholipid fatty acid analysis, changed. In soils that were exposed to two and three drought periods only PHOS decreased. After four years of drought again CBH, PHOS and POX decreased, while LAP was unaffected; after five years of drought PHOS and POX decreased and CBH and LAP remained stable. Thus, our results suggest that recurring extreme drought events can cause different responses of extracellular enzyme activities and that the responses change over time. We will discuss whether and to what degree these changes were related to shifts in microbial community composition. However, independent of whether a solitary or a recurrent drought was imposed, in cases when enzyme activity rates were altered during drought, they quickly recovered after rewetting. Overall, our data suggest that microbial functioning in mountain grassland is sensitive to drought, but highly

Impact of partial nitritation degree and C/N ratio on simultaneous Sludge Fermentation, Denitrification and Anammox process.

Science.gov (United States)

Wang, Bo; Peng, Yongzhen; Guo, Yuanyuan; Yuan, Yue; Zhao, Mengyue; Wang, Shuying

2016-11-01

This study presents a novel process (i.e. PN/SFDA) to remove nitrogen from low C/N domestic wastewater. The process mainly involves two reactors, a pre-Sequencing Batch Reactor for partial nitritation (termed as PN-SBR) and an anoxic reactor for integrated Denitrification and Anammox with carbon sources produced from Sludge Fermentation (termed as SFDA). During long-term Runs, NO2(-)/NH4(+) ratio (i.e. NO2(-)-N/NH4(+)-N calculated by mole) in the PN-SBR effluent was gradually increased from 0.2 to 37 by extending aerobic duration, meaning that partial nitritation turning to full nitritation could be achieved. Impact of partial nitritation degree on SFDA process was investigated and the result showed that, NO2(-)/NH4(+) ratios between 2 and 10 were appropriate for the co-existence of denitrification and anammox together in the SFDA reactor, and denitrification instead of anammox contributed greater for nitrogen removal. Further batch tests indicated that anammox collaborated well with denitrification at low C/N (1.0 in this study). Copyright © 2016 Elsevier Ltd. All rights reserved.

Warming-induced changes in denitrifier community structure modulate the ability of phototrophic river biofilms to denitrify

Energy Technology Data Exchange (ETDEWEB)

Boulêtreau, Stéphanie, E-mail: stephanie.bouletreau@univ-tlse3.fr [Université de Toulouse, UPS, INP, EcoLab (Laboratoire Ecologie Fonctionnelle et Environnement), 118 route de Narbonne, F-31062 Toulouse (France); CNRS, EcoLab, F-31062 Toulouse (France); Lyautey, Emilie [Université de Toulouse, UPS, INP, EcoLab (Laboratoire Ecologie Fonctionnelle et Environnement), 118 route de Narbonne, F-31062 Toulouse (France); CNRS, EcoLab, F-31062 Toulouse (France); Dubois, Sophie [Université de Bordeaux, EPOC - OASU, UMR 5805, Station Marine d' Arcachon, 2 rue du Professeur Jolyet, 33120 Arcachon (France); Compin, Arthur [Université de Toulouse, UPS, INP, EcoLab (Laboratoire Ecologie Fonctionnelle et Environnement), 118 route de Narbonne, F-31062 Toulouse (France); CNRS, EcoLab, F-31062 Toulouse (France); Delattre, Cécile; Touron-Bodilis, Aurélie [EDF Recherche et Développement, LNHE (Laboratoire National d' Hydraulique et Environnement), 6 quai Watier, F-78401 Chatou (France); Mastrorillo, Sylvain [Université de Toulouse, UPS, INP, EcoLab (Laboratoire Ecologie Fonctionnelle et Environnement), 118 route de Narbonne, F-31062 Toulouse (France); CNRS, EcoLab, F-31062 Toulouse (France); Garabetian, Frédéric [Université de Bordeaux, EPOC - OASU, UMR 5805, Station Marine d' Arcachon, 2 rue du Professeur Jolyet, 33120 Arcachon (France)

2014-01-01

Microbial denitrification is the main nitrogen removing process in freshwater ecosystems. The aim of this study was to show whether and how water warming (+ 2.5 °C) drives bacterial diversity and structuring and how bacterial diversity affects denitrification enzymatic activity in phototrophic river biofilms (PRB). We used water warming associated to the immediate thermal release of a nuclear power plant cooling circuit to produce natural PRB assemblages on glass slides while testing 2 temperatures (mean temperature of 17 °C versus 19.5 °C). PRB were sampled at 2 sampling times during PRB accretion (6 and 21 days) in both temperatures. Bacterial community composition was assessed using ARISA. Denitrifier community abundance and denitrification gene mRNA levels were estimated by q-PCR and qRT-PCR, respectively, of 5 genes encoding catalytic subunits of the denitrification key enzymes. Denitrification enzyme activity (DEA) was measured by the acetylene-block assay at 20 °C. A mean water warming of 2.5 °C was sufficient to produce contrasted total bacterial and denitrifier communities and, therefore, to affect DEA. Indirect temperature effect on DEA may have varied between sampling time, increasing by up to 10 the denitrification rate of 6-day-old PRB and decreasing by up to 5 the denitrification rate of 21-day-old PRB. The present results suggest that indirect effects of warming through changes in bacterial community composition, coupled to the strong direct effect of temperature on DEA already demonstrated in PRB, could modulate dissolved nitrogen removal by denitrification in rivers and streams. - Highlights: •We produced river biofilms in 2 mean temperature conditions: 17 vs 19.5 °C. •We compared their denitrifiers' structuring and functioning in 6d- and 21d-old biofilms. •A difference of 2.5 °C produced contrasted denitrifier communities. •The indirect temperature effect on denitrification activity shifted between biofilm age. �

High inorganic triphosphatase activities in bacteria and mammalian cells: identification of the enzymes involved.

Directory of Open Access Journals (Sweden)

Gregory Kohn

Full Text Available BACKGROUND: We recently characterized a specific inorganic triphosphatase (PPPase from Nitrosomonas europaea. This enzyme belongs to the CYTH superfamily of proteins. Many bacterial members of this family are annotated as predicted adenylate cyclases, because one of the founding members is CyaB adenylate cyclase from A. hydrophila. The aim of the present study is to determine whether other members of the CYTH protein family also have a PPPase activity, if there are PPPase activities in animal tissues and what enzymes are responsible for these activities. METHODOLOGY/PRINCIPAL FINDINGS: Recombinant enzymes were expressed and purified as GST- or His-tagged fusion proteins and the enzyme activities were determined by measuring the release of inorganic phosphate. We show that the hitherto uncharacterized E. coli CYTH protein ygiF is a specific PPPase, but it contributes only marginally to the total PPPase activity in this organism, where the main enzyme responsible for hydrolysis of inorganic triphosphate (PPP(i is inorganic pyrophosphatase. We further show that CyaB hydrolyzes PPP(i but this activity is low compared to its adenylate cyclase activity. Finally we demonstrate a high PPPase activity in mammalian and quail tissue, particularly in the brain. We show that this activity is mainly due to Prune, an exopolyphosphatase overexpressed in metastatic tumors where it promotes cell motility. CONCLUSIONS AND GENERAL SIGNIFICANCE: We show for the first time that PPPase activities are widespread in bacteria and animals. We identified the enzymes responsible for these activities but we were unable to detect significant amounts of PPP(i in E. coli or brain extracts using ion chromatography and capillary electrophoresis. The role of these enzymes may be to hydrolyze PPP(i, which could be cytotoxic because of its high affinity for Ca(2+, thereby interfering with Ca(2+ signaling.

The diversity of the N2O reducers matters for the N2O:N2 denitrification end-product ratio across an annual and a perennial cropping system.

Science.gov (United States)

Domeignoz-Horta, Luiz A; Spor, Aymé; Bru, David; Breuil, Marie-Christine; Bizouard, Florian; Léonard, Joël; Philippot, Laurent

2015-01-01

Agriculture is the main source of terrestrial emissions of N2O, a potent greenhouse gas and the main cause of ozone layer depletion. The reduction of N2O into N2 by microorganisms carrying the nitrous oxide reductase gene (nosZ) is the only biological process known to eliminate this greenhouse gas. Recent studies showed that a previously unknown clade of N2O-reducers was related to the capacity of the soil to act as an N2O sink, opening the way for new strategies to mitigate emissions. Here, we investigated whether the agricultural practices could differently influence the two N2O reducer clades with consequences for denitrification end-products. The abundance of N2O-reducers and producers was quantified by real-time PCR, and the diversity of both nosZ clades was determined by 454 pyrosequencing. Potential N2O production and potential denitrification activity were used to calculate the denitrification gaseous end-product ratio. Overall, the results showed limited differences between management practices but there were significant differences between cropping systems in both the abundance and structure of the nosZII community, as well as in the [rN2O/r(N2O+N2)] ratio. More limited differences were observed in the nosZI community, suggesting that the newly identified nosZII clade is more sensitive than nosZI to environmental changes. Potential denitrification activity and potential N2O production were explained mainly by the soil properties while the diversity of the nosZII clade on its own explained 26% of the denitrification end-product ratio, which highlights the importance of understanding the ecology of this newly identified clade of N2O reducers for mitigation strategies.

Temperature response of denitrification rate and greenhouse gas production in agricultural river marginal wetland soils.

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Bonnett, S A F; Blackwell, M S A; Leah, R; Cook, V; O'Connor, M; Maltby, E

2013-05-01

Soils are predicted to exhibit significant feedback to global warming via the temperature response of greenhouse gas (GHG) production. However, the temperature response of hydromorphic wetland soils is complicated by confounding factors such as oxygen (O2 ), nitrate (NO3-) and soil carbon (C). We examined the effect of a temperature gradient (2-25 °C) on denitrification rates and net nitrous oxide (N2 O), methane (CH4 ) production and heterotrophic respiration in mineral (Eutric cambisol and Fluvisol) and organic (Histosol) soil types in a river marginal landscape of the Tamar catchment, Devon, UK, under non-flooded and flooded with enriched NO3- conditions. It was hypothesized that the temperature response is dependent on interactions with NO3--enriched flooding, and the physicochemical conditions of these soil types. Denitrification rate (mean, 746 ± 97.3 μg m(-2)  h(-1) ), net N2 O production (mean, 180 ± 26.6 μg m(-2)  h(-1) ) and net CH4 production (mean, 1065 ± 183 μg m(-2)  h(-1) ) were highest in the organic Histosol, with higher organic matter, ammonium and moisture, and lower NO3- concentrations. Heterotrophic respiration (mean, 127 ± 4.6 mg m(-2)  h(-1) ) was not significantly different between soil types and dominated total GHG (CO2 eq) production in all soil types. Generally, the temperature responses of denitrification rate and net N2 O production were exponential, whilst net CH4 production was unresponsive, possibly due to substrate limitation, and heterotrophic respiration was exponential but limited in summer at higher temperatures. Flooding with NO3- increased denitrification rate, net N2 O production and heterotrophic respiration, but a reduction in net CH4 production suggests inhibition of methanogenesis by NO3- or N2 O produced from denitrification. Implications for management and policy are that warming and flood events may promote microbial interactions in soil between distinct microbial communities and increase

Nitrous oxide emission from denitrification in stream and river networks

Science.gov (United States)

Beaulieu, J.J.; Tank, J.L.; Hamilton, S.K.; Wollheim, W.M.; Hall, R.O.; Mulholland, P.J.; Peterson, B.J.; Ashkenas, L.R.; Cooper, L.W.; Dahm, Clifford N.; Dodds, W.K.; Grimm, N. B.; Johnson, S.L.; McDowell, W.H.; Poole, G.C.; Maurice, Valett H.; Arango, C.P.; Bernot, M.J.; Burgin, A.J.; Crenshaw, C.L.; Helton, A.M.; Johnson, L.T.; O'Brien, J. M.; Potter, J.D.; Sheibley, R.W.; Sobota, D.J.; Thomas, S.M.

2011-01-01

Nitrous oxide (N2O) is a potent greenhouse gas that contributes to climate change and stratospheric ozone destruction. Anthropogenic nitrogen (N) loading to river networks is a potentially important source of N 2O via microbial denitrification that converts N to N2O and dinitrogen (N2). The fraction of denitrified N that escapes as N2O rather than N2 (i.e., the N2O yield) is an important determinant of how much N2O is produced by river networks, but little is known about the N2O yield in flowing waters. Here, we present the results of whole-stream 15N-tracer additions conducted in 72 headwater streams draining multiple land-use types across the United States. We found that stream denitrification produces N2O at rates that increase with stream water nitrate (NO3-) concentrations, but that production, but does not increase the N2O yield. In our study, most streams were sources of N2O to the atmosphere and the highest emission rates were observed in streams draining urban basins. Using a global river network model, we estimate that microbial N transformations (e.g., denitrification and nitrification) convert at least 0.68 Tg??y -1 of anthropogenic N inputs to N2O in river networks, equivalent to 10% of the global anthropogenic N2O emission rate. This estimate of stream and river N2O emissions is three times greater than estimated by the Intergovernmental Panel on Climate Change.

Denitrification of nitrate waste solutions

International Nuclear Information System (INIS)

Michaels, S.L.; Michel, R.C.; Terpandjian, P.D.; Vora, J.N.

1976-01-01

Bacterial denitrification by Pseudomonas Stutzeri has been chosen as the method for removing nitrate from the effluent stream of the Y-12 uranium purification process. A model was developed to predict bacterial growth and carbon and nitrate depletion during the induction period and steady state operation. Modification of analytical procedures and automatic control of the pH in the reactor are recommended to improve agreement between the prediction of the model and experimental data. An initial carbon-to-nitrogen (C/N) mass ratio of 1.4-1.5 insures adequate population growth during the induction period. Further experiments in batch reactors and in steady state flow reactors are recommended to obtain more reliable kinetic rate constants

Study on the Correlation between Gene Expression and Enzyme Activity of Seven Key Enzymes and Ginsenoside Content in Ginseng in Over Time in Ji'an, China.

Science.gov (United States)

Yin, Juxin; Zhang, Daihui; Zhuang, Jianjian; Huang, Yi; Mu, Ying; Lv, Shaowu

2017-12-11

Panax ginseng is a traditional medicine. Fresh ginseng is one of the most important industries related to ginseng development, and fresh ginseng of varying ages has different medicinal properties. Previous research has not systematically reported the correlation between changes in key enzyme activity with changes in ginsenoside content in fresh ginseng over time. In this study, for the first time, we use ginseng samples of varying ages in Ji'an and systematically reported the changes in the activity of seven key enzymes (HMGR, FPS, SS, SE, DS, CYP450, and GT). We investigated the content of ginsenoside and gene expression of these key enzymes. Ginsenoside content was measured using HPLC. HPLC, GC-MS, and LC-MS were combined to measure the enzyme activity of the key enzymes. Quantitative PCR was used in the investigation of gene expression. By analyzing the correlation between the enzyme activity and the transcription level of the key enzymes with ginsenoside content, we found that DS and GT enzyme activities are significantly correlated with the ginsenoside content in different ages of ginseng. Our findings might provide a new strategy to discriminate between ginseng of different years. Meanwhile, this research provides important information for the in-depth study of ginsenoside biosynthesis.

The Feasibility of Enzyme Targeted Activation for Amino Acid/Dipeptide Monoester Prodrugs of Floxuridine; Cathepsin D as a Potential Targeted Enzyme

Directory of Open Access Journals (Sweden)

Gordon L. Amidon

2012-03-01

Full Text Available The improvement of therapeutic efficacy for cancer agents has been a big challenge which includes the increase of tumor selectivity and the reduction of adverse effects at non-tumor sites. In order to achieve those goals, prodrug approaches have been extensively investigated. In this report, the potential activation enzymes for 5¢-amino acid/dipeptide monoester floxuridine prodrugs in pancreatic cancer cells were selected and the feasibility of enzyme specific activation of prodrugs was evaluated. All prodrugs exhibited the range of 3.0–105.7 min of half life in Capan-2 cell homogenate with the presence and the absence of selective enzyme inhibitors. 5¢-O-L-Phenylalanyl-L-tyrosyl-floxuridine exhibited longer half life only with the presence of pepstatin A. Human cathepsin B and D selectively hydrolized 5¢-O-L-phenylalanyl-L-tyrosylfloxuridine and 5¢-O-L-phenylalanyl-L-glycylfloxuridine compared to the other tested prodrugs. The wide range of growth inhibitory effect by floxuridine prodrugs in Capan-2 cells was observed due to the different affinities of prodrug promoieties to enyzmes. In conclusion, it is feasible to design prodrugs which are activated by specific enzymes. Cathepsin D might be a good candidate as a target enzyme for prodrug activation and 5¢-O-L-phenylalanyl-L-tyrosylfloxuridine may be the best candidate among the tested floxuridine prodrugs.

Characterization of Carbohydrate Active Enzymes Involved in Arabinogalactan Protein Metabolism

DEFF Research Database (Denmark)

Knoch, Eva

and tissues, their functions and synthesis are still poorly understood. The aim of the research presented in the thesis was to characterize carbohydrate active enzymes involved in AGP biosynthesis and modification to gain insights into the biosynthesis of the glycoproteins in plants. Candidate...... glycosyltransferases and glycoside hydrolases were selected based on co-expression profiles from a transcriptomics analysis. Reverse genetics approach on a novel glucuronosyltransferase involved in AGP biosynthesis has revealed that the enzyme activity is required for normal cell elongation in etiolated seedlings....... The enzymatic activity of a hydrolase from GH family 17 was investigated, without successful determination of the activity. Members of hydrolase family 43 appeared to be localized in the Golgi-apparatus, which is also the compartment for glycan biosynthesis. The localization of these glycoside hydrolases...

Deletion of creB in Aspergillus oryzae increases secreted hydrolytic enzyme activity.

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Hunter, A J; Morris, T A; Jin, B; Saint, C P; Kelly, J M

2013-09-01

Aspergillus oryzae has been used in the food and beverage industry for centuries, and industrial strains have been produced by multiple rounds of selection. Targeted gene deletion technology is particularly useful for strain improvement in such strains, particularly when they do not have a well-characterized meiotic cycle. Phenotypes of an Aspergillus nidulans strain null for the CreB deubiquitinating enzyme include effects on growth and repression, including increased activity levels of various enzymes. We show that Aspergillus oryzae contains a functional homologue of the CreB deubiquitinating enzyme and that a null strain shows increased activity levels of industrially important secreted enzymes, including cellulases, xylanases, amylases, and proteases, as well as alleviated inhibition of spore germination on glucose medium. Reverse transcription-quantitative PCR (RT-qPCR) analysis showed that the increased levels of enzyme activity in both Aspergillus nidulans and Aspergillus oryzae are mirrored at the transcript level, indicating transcriptional regulation. We report that Aspergillus oryzae DAR3699, originally isolated from soy fermentation, has a similar phenotype to that of a creB deletion mutant of the RIB40 strain, and it contains a mutation in the creB gene. Collectively, the results for Aspergillus oryzae, Aspergillus nidulans, Trichoderma reesei, and Penicillium decumbens show that deletion of creB may be broadly useful in diverse fungi for increasing production of a variety of enzymes.

Effect of different nutrient supply and other growth factors on the activity of the oxidizing enzymes in plants

Energy Technology Data Exchange (ETDEWEB)

Amberger, A

1960-01-01

Among the plants studied were french beans and peas; the oxidizing enzymes examined were ascorbic acid oxidase, cytochrome oxidase, phenol oxidase, peroxidase and catalase. Increasing the K dosage reduced enzyme activity and raised dry matter contents until at a very high dosage this action was reversed. Both N and P increased enzyme activity and yields. With B high enzyme activity and low dry matter content were both associated with deficiency and toxicity levels. Increasing the Fe dosage led to a rise in both dry matter content and enzyme activity, whereas F depressed yields and raised enzyme activity. Lack of water increased respiration. Light inhibited all enzyme activity.

Cadmium Phytoavailability and Enzyme Activity under Humic Acid Treatment in Fluvo-aquic Soil

Science.gov (United States)

Liu, Borui; Huang, Qing; Su, Yuefeng

2018-01-01

A pot experiment was conducted to investigate the cadmium (Cd) availability to pakchois (Brassica chinensis L.) as well as the enzyme activities in fluvo-aquic soil under humic acid treatment. The results showed that the phytoavailability of Cd in soil decreased gradually as humic acid concentration rose (0 to 12 g·kg-1), while the activities of urease (UE), alkaline phosphatase (ALP) and catalase (CAT) kept increasing (P enzymes due to the Cd pollution. In conclusion, humic acid is effective for the reduction of both Cd phytoavailability and the damage to enzyme activities due to Cd pollution in fluvo-aquic soil

Is there any role of prolidase enzyme activity in the etiology of preeclampsia?

Science.gov (United States)

Pehlivan, Mustafa; Ozün Ozbay, Pelin; Temur, Muzaffer; Yılmaz, Ozgur; Verit, Fatma Ferda; Aksoy, Nurten; Korkmazer, Engin; Üstünyurt, Emin

2017-05-01

To evaluate a relationship between preeclampsia and prolidase enzyme activity. A prospective cohort study of 41 pregnant women diagnosed with preeclampsia and 31 healthy pregnant women as control group was selected at Harran University Hospital Department of Obstetrics and Gynecology. The prolidase enzyme activity was analyzed in maternal and umbilical cord plasma, amniotic fluid and placental and umbilical cord tissues by Chinard method in addition to maternal serum levels of lactate dehydrogenase (LDH), serum glutamate pyruvate transaminase (SGPT) and serum glutamate oxaloacetate transaminase (SGOT). A significant relationship was found between plasma prolidase activity (635 ± 83 U/L) (p  = 0.007), umbilical cord plasma prolidase activity (610 ± 90 U/L) (p = 0.013), amniotic fluid prolidase activity (558 ± 100 U/L) (p  = 0.001), umbilical cord tissue prolidase activity (4248 ± 1675 U/gr protein) (p  = 0.013) and placental tissue prolidase activity (2116 ± 601 U/gr protein) (p  = 0.001) in preeclamptic group when compared to healthy pregnant women. There is a strong correlation between prolidase enzyme activity and preeclampsia. Prolidase enzyme activity may play a role in preeclampsia.

Nitrous oxide emission from denitrification in stream and river networks

Science.gov (United States)

Nitrous oxide (N2O) is a potent greenhouse gas that contributes to climate change and stratospheric ozone destruction. Anthropogenic nitrogen (N) loading to river networks is a potentially important source of N2O via microbial denitrification which converts N to N2O and dinitrog...

Transient competitive complexation in biological kinetic isotope fractionation explains nonsteady isotopic effects: Theory and application to denitrification in soils

Science.gov (United States)

Maggi, Federico; Riley, William J.

2009-12-01

The theoretical formulation of biological kinetic isotope fractionation often assumes first-order or Michaelis-Menten kinetics, the latter solved under the quasi-steady state assumption. Both formulations lead to a constant isotope fractionation factor, therefore they may return incorrect estimations of isotopic effects and misleading interpretations of isotopic signatures when fractionation is not a steady process. We have analyzed the isotopic signature of denitrification in biogeochemical soil systems by Menyailo and Hungate (2006) in which high and variable 15N-N2O enrichment during N2O production and inverse isotope fractionation during N2O consumption could not be explained with first-order kinetics and the Rayleigh equation, or with Michaelis-Menten kinetics. When Michaelis-Menten kinetics were coupled to Monod kinetics to describe biomass and enzyme dynamics, and the quasi-steady state assumption was relaxed, transient Michaelis-Menten-Monod kinetics accurately reproduced the observed concentrations, and variable and inverse isotope fractionations. These results imply a substantial revision in modeling isotopic effects, suggesting that steady state kinetics such as first-order, Rayleigh, and classic Michaelis-Menten kinetics should be superseded by transient kinetics in conjunction with biomass and enzyme dynamics.

Study of a combined heterotrophic and sulfur autotrophic denitrification technology for removal of nitrate in water

International Nuclear Information System (INIS)

Liu Huijuan; Jiang Wei; Wan Dongjin; Qu Jiuhui

2009-01-01

A combined two-step process of heterotrophic denitrification in a fluidized reactor and sulfur autotrophic denitrification processes (CHSAD) was developed for the removal of nitrate in drinking water. In this process, the advantage of high efficiency of heterotrophic denitrification with non-excessive methanol and the advantage of non-pollution of sulfur autotriphic denitrification were integrated in this CHSAD process. And, this CHSAD process had the capacity of pH balance and could control the concentration of SO 4 2- in effluent by adjusting the operation condition. When the influent nitrate was 30 mg NO 3 - -N/L, the reactor could be operated efficiently at the hydraulic retention time (HRT) ranging from 20 to 40 min with C:N ratio (mg CH 3 OH:mg NO 3 - -N) of 2.0 (methanol as carbon source). The nitrate removal was nearly 100% and there was no accumulated nitrite or residual methanol in the effluent. The effluent pH was about 7.5 and the sulfate concentration was lower than 130 mg/L. The maximum volume-loading rate of the reactor was 2.16 kg NO 3 - -N/(m 3 d). The biomass and scanning electron microscopy graphs of biofilm were also analyzed.

Applicability of industrial wastewater as carbon source for denitrification of a sludge dewatering liquor.

Science.gov (United States)

Chen, Jiazhong; Lee, Yoomin; Oleszkiewicz, Jan A

2013-01-01

The applicability of four industrial waste streams from potato processing, canola processing and oil refining, biodiesel production (glycerol), and glycol as substitutes to methanol and ethanol in denitrification of anaerobically digested sludge dewatering liquor (centrate) was evaluated in bench-scale sequencing batch reactors. It was found that glycerol was the best substitute with the specific denitrification rate (SDNR) of 13 mg NO3-N/(g VSS x h) followed by potato processing wastewater at 12mg NO3-N/(g VSS x h). Both substrates produced faster SDNR than methanol's 10mg NO3-N/(g VSS x h); however, they were inferior to ethanol's 17 mg NO3-N/(g VSS x h). Glycol had SDNR of 8 mg NO3-N/(g VSS x h) and demonstrated a very fast acclimation rate, i.e. the response in increased denitrification rate was visible in three days following glycol addition. Canola processing and oil refining wastewater was considered an inappropriate carbon source due to a low SDNR of 5 mg NO3-N/(g VSS x h) and apparent inhibitory effect on nitrification.

Protein Hydrolysis from Catfish Prepared by Papain Enzyme and Antioxidant Activity of Hydrolyzate

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Ace Baehaki

2015-12-01

Full Text Available The objective of this research was to make a protein hydrolysates from catfish (Pangasius pangasius enzymatically using papain enzyme and analyzed the antioxidant activity of protein hydrolysates produced. The research used the method completely randomized design with two replications the treatment were the difference concentration of the papain enzyme (0%, 1%, 2%, 3%, 4%, 5%, and 6%. The parameters of research were antioxidative activity using DPPH (2,2-difenil-1–pikrilhidrazil, protein content, and molecular weight using SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis. The results showed that catfish protein hydrolysates prepared by papain enzyme has antioxidative activity. The highest degree of hydrolysis was 71.98% at enzyme concentration of 6%. Based on the DPPH scavenging method catfish protein hydrolysates has the antioxidative activity with the value 37.85-67.62%. The protein content of catfish protein hydrolysates were 20.86-54.47 mg/ml. The molecular weight of catfish protein hydrolyzates were 11.90-65.20 kDa.

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

African Journals Online (AJOL)

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

Quantifying denitrification in rippled permeable sands through combined flume experiments and modeling

DEFF Research Database (Denmark)

Kessler, Adam J.; Glud, Ronnie N.; Cardenas, M. Bayani

2012-01-01

We measured denitrification in permeable sediments in a sealed flume tank with environmentally representative fluid flow and solute transport behavior using novel measurements. Numerical flow and reactive transport models representing the flume experiments were implemented to provide mechanistic...... insight into the coupled hydrodynamic and biogeochemical processes. There was broad agreement between the model results and experimental data. The model showed that the coupling between nitrification and denitrification was relatively weak in comparison to that in cohesive sediments. This was due...... of permeable sediments with nonmigratory ripples to remove bioavailable nitrogen from coastal ecosystems is lower than that of cohesive sediments. We conclude that while experimental measurements provide a good starting point for constraining key parameters, reactive transport models with realistic kinetic...

Ratio Imaging of Enzyme Activity Using Dual Wavelength Optical Reporters

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Moritz F. Kircher

2002-04-01

Full Text Available The design of near-infrared fluorescent (NIRF probes that are activated by specific proteases has, for the first time, allowed enzyme activity to be imaged in vivo. In the current study, we report on a method of imaging enzyme activity using two fluorescent probes that, together, provide improved quantitation of enzymatic activity. The method employs two chemically similar probes that differ in their degradability by cathepsin B. One probe consists of the NIRF dye Cy5.5 attached to a particulate carrier, a crosslinked iron oxide nanoparticle (CLIO, through cathepsin B cleavable l-arginyl peptides. A second probe consists of Cy3.5 attached to a CLIO through proteolytically resistant d-arginyl peptides. Using mixtures of the two probes, we have shown that the ratio of Cy5.5 to Cy3.5 fluorescence can be used to determine levels of cathepsin B in the environment of nanoparticles with macrophages in suspension. After intravenous injection, tissue fluorescence from the nondegradable Cy3.5–d-arginyl probe reflected nanoparticle accumulation, while fluorescence of the Cy5.5–l-arginyl probe was dependent on both accumulation and activation by cathepsin B. Dual wavelength ratio imaging can be used for the quantitative imaging of a variety of enzymes in clinically important settings, while the magnetic properties of the probes allow their detection by MR imaging.

Tissue and plasma enzyme activities in juvenile green iguanas.

Science.gov (United States)

Wagner, R A; Wetzel, R

1999-02-01

To determine activities of intracellular enzymes in 8 major organs in juvenile green iguanas and to compare tissue and plasma activities. 6 green iguanas iguanas, but high values may not always indicate overt muscle disease. The AMS activity may be specific for the pancreas, but the wide range of plasma activity would likely limit its diagnostic usefulness. Activities of AST and LDH may reflect tissue damage or inflammation, but probably do not reflect damage to specific tissues or organs.

Enzyme activity assays within microstructured optical fibers enabled by automated alignment.

Science.gov (United States)

Warren-Smith, Stephen C; Nie, Guiying; Schartner, Erik P; Salamonsen, Lois A; Monro, Tanya M

2012-12-01

A fluorescence-based enzyme activity assay has been demonstrated within a small-core microstructured optical fiber (MOF) for the first time. To achieve this, a reflection-based automated alignment system has been developed, which uses feedback and piezoelectric actuators to maintain optical alignment. The auto-alignment system provides optical stability for the time required to perform an activity assay. The chosen assay is based on the enzyme proprotein convertase 5/6 (PC6) and has important applications in women's health.

The effect of hyperthermia and radiation on lysosomal enzyme activity of mouse mammary tumours

International Nuclear Information System (INIS)

Barratt, G.M.; Wills, E.D.

1979-01-01

The effects of hyperthermia and radiation have been studied on the acid phosphatase and β-glucuronidase activities in lysosomes of C3H mice mammary tumours and of the spleen. Quantitative histochemical methods have been used. Hyperthermic treatment of both spontaneous and transplanted tumours caused an increase in the activity of both acid phosphatase and β-glucuronase when measured immediately after treatment, but the activities returned to normal after 24 hours. In contrast a radiation dose of 3500 rad did not cause an increase in activity of either enzyme immediately, but a large activation was observed after 24 hr. Combination of hyperthermic and radiation treatment caused increases in enzyme activities which were dependent on the time after treatment. Hyperthermic treatment of the lower body of mice bearing tumours also caused activation of lysosomal enzymes in the spleen. This may be hormone mediated. It is considered that the increased lysosomal enzyme activity observed after hyperthermia may be a consequence of increased permeability of the lysosomal membrane caused by hyperthermia. (author)

[Estimation of adaptive capacities in Magnitogorsk children from the activity of some detoxification enzymes].

Science.gov (United States)

koganova, Z I; Ingel', F I; Antipanova, N A; Legostoeva, T B; Poliakova, O V

2010-01-01

The paper provides the first fragment of a multiparameter study analyzing the influence of environmental pollution, the social and psychological features of a family, and some endogenous factors on genome stability and sensitivity in a developed ferrous metallurgy town. It also gives data on the urine and serum activity of the lysosomal enzyme N-acetyl-b-D-glucosaminidase (NAG) and the serum activity of catalase in an organized contingent of apparently healthy children (n = 178; 6 kindergartens) aged 5-7 years, who live permanently in Magnitogorsk at different distances from the metallurgical works. More than 70% of children selected for examination were found to have average normal levels of activity of the enzymes studied. According to the average levels of enzyme activity, there were only 2 kindergartens (both from the left-bank region). In the children from the left-bank area, enzyme activities varied more greatly, which suggests the higher prevalence of tense adaptation. Correlation analysis revealed association between the children's serum activity of enzymes and some components of snow pollution. It is anticipated that the found changes in serum activities of N-acetyl-beta-D-glucosaminidase and catalase may be determined by individual differences in a child's response to ambient air pollutants.

Denitrification of high strength nitrate waste from a nuclear industry using acclimatized biomass in a pilot scale reactor.

Science.gov (United States)

Dhamole, Pradip B; Nair, Rashmi R; D'Souza, Stanislaus F; Pandit, Aniruddha B; Lele, S S

2015-01-01

This work investigates the performance of acclimatized biomass for denitrification of high strength nitrate waste (10,000 mg/L NO3) from a nuclear industry in a continuous laboratory scale (32 L) and pilot scale reactor (330 L) operated over a period of 4 and 5 months, respectively. Effect of substrate fluctuations (mainly C/NO3-N) on denitrification was studied in a laboratory scale reactor. Incomplete denitrification (95-96 %) was observed at low C/NO3-N (≤2), whereas at high C/NO3-N (≥2.25) led to ammonia formation. Ammonia production increased from 1 to 9 % with an increase in C/NO3-N from 2.25 to 6. Complete denitrification and no ammonia formation were observed at an optimum C/NO3-N of 2.0. Microbiological studies showed decrease in denitrifiers and increase in nitrite-oxidizing bacteria and ammonia-oxidizing bacteria at high C/NO3-N (≥2.25). Pilot scale studies were carried out with optimum C/NO3-N, and sustainability of the process was checked on the pilot scale for 5 months.

A fluorescence-based hydrolytic enzyme activity assay for quantifying toxic effects of Roundup® to Daphnia magna

DEFF Research Database (Denmark)

Ørsted, Michael; Roslev, Peter

2015-01-01

Daphnia magna is a widely used model organism for aquatic toxicity testing. In the present study, we investigated the hydrolytic enzyme activity of D. magna after exposure to toxicant stress. In vivo enzyme activity was quantified using 15 fluorogenic enzyme probes based on 4-methylumbelliferyl...... or 7-amino-4-methylcoumarin. Probing D. magna enzyme activity was evaluated using short-term exposure (24-48 h) to the reference chemical K2Cr2O7, or the herbicide formulation Roundup®. Toxicant induced changes in hydrolytic enzyme activity were compared to changes in mobility (ISO 6341). The results...... showed that hydrolytic enzyme activity was quantifiable as a combination of whole body fluorescence of D. magna, and fluorescence of the surrounding water. Exposure of D. magna to lethal and sublethal concentrations of Roundup® resulted in loss of whole body enzyme activity, and release of cell...

Spatial characterization of proteolytic enzyme activity in the foregut region of the adult necrophagous fly, Protophormia terraenovae.

Science.gov (United States)

Rivers, David B; Acca, Gillian; Fink, Marc; Brogan, Rebecca; Schoeffield, Andrew

2014-08-01

The spatial distribution of proteolytic enzymes in the adult foregut of Protophormia terraenovae was studied in the context of protein digestion and regurgitation. Based on substrate specificity, pH optima, and use of specific protease inhibitors, all adults tested displayed enzyme activity in the foregut consistent with pepsin, trypsin and chymotrypsin. Chymotrypsin-like and trypsin-like enzyme activity were detected in all gut fluids and tissues tested, with chymotrypsin displaying the highest activity in saliva and salivary gland tissue, whereas maximal trypsin activity was evident in the crop. Pepsin-like activity was only evident in crop fluids and tissues. The activity of all three enzymes was low or undetectable (pepsin) in the fluids and tissue homogenates derived from the esophagus and cardia of any of the adults assayed. Fed adult females displayed higher enzyme activities than fed males, and the activity of all three enzymes were much more prevalent in fed adults than starved. The pH optimum of the trypsin-like enzyme was between pH 7.0 and 8.0; chymotrypsin was near pH 8.0; and maximal pepsin-like activity occurred between pH 1.0 and 2.0. Regurgitate from fed adult females displayed enzyme activity consistent with the proteolytic enzymes detected in crop gut fluids. Enzymes in regurgitate were not derived from food sources based on assays of bovine liver samples. These latter observations suggest that adult flies release fluids from foregut when encountering dry foods, potentially as a means to initiate extra-oral digestion. Copyright © 2014 Elsevier Ltd. All rights reserved.

Lead action on activity of some enzymes of plants

International Nuclear Information System (INIS)

Korolyov, A.N.; Koshkaryova, A.I.

2008-01-01

Lead action on activity of some enzymes of young plants of barley double-row (Hordeum distichon L.) families of cereals (Grominea). It is established that activity urease, catalase, ascorbatoxidase is in dependence as from a lead dose in a nutritious solution, and term ontogenesis. At later stages ontogenesis the increase in concentration of lead in an inhabitancy leads to sharp decrease in activity ascorbatoxidase. In the same conditions activity urease and catalase raises.

Serial crystallography captures enzyme catalysis in copper nitrite reductase at atomic resolution from one crystal

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Sam Horrell

2016-07-01

Full Text Available Relating individual protein crystal structures to an enzyme mechanism remains a major and challenging goal for structural biology. Serial crystallography using multiple crystals has recently been reported in both synchrotron-radiation and X-ray free-electron laser experiments. In this work, serial crystallography was used to obtain multiple structures serially from one crystal (MSOX to study in crystallo enzyme catalysis. Rapid, shutterless X-ray detector technology on a synchrotron MX beamline was exploited to perform low-dose serial crystallography on a single copper nitrite reductase crystal, which survived long enough for 45 consecutive 100 K X-ray structures to be collected at 1.07–1.62 Å resolution, all sampled from the same crystal volume. This serial crystallography approach revealed the gradual conversion of the substrate bound at the catalytic type 2 Cu centre from nitrite to nitric oxide, following reduction of the type 1 Cu electron-transfer centre by X-ray-generated solvated electrons. Significant, well defined structural rearrangements in the active site are evident in the series as the enzyme moves through its catalytic cycle, namely nitrite reduction, which is a vital step in the global denitrification process. It is proposed that such a serial crystallography approach is widely applicable for studying any redox or electron-driven enzyme reactions from a single protein crystal. It can provide a `catalytic reaction movie' highlighting the structural changes that occur during enzyme catalysis. The anticipated developments in the automation of data analysis and modelling are likely to allow seamless and near-real-time analysis of such data on-site at some of the powerful synchrotron crystallographic beamlines.

Activity of an enzyme immobilized on superparamagnetic particles in a rotational magnetic field

Energy Technology Data Exchange (ETDEWEB)

Mizuki, Toru; Watanabe, Noriyuki; Nagaoka, Yutaka [Bio-Nano Electronics Research Centre, Toyo University, Saitama 350-8585 (Japan); Fukushima, Tadamasa [Shimadzu GLC Ltd., Phenomenex Support Centre, Tokyo 110-0016 (Japan); Morimoto, Hisao; Usami, Ron [Bio-Nano Electronics Research Centre, Toyo University, Saitama 350-8585 (Japan); Maekawa, Toru, E-mail: maekawa@toyonet.toyo.ac.jp [Bio-Nano Electronics Research Centre, Toyo University, Saitama 350-8585 (Japan)

2010-03-19

We immobilize {alpha}-amylase extracted from Bacillus Iicheniformis on the surfaces of superparamagnetic particles and investigate the effect of a rotational magnetic field on the enzyme's activity. We find that the activity of the enzyme molecules immobilized on superparamagnetic particles increases in the rotational magnetic field and reaches maximum at a certain frequency. We clarify the effect of the cluster structures formed by the superparamagnetic particles on the activity. Enzyme reactions are enhanced even in a tiny volume of solution using the present method, which is very important for the development of efficient micro reactors and micro total analysis systems ({mu}-TAS).

In vivo enzyme activity in inborn errors of metabolism

Energy Technology Data Exchange (ETDEWEB)

Thompson, G.N.; Walter, J.H.; Leonard, J.V.; Halliday, D. (Clinical Research Centre, Harrow (England))

1990-08-01

Low-dose continuous infusions of (2H5)phenylalanine, (1-13C)propionate, and (1-13C)leucine were used to quantitate phenylalanine hydroxylation in phenylketonuria (PKU, four subjects), propionate oxidation in methylmalonic acidaemia (MMA, four subjects), and propionic acidaemia (PA, four subjects) and leucine oxidation in maple syrup urine disease (MSUD, four subjects). In vivo enzyme activity in PKU, MMA, and PA subjects was similar to or in excess of that in adult controls (range of phenylalanine hydroxylation in PKU, 3.7 to 6.5 mumol/kg/h, control 3.2 to 7.9, n = 7; propionate oxidation in MMA, 15.2 to 64.8 mumol/kg/h, and in PA, 11.1 to 36.0, control 5.1 to 19.0, n = 5). By contrast, in vivo leucine oxidation was undetectable in three of the four MSUD subjects (less than 0.5 mumol/kg/h) and negligible in the remaining subject (2 mumol/kg/h, control 10.4 to 15.7, n = 6). These results suggest that significant substrate removal can be achieved in some inborn metabolic errors either through stimulation of residual enzyme activity in defective enzyme systems or by activation of alternate metabolic pathways. Both possibilities almost certainly depend on gross elevation of substrate concentrations. By contrast, only minimal in vivo oxidation of leucine appears possible in MSUD.

In vivo enzyme activity in inborn errors of metabolism

International Nuclear Information System (INIS)

Thompson, G.N.; Walter, J.H.; Leonard, J.V.; Halliday, D.

1990-01-01

Low-dose continuous infusions of [2H5]phenylalanine, [1-13C]propionate, and [1-13C]leucine were used to quantitate phenylalanine hydroxylation in phenylketonuria (PKU, four subjects), propionate oxidation in methylmalonic acidaemia (MMA, four subjects), and propionic acidaemia (PA, four subjects) and leucine oxidation in maple syrup urine disease (MSUD, four subjects). In vivo enzyme activity in PKU, MMA, and PA subjects was similar to or in excess of that in adult controls (range of phenylalanine hydroxylation in PKU, 3.7 to 6.5 mumol/kg/h, control 3.2 to 7.9, n = 7; propionate oxidation in MMA, 15.2 to 64.8 mumol/kg/h, and in PA, 11.1 to 36.0, control 5.1 to 19.0, n = 5). By contrast, in vivo leucine oxidation was undetectable in three of the four MSUD subjects (less than 0.5 mumol/kg/h) and negligible in the remaining subject (2 mumol/kg/h, control 10.4 to 15.7, n = 6). These results suggest that significant substrate removal can be achieved in some inborn metabolic errors either through stimulation of residual enzyme activity in defective enzyme systems or by activation of alternate metabolic pathways. Both possibilities almost certainly depend on gross elevation of substrate concentrations. By contrast, only minimal in vivo oxidation of leucine appears possible in MSUD

Correlation of denitrification-accepted fraction of electrons with NAD(P)H fluorescence for Pseudomonas aeruginosa performing simultaneous denitrification and respiration at extremely low dissolved oxygen conditions.

Science.gov (United States)

Chen, Fan; Xia, Qing; Ju, Lu-Kwang

2004-01-01

In cystic fibrosis airway infection, Pseudomonas aeruginosa forms a microaerobic biofilm and undergoes significant physiological changes. It is important to understand the bacterium's metabolism at microaerobic conditions. In this work, the culture properties and two indicators (the denitrification-accepted e- fraction and an NAD(P)H fluorescence fraction) for the culture's "fractional approach" to a fully anaerobic denitrifying state were examined in continuous cultures with practically zero DO but different aeration rates. With decreasing aeration, specific OUR decreased while specific NAR and NIR increased and kept Y(ATP/S) relatively constant. P. aeruginosa thus appeared to effectively compensate for energy generation at microaerobic conditions with denitrification. At the studied dilution rate of 0.06 h(-1), the maximum specific OUR was 2.8 mmol O2/g cells-h and the Monod constant for DO, in the presence of nitrate, was extremely low (Y(X/S) increased significantly (from 0.24 to 0.34) with increasing aeration, attributed to a roughly opposite trend of Y(ATP/X) (ATP generation required for cell growth). As for the denitrification-accepted e- fraction and the fluorescence fraction, both decreased with increasing aeration as expected. The two fractions, however, were not directly proportional. The fluorescence fraction changed more rapidly than the e- fraction at very low aeration rates, whereas the opposite was true at higher aeration. The results demonstrated the feasibility of using online NAD(P)H fluorescence to monitor sensitive changes of cellular physiology and provided insights to the shift of e- -accepting mechanisms of P. aeruginosa under microaerobic conditions.

Spatial variability in denitrification rates in an Oregon tidal salt marsh

Science.gov (United States)

Modeling denitrification (DeN) is particularly challenging in tidal systems, which play a vital role in buffering adjacent coastal waters from nitrogen inputs. These systems are hydrologically and biogeochemically complex, varying on fine temporal and spatial scales. As part of a...

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

African Journals Online (AJOL)

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

First glycoside hydrolase family 2 enzymes from Thermus antranikianii and Thermus brockianus with β-glucosidase activity

Directory of Open Access Journals (Sweden)

Carola eSchröder

2015-06-01

Full Text Available Two genes tagh2 and tbgh2 coding for enzymes with hydrolytic activity towards esculin were identified from the extreme thermophilic, aerobic bacteria Thermus antranikianii (Ta and T. brockianus (Tb. Shortened conserved domains predicted a membership of the enzymes of glycoside hydrolase (GH family 2. At present, β-galactosidase activity is found frequently in GH family 2 but β-glucosidase activity has not been reported in this family before. The enzymes TaGH2 and TbGH2 preferred hydrolysis of nitrophenol-linked β-D-glucopyranosides with specific activities of 3,966 U/mg and 660 U/mg, respectively. Residual activities of 40 % (TaGH2 and 51 % (TbGH2 towards 4-NP-β-D-galactopyranoside were observed. Furthermore, TaGH2 hydrolyzed cellobiose. TbGH2, however, showed no activity on cellobiose or lactose. The enzymes exhibited highest activity at 95 °C (TaGH2 and 90 °C (TbGH2 at pH 6.5. Both enzymes were extremely thermostable and thermal activation up to 250 % was observed at temperatures between 50 and 60 °C. Accordingly, the first thermoactive glycoside hydrolase family 2 enzymes with β glucosidase activity have been identified and characterized. The hydrolysis of cellobiose is a unique property of TaGH2 when compared to the enzymes of GH family 2.

activity of enzyme trypsin immobilized onto macroporous poly(epoxy

African Journals Online (AJOL)

dell

consequential effects of covalent immobilization. EXPERIMENTAL. Materials .... immersed into water bath. ... storage stability of the enzyme was studied ... pore size range of about 10 to 150 µm. ... figures, the differences in activities (slopes.

Changes in photosynthesis and activities of enzymes involved in ...

African Journals Online (AJOL)

tolerance, respectively were used to investigate the oxygen consumption rate of photosystem I, the oxygen evolution rate of photosystem II, cab transcript levels, and activities of enzymes involved in photosynthetic carbon reduction cycle.

Enzyme-activity mutations detected in mice after paternal fractionated irradiation

International Nuclear Information System (INIS)

Charles, D.J.; Pretsch, W.

1986-01-01

(101/E1 X C3H/E1)F 1 -hybrid male mice were exposed in a 24-h fractionation interval to either 3.0 + 3.0-Gy or 5.1 + 5.1-Gy X-irradiation, and mated to untreated Test-stock females. The offspring were examined for mutations at 7 recessive specific loci and for activity alterations of erythrocyte enzymes controlled presumably by 12 loci. No enzyme-activity mutant was found in 3610 F 1 -offspring of the control group. In the experimental groups, no mutant was detected in 533 (3.0 + 3.0 Gy) and 173 (5.1 + 5.1 Gy) offspring from postspermatogonial germ cells treated. After treatment of spermatogonia, 1 mutant in 3388 F 1 -offspring of the 3.0 + 3.0-Gy group, and 5 mutants in 3187 F 1 offspring of the 5.1 + 5.1-Gy group were found. The mutants were all genetically confirmed. The frequency (expressed as mutants/locus/gamete) of enzyme-activity mutations is 2 (5.1 + 5.1-Gy group) to 10 (3.0 + 3.0-Gy group) times lower than the frequency of recessive specific-locus mutations. (Auth.)

Embryonic turkey liver: activities of biotransformation enzymes and activation of DNA-reactive carcinogens

International Nuclear Information System (INIS)

Perrone, Carmen E.; Duan, Jian Dong; Jeffrey, Alan M.; Williams, Gary M.; Ahr, Hans-Juergen; Schmidt, Ulrich; Enzmann, Harald H.

2004-01-01

Avian embryos are a potential alternative model for chemical toxicity and carcinogenicity research. Because the toxic and carcinogenic effects of some chemicals depend on bioactivation, activities of biotransformation enzymes and formation of DNA adducts in embryonic turkey liver were examined. Biochemical analyses of 22-day in ovoturkey liver post-mitochondrial fractions revealed activities of the biotransformation enzymes 7-ethoxycoumarin de-ethylase (ECOD), 7-ethoxyresorufin de-ethylase (EROD), aldrin epoxidase (ALD), epoxide hydrolase (EH), glutathione S-transferase (GST), and UDP-glucuronyltransferase (GLUT). Following the administration of phenobarbital (24 mg/egg) on day 21, enzyme activities of ECOD, EROD, ALD, EH and GLUT, but not of GST, were increased by two-fold or higher levels by day 22. In contrast, acute administration of 3-methylcholanthrene (5 mg/egg) induced only ECOD and EROD activities. Bioactivation of structurally diverse pro-carcinogens was also examined using 32 P-postlabeling for DNA adducts. In ovoexposure of turkey embryos on day 20 of gestation to 2-acetylaminofluorene (AAF), 4,4'-methylenebis(2-chloroaniline) (MOCA), benzo[a]pyrene (BaP), and 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) resulted in the formation of DNA adducts in livers collected by day 21. Some of the DNA adducts had 32 P-postlabeling chromatographic migration patterns similar to DNA adducts found in livers from Fischer F344 rats exposed to the same pro-carcinogens. We conclude that 21-day embryonic turkey liver is capable of chemical biotransformation and activation of genotoxic carcinogens to form DNA adducts. Thus, turkey embryos could be utilized to investigate potential chemical toxicity and carcinogenicity. (orig.)

Embryonic turkey liver: activities of biotransformation enzymes and activation of DNA-reactive carcinogens

Energy Technology Data Exchange (ETDEWEB)

Perrone, Carmen E.; Duan, Jian Dong; Jeffrey, Alan M.; Williams, Gary M. [New York Medical College, Department of Pathology, Valhalla (United States); Ahr, Hans-Juergen; Schmidt, Ulrich [Bayer AG, Institute of Toxicology, Wuppertal (Germany); Enzmann, Harald H. [Federal Institute for Drugs and Medical Devices, Bonn (Germany)

2004-10-01

Avian embryos are a potential alternative model for chemical toxicity and carcinogenicity research. Because the toxic and carcinogenic effects of some chemicals depend on bioactivation, activities of biotransformation enzymes and formation of DNA adducts in embryonic turkey liver were examined. Biochemical analyses of 22-day in ovoturkey liver post-mitochondrial fractions revealed activities of the biotransformation enzymes 7-ethoxycoumarin de-ethylase (ECOD), 7-ethoxyresorufin de-ethylase (EROD), aldrin epoxidase (ALD), epoxide hydrolase (EH), glutathione S-transferase (GST), and UDP-glucuronyltransferase (GLUT). Following the administration of phenobarbital (24 mg/egg) on day 21, enzyme activities of ECOD, EROD, ALD, EH and GLUT, but not of GST, were increased by two-fold or higher levels by day 22. In contrast, acute administration of 3-methylcholanthrene (5 mg/egg) induced only ECOD and EROD activities. Bioactivation of structurally diverse pro-carcinogens was also examined using {sup 32}P-postlabeling for DNA adducts. In ovoexposure of turkey embryos on day 20 of gestation to 2-acetylaminofluorene (AAF), 4,4'-methylenebis(2-chloroaniline) (MOCA), benzo[a]pyrene (BaP), and 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) resulted in the formation of DNA adducts in livers collected by day 21. Some of the DNA adducts had {sup 32}P-postlabeling chromatographic migration patterns similar to DNA adducts found in livers from Fischer F344 rats exposed to the same pro-carcinogens. We conclude that 21-day embryonic turkey liver is capable of chemical biotransformation and activation of genotoxic carcinogens to form DNA adducts. Thus, turkey embryos could be utilized to investigate potential chemical toxicity and carcinogenicity. (orig.)

The effect of millennial-scale changes in Arabian Sea denitrification on atmospheric CO{sub 2}

Energy Technology Data Exchange (ETDEWEB)

Altabet, M.A.; Higginson, M.J. [University of Massachusetts, New Bedford, MA (United States). School for Marine Science and Technology; Murray, D.W. [Brown University, Providence, RI (United States). Center for Environmental Studies

2002-07-01

Most global biogeochemical processes are known to respond to climate change, some of which have the capacity to produce feedbacks through the regulation of atmospheric greenhouse gases. Marine denitrification - the reduction of nitrate to gaseous nitrogen - is an important process in this regard, affecting greenhouse gas concentrations directly through the incidental production of nitrous oxide, and indirectly through modification of the marine nitrogen inventory and hence the biological pump for C0{sub 2}. Although denitrification has been shown to vary with glacial-interglacial cycles, its response to more rapid climate change has not yet been well characterized. Here we present nitrogen isotope ratio, nitrogen content and chlorin abundance data from sediment cores with high accumulation rates on the Oman continental margin that reveal substantial millennial-scale variability in Arabian Sea denitrification and productivity during the last glacial period. The detailed correspondence of these changes with Dansgaard-Oeschger events recorded in Greenland ice cores indicates rapid, century-scale reorganization of the Arabian Sea ecosystem in response to climate excursions, mediated through the intensity of summer monsoonal upwelling. Considering the several-thousand-year residence time of fixed nitrogen in the ocean, the response of global marine productivity to changes in denitrification would have occurred at lower frequency and appears to be related to climatic and atmospheric C0{sub 2} oscillations observed in Antarctic ice cores between 20 and A kyr ago. (author)

Probiotic activity of lignocellulosic enzyme as bioactivator for rice husk degradation

Science.gov (United States)

Lamid, Mirni; Al-Arif, Anam; Warsito, Sunaryo Hadi

2017-02-01

The utilization of lignocellulosic enzyme will increase nutritional value of rice husk. Cellulase consists of C1 (β-1, 4-glucan cellobiohydrolase or exo-β-1,4glucanase), Cc (endo-β-1,4-glucanase) and component and cellobiose (β-glucocidase). Hemicellulase enzyme consists of endo-β-1,4-xilanase, β-xilosidase, α-L arabinofuranosidase, α-D-glukuronidaseand asetil xilan esterase. This research aimed to study the activity of lignocellulosic enzyme, produced by cows in their rumen, which can be used as a bioactivator in rice husk degradation. This research resulted G6 and G7 bacteria, producing xylanase and cellulase with the activity of 0.004 U mL-1 and 0.021 U mL-1; 0.003 ( U mL-1) and 0.026 (U mL-1) respectively.

Nitrifier-induced denitrification is an important source of soil nitrous oxide and can be inhibited by a nitrification inhibitor 3,4-dimethylpyrazole phosphate.

Science.gov (United States)

Shi, Xiuzhen; Hu, Hang-Wei; Zhu-Barker, Xia; Hayden, Helen; Wang, Juntao; Suter, Helen; Chen, Deli; He, Ji-Zheng

2017-12-01

Soil ecosystem represents the largest contributor to global nitrous oxide (N 2 O) production, which is regulated by a wide variety of microbial communities in multiple biological pathways. A mechanistic understanding of these N 2 O production biological pathways in complex soil environment is essential for improving model performance and developing innovative mitigation strategies. Here, combined approaches of the 15 N- 18 O labelling technique, transcriptome analysis, and Illumina MiSeq sequencing were used to identify the relative contributions of four N 2 O pathways including nitrification, nitrifier-induced denitrification (nitrifier denitrification and nitrification-coupled denitrification) and heterotrophic denitrification in six soils (alkaline vs. acid soils). In alkaline soils, nitrification and nitrifier-induced denitrification were the dominant pathways of N 2 O production, and application of the nitrification inhibitor 3,4-dimethylpyrazole phosphate (DMPP) significantly reduced the N 2 O production from these pathways; this is probably due to the observed reduction in the expression of the amoA gene in ammonia-oxidizing bacteria (AOB) in the DMPP-amended treatments. In acid soils, however, heterotrophic denitrification was the main source for N 2 O production, and was not impacted by the application of DMPP. Our results provide robust evidence that the nitrification inhibitor DMPP can inhibit the N 2 O production from nitrifier-induced denitrification, a potential significant source of N 2 O production in agricultural soils. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

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

Science.gov (United States)

Mganga, Kevin; Razavi, Bahar; Kuzyakov, Yakov

2015-04-01

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

A comparison of maximal bioenergetic enzyme activities obtained with commonly used homogenization techniques.

Science.gov (United States)

Grace, M; Fletcher, L; Powers, S K; Hughes, M; Coombes, J

1996-12-01

Homogenization of tissue for analysis of bioenergetic enzyme activities is a common practice in studies examining metabolic properties of skeletal muscle adaptation to disease, aging, inactivity or exercise. While numerous homogenization techniques are in use today, limited information exists concerning the efficacy of specific homogenization protocols. Therefore, the purpose of this study was to compare the efficacy of four commonly used approaches to homogenizing skeletal muscle for analysis of bioenergetic enzyme activity. The maximal enzyme activity (Vmax) of citrate synthase (CS) and lactate dehydrogenase (LDH) were measured from homogenous muscle samples (N = 48 per homogenization technique) and used as indicators to determine which protocol had the highest efficacy. The homogenization techniques were: (1) glass-on-glass pestle; (2) a combination of a mechanical blender and a teflon pestle (Potter-Elvehjem); (3) a combination of the mechanical blender and a biological detergent; and (4) the combined use of a mechanical blender and a sonicator. The glass-on-glass pestle homogenization protocol produced significantly higher (P pestle homogenization protocol is the technique of choice for studying bioenergetic enzyme activity in skeletal muscle.

Co-ordinate activation of lipogenic enzymes in hepatocellular carcinoma.

Science.gov (United States)

Yahagi, Naoya; Shimano, Hitoshi; Hasegawa, Kiyoshi; Ohashi, Kenichi; Matsuzaka, Takashi; Najima, Yuho; Sekiya, Motohiro; Tomita, Sachiko; Okazaki, Hiroaki; Tamura, Yoshiaki; Iizuka, Yoko; Ohashi, Ken; Nagai, Ryozo; Ishibashi, Shun; Kadowaki, Takashi; Makuuchi, Masatoshi; Ohnishi, Shin; Osuga, Jun-ichi; Yamada, Nobuhiro

2005-06-01

Hepatocellular carcinoma is a very common neoplastic disease in countries where hepatitis viruses B and/or C are prevalent. Small hepatocellular carcinoma lesions detected by ultrasonography at an early stage are often hyperechoic because they are composed of well-differentiated cancer cells that are rich in triglyceride droplets. The triglyceride content of hepatocytes depends in part on the rate of lipogenesis. Key lipogenic enzymes, such as fatty acid synthase, are co-ordinately regulated at the transcriptional level. We therefore examined the mRNA expression of lipogenic enzymes in human hepatocellular carcinoma samples from 10 patients who had undergone surgical resection. All of the samples exhibited marked elevation of expression of mRNA for lipogenic enzymes, such as fatty acid synthase, acetyl-CoA carboxylase and ATP citrate lyase, compared with surrounding non-cancerous liver tissue. In contrast, the changes in mRNA expression of SREBP-1, a transcription factor that regulates a battery of lipogenic enzymes, did not show a consistent trend. In some cases where SREBP-1 was elevated, the main contributing isoform was SREBP-1c rather than SREBP-1a. Thus, lipogenic enzymes are markedly induced in hepatocellular carcinomas, and in some cases SREBP-1c is involved in this activation.

Effects of electrokinetic treatment of a heavy metal contaminated soil on soil enzyme activities

International Nuclear Information System (INIS)

Cang Long; Zhou Dongmei; Wang Quanying; Wu Danya

2009-01-01

There is a growing concern on the potential application of a direct current (DC) electric field to soil for removing contaminants, but little is known about its impact on soil enzyme activities. This study investigated the change of enzyme activities of a heavy metal contaminated soil before and after electrokinetic (EK) treatments at lab-scale and the mechanisms of EK treatment to affect soil enzyme activities were explored. After treatments with 1-3 V cm -1 of voltage gradient for 420 h, soil pH, electrical conductivity (EC), soil organic carbon, dissolved organic carbon (DOC), soil heavy metal concentration and enzyme activities were analyzed. The results showed that the average removal efficiencies of soil copper were about 65% and 83% without and with pH control of catholyte, respectively, and all the removal efficiencies of cadmium were above 90%. The soil invertase and catalase activities increased and the highest invertase activity was as 170 times as the initial one. The activities of soil urease and acidic phosphatase were lower than the initial ones. Bivariate correlation analyses indicated that the soil invertase and acidic phosphatase activities were significantly correlated with soil pH, EC, and DOC at P < 0.05, but the soil urease activities had no correlation with the soil properties. On the other hand, the effects of DC electric current on solution invertase and catalase enzyme protein activities indicated that it had negative effect on solution catalase activity and little effect on solution invertase activity. From the change of invertase and catalase activities in soil and solution, the conclusion can be drawn that the dominant effect mechanism is the change of soil properties by EK treatments.

Effects of electrokinetic treatment of a heavy metal contaminated soil on soil enzyme activities

Energy Technology Data Exchange (ETDEWEB)

Cang Long [State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences, Nanjing 210008 (China); Graduate School of the Chinese Academy of Sciences, Beijing 100049 (China); Zhou Dongmei, E-mail: dmzhou@issas.ac.cn [State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences, Nanjing 210008 (China); Wang Quanying; Wu Danya [State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences, Nanjing 210008 (China); Graduate School of the Chinese Academy of Sciences, Beijing 100049 (China)

2009-12-30

There is a growing concern on the potential application of a direct current (DC) electric field to soil for removing contaminants, but little is known about its impact on soil enzyme activities. This study investigated the change of enzyme activities of a heavy metal contaminated soil before and after electrokinetic (EK) treatments at lab-scale and the mechanisms of EK treatment to affect soil enzyme activities were explored. After treatments with 1-3 V cm{sup -1} of voltage gradient for 420 h, soil pH, electrical conductivity (EC), soil organic carbon, dissolved organic carbon (DOC), soil heavy metal concentration and enzyme activities were analyzed. The results showed that the average removal efficiencies of soil copper were about 65% and 83% without and with pH control of catholyte, respectively, and all the removal efficiencies of cadmium were above 90%. The soil invertase and catalase activities increased and the highest invertase activity was as 170 times as the initial one. The activities of soil urease and acidic phosphatase were lower than the initial ones. Bivariate correlation analyses indicated that the soil invertase and acidic phosphatase activities were significantly correlated with soil pH, EC, and DOC at P < 0.05, but the soil urease activities had no correlation with the soil properties. On the other hand, the effects of DC electric current on solution invertase and catalase enzyme protein activities indicated that it had negative effect on solution catalase activity and little effect on solution invertase activity. From the change of invertase and catalase activities in soil and solution, the conclusion can be drawn that the dominant effect mechanism is the change of soil properties by EK treatments.