Absence Of External Electron Donor Research Articles

Modelling bioelectrochemical denitrification in absence of electron donors for groundwater treatment

Microbial electrochemical technologies (METs) have become a widely studied technology in recent years due to the need for sustainable biotechnologies. The scope of this work is the development of a mechanistic biokinetic model, based on first principles and a robust thermodynamic basis, to provide a theoretical accurate description of a MET system that would treat water contaminated with nitrate, the most common aquifer water pollutant, in absence of external electron donors. The model aims at describing the complex processes occurring including the competition between bioelectroactive and non-bioelectroactive reactions as well as the dynamics and kinetics of multiple bioelectrochemical reactions (both in series and in parallel) taking place in the same electrode. The bioelectrochemical denitrification of groundwater was then evaluated using the model as a case study. The evaluation focused on theoretical removal rates and energy expenditure, as well as the effect of key design parameters on the system's performance. The model successfully described how changes in the applied voltage and/or hydraulic retention time may impact the performance in terms of removal rate and effluent quality. The theoretical results also predict that the impact of electrode area is potentially more significant on the energy efficiency rather than on the effluent quality.

How hydrogen peroxide is metabolized by oxidized cytochrome c oxidase.

In the absence of external electron donors, oxidized bovine cytochrome c oxidase (CcO) exhibits the ability to decompose excess H2O2. Depending on the concentration of peroxide, two mechanisms of degradation were identified. At submillimolar peroxide concentrations, decomposition proceeds with virtually no production of superoxide and oxygen. In contrast, in the millimolar H2O2 concentration range, CcO generates superoxide from peroxide. At submillimolar concentrations, the decomposition of H2O2 occurs at least at two sites. One is the catalytic heme a3–CuB center where H2O2 is reduced to water. During the interaction of the enzyme with H2O2, this center cycles back to oxidized CcO via the intermediate presence of two oxoferryl states. We show that at pH 8.0 two molecules of H2O2 react with the catalytic center accomplishing one cycle. In addition, the reactions at the heme a3–CuB center generate the surface-exposed lipid-based radical(s) that participates in the decomposition of peroxide. It is also found that the irreversible decline of the catalytic activity of the enzyme treated with submillimolar H2O2 concentrations results specifically from the decrease in the rate of electron transfer from heme a to the heme a3–CuB center during the reductive phase of the catalytic cycle. The rates of electron transfer from ferrocytochrome c to heme a and the kinetics of the oxidation of the fully reduced CcO with O2 were not affected in the peroxide-modified CcO.

Permeable reactive biobarriers for in situ Cr(VI) reduction: Bench scale tests using Cellulomonas sp. strain ES6

Chromate (Cr(VI)) reduction studies were performed in bench scale flow columns using the fermentative subsurface isolate Cellulomonas sp. strain ES6. In these tests, columns packed with either quartz sand or hydrous ferric oxide (HFO)-coated quartz sand, were inoculated with strain ES6 and fed nutrients to stimulate growth before nutrient-free Cr(VI) solutions were injected. Results show that in columns containing quartz sand, a continuous inflow of 2 mg/L Cr(VI) was reduced to below detection limits in the effluent for durations of up to 5.7 residence times after nutrient injection was discontinued proving the ability of strain ES6 to reduce chromate in the absence of an external electron donor. In the HFO-containing columns, Cr(VI) reduction was significantly prolonged and effluent Cr(VI) concentrations remained below detectable levels for periods of up to 66 residence times after nutrient injection was discontinued. Fe was detected in the effluent of the HFO-containing columns throughout the period of Cr(VI) removal indicating that the insoluble Fe(III) bearing solids were being continuously reduced to form soluble Fe(II) resulting in prolonged abiotic Cr(VI) reduction. Thus, growth of Cellulomonas within the soil columns resulted in formation of permeable reactive barriers that could reduce Cr(VI) and Fe(III) for extended periods even in the absence of external electron donors. Other bioremediation systems employing Fe(II)-mediated reactions require a continuous presence of external nutrients to regenerate Fe(II). After depletion of nutrients, contaminant removal within these systems occurs by reaction with surface-associated Fe(II) that can rapidly become inaccessible due to formation of crystalline Fe-minerals or other precipitates. The ability of fermentative organisms like Cellulomonas to reduce metals without continuous nutrient supply in the subsurface offers a viable and economical alternative technology for in situ remediation of Cr(VI)-contaminated groundwater through formation of permeable reactive biobarriers (PRBB).

Microbial Selenate Sorption and Reduction in Nutrient Limited Systems

In this study, batch sorption experiments and X-ray adsorption spectroscopy (XAS) were utilized to investigate selenate sorption onto Shewanella putrefaciens 200R. Selenate sorption was studied as a function of pH (ranging from 3 to 7), ionic strength (ranging from 0.1 to 0.001 M), and initial selenate concentration (ranging from 10 to 5000 microM) in the absence of external electron donors. The results show that the extent of selenate sorption is strongly dependent on pH and ionic strength, with maximum sorption occurring at low pH (pH = 3) and low ionic strength (0.001 M NaCl) conditions. The strong dependence of Se sorption with ionic strength suggests the formation of outersphere complexes with the cell wall functional groups. Langmuir isotherm plots yielded log Kads values from 2.74 to 3.02. Desorption experiments demonstrated thatthe binding of selenate onto S. putrefaciens was not completely reversible. XANES analysis of the cells after sorption experiments revealed the presence of elemental selenium, indicating that S. putrefaciens has a capacity to reduce Se(VI) to Se(0) in the absence of external electron donors. We conclude that Se sorption onto S. putrefaciens cell walls is the result of the combination of outer-sphere complexation and cell surface reduction. This sorption process leads to a complex reservoir of bound Se which is not entirely reversible.

Long-term tetrachlorethene degradation sustained by endogenous cell decay

The role of endogenous decay in the reductive dechlorination of tetrachlorethene (PCE) was studied in a sand column containing an anaerobic microbial consortium that included Dehalococcoides ethenogenes. The column consortium had continuously transformed PCE to vinyl chloride (<5%) and ethene (>95%) with the addition of ethanol and yeast extract for a 3 year period. The column was operated for a further period of 1 year with the addition of 60 µM PCE only. In the absence of external electron donors, PCE dechlorination in the column initially slowed but continued at a relatively constant rate for 1 year. The conversion to vinyl chloride (VC) and ethene (ETH) was significantly reduced with cis-dichloroethylene (cis-DCE) becoming the major (90%) end product of dechlorination. Microcosm studies, with no external electron donor added, indicated that reductive dechlorination was sustained by reducing equivalents derived from endogenous decay products. Measures of cellular adenosine triphosphate (ATP) along the length of the column indicated that even after 1 year of only PCE addition, an active biological community was sustained. Key words: tetrachloroethene (PCE), ethanol, anaerobic biodegradation, endogenous decay, column study.

Biodegradation of 1,1,1,2-tetrachloroethane under methanogenic conditions

Chlorinated aliphatic hydrocarbons are widely used as solvents and as intermediates in chemical synthesis, so they can be found in industrial wastewaters and released to the environment where they became a serious health risk due to their toxic properties and high chemical stability. Most of these compounds are xenobiotic and recalcitrant to biodegradation. In this article we report the effect of different co-substrates in the 1,1,1,2-tetrachloroethane (1,1,1,2-TeCA) degradation by anaerobic granular sludge, and its degradative pathway. Our results show that this compound is easy and rapidly biodegradable under methanogenic conditions, even in the absence of external electron donors. 1,1,1,2-TeCA is equimolecularly degraded to 1,1-dichloroethene (1,1-DCE) by reductive dichloroelimination. 1,1-DCE is only completely biodegraded in the presence of lactic acid as co-substrate. Although 1,1,1,2-TeCA can be apparently removed by autoclaved granular sludge, the compound is not transformed but retained inside the granules. The primary biodegradation of 1,1,1,2-TeCE to 1,1-DCE is a biotic process mediated by anaerobic bacteria.

Methane microprofiles in a sewage biofilm determined with a microscale biosensor

Microprofiles of the methane concentration in a 3.5-mm-thick sewage outlet biofilm were measured at high spatial and temporal resolution using a microscale biosensor for methane. In the freshly collected biofilm, methane was building up to a concentration of 175 μmol l −1 at 3 mm depth with a total methanogenesis of 0.14 μmol m −2 s −1, as compared to an aerobic respiration (including methane oxidation) of 0.80 μmol m −2 s −1. A model biofilm was established by homogenisation of an in situ biofilm and 12 days of incubation with surplus sodium acetate. The homogenised biofilm was able to maintain 50% of the methanogenic activity in the absence of external electron donor. Oxygen had only a minor effect on the methane production, but aerobic respiration consumed a substantial part of the produced methane and was thus an important control on methane export from the biofilm. A concentration of 2 mmol l −1 nitrate was shown to inhibit methanogenesis only in the upper layer of the biofilm, whereas a further addition of 2 mmo1 l −1 sulphate inhibited methanogenesis in the entire biofilm. The study demonstrated the power of the methane microsensor in the study of microhabitats with concurrent production and consumption of methane.

Inter-flavin electron transfer in the ground state in the absence of external electron donors and acceptors

Inter-flavin electron transfer of N5-alkylflavinium cation 3 occurred in the ground state to primarily give the flavosemiquinone 4 and the superoxidized flavin 7. This provided insight into some chemistry of 7: a rapid addition followed by a (second) inter-flavin electron transfer of the derived 4 a- flavin adduct radical 6 giving 3 and, consequently, an additional formation of 4.

X-ray Absorption and EPR Studies on the Copper Ions Associated with the Particulate Methane Monooxygenase from Methylococcus capsulatus (Bath). Cu(I) Ions and Their Implications

Parallel X-ray absorption edge and EPR studies of the particulate methane monooxygenase in situ reveal that the enzyme contains unusually high levels of copper ions with a significant portion of the copper ions existing as Cu(I) in the “as-isolated” form (70−80%). The observation of high levels of reduced copper in a monooxygenase is surprising considering that the natural cosubstrate of the enzyme is dioxygen. Toward clarifying the roles of the various copper ions in the enzyme, we have successfully prepared different states of the protein in the membrane-bound form at various levels of reduction using dithionite, dioxygen, and ferricyanide. EPR intensity analysis of the fully-oxidized preparations indicates that the bulk of copper ions are arranged in cluster units. The fully-reduced protein obtained by reduction by dithionite has been used to initiate the single turnover of the enzyme in the presence of dioxygen. Differential reactivity toward dioxygen was observed upon analyzing the copper reduction levels in these synchronized preparations. The enzyme is capable of supporting turnover in the absence of external electron donors in the highly reduced states. These results suggest the presence of at least two classes of copper ions in the particulate methane monooxygenase. As a working hypothesis, we have referred to these classes of copper ions as (1) the catalytic (C) clusters, which function principally as the catalytic core of the enzyme, and (2) the electron-transfer (E) clusters, which are presumed to be the source of endogenous reducing equivalents and therefore function in an electron-transfer capacity.

Pre-steady-state kinetic study on the formation of compound I and II of ligninase.

The reaction between ligninase and hydrogen peroxide yielding Compound I has been investigated using a stopped-flow rapid-scan spectrophotometer. The optical absorption spectrum of Compound I appears different to that reported by Andrawis, A. et al. (1987) and Renganathan, V. and Gold, M.H. (1986), in that the Soret-maximum is at 401 nm rather than 408 nm. The second-order rate constant (4.2.10(5) M-1.s-1) for the formation of Compound I was independent of pH (pH 3.0-6.0). In the absence of external electron donors, Compound I decayed to Compound II with a half-life of 5-10 s at pH 3.1. The rate of this reaction was not affected by the H2O2 concentration used. In the presence of either veratryl alcohol or ferrocyanide, Compound II was rapidly generated. With ferrocyanide, the second-order rate constant increased from 1.9.10(4) M-1.s-1 to 6.8.10(6) M-1.s-1 when the pH was lowered from 6.0 to 3.1. With veratryl alcohol as an electron donor, the second-order rate constant for the formation of Compound II increased from 7.0.10(3) M-1.s-1 at pH 6.0 to 1.0.10(5) M-1.s-1 at pH 4.5. At lower pH values the rate of Compound II formation no longer followed an exponential relationship and the steady-state spectral properties differed to those recorded in the presence of ferrocyanide. Our data support a model of enzyme catalysis in which veratryl alcohol is oxidized in one-electron steps and strengthen the view that veratryl alcohol oxidation involves a substrate-modified Compound II intermediate which is rapidly reduced to the native enzyme.

PHOTOREDUCTION OF NAD TO NADH IN SEMICONDUCTOR DISPERSIONS

Abstract— Band gap illumination of TiO2 (anatase) dispersions in weakly alkaline solutions of nicotinamide‐adenine‐dinucleotide (NAD+) leads in the presence of rhodium trisbipyridyl complex [Rh(bipy)3+3], to continuous generation of biologically active cofactor NADH. Effects of pH, NAD+ and Rh(bipy)3+3 concentration on the efficiency of this photoconversion process are investigated. The reaction proceeds already in aqueous solution in the absence of external electron donors but it is enhanced significantly in the presence of 10% methanol.