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PubMed2005

Reactions of the diiron(IV) intermediate Q in soluble methane monooxygenase with fluoromethanes.

Soluble methane monooxygenases utilize a carboxylate-bridged diiron center and dioxygen to convert methane to methanol. A diiron(IV) oxo intermediate Q is the active species for this process. Alternative substrates and theoretical studies can help elucidate the mechanism. Experimental results for reactions with derivatized methanes were previously modeled by a combination of quantum mechanical/molecular mechanical techniques and the model was extended to predict the relative reactivity of fluoromethane. We therefore studied reactions of Q with CF(n)H(4-n) (n=1-3) to test the prediction. The kinetics of single-turnover reactions of Q with these substrates were monitored by double-mixing stopped-flow optical spectroscopy. For fluoro- and difluoromethane, conversion to the alcohols occurred with second-order rate constants less than that of methane, the values being 28,700 (CH4)>25,000 (CFH3)>9300 (CF2H2) M(-1) s(-1). KIE values for C-H versus C-D activation above the classical limit were observed, requiring modification of the theoretical predictions.

Catalysis↗

PubMed2004

Methane emission from fixed dome biogas plants in hilly and plain regions of northern India.

Methane emissions from the slurry displacement chambers of different fixed dome type biogas plants (Capacity 2 m3) installed in hilly and plain regions of northern India were quantitatively estimated. The slurry temperature in the biogas plants in the hilly areas of Himachal Pradesh at an altitude of 1300 m above mean sea level remains below the lower mesophilic range (16-25 degrees C) during most part of the year. Difference in the ambient temperature under the two climatic conditions of Hills and plain regions affects the CH4 flux. Methane fluxes from the plants ranged between 7 and 120 gm(-2) d(-1). In northern plain, temperature remains warm (21-33 degrees C) throughout the year except during winter (December to January) when there is a steep fall. Seasonal emission of CH4 ranged between 10 and 178 gm(-2) d(-1). The annual average methane emission from the biogas plants in plain areas was 83.1 gm(-2) d(-1) as compared to 43.1 gm(-2) d(-1)in the hilly areas. Annual contribution per plant to the global methane budget from a fixed dome biogas plant (Cap 2 m3) in plain region of northern India was 53.2 kg as compared 22.3 kg in hilly area.

Bioelectric Energy Sources↗

PubMed2005

Hydrolysis rates, methane production and nitrogen solubilisation of grey waste components during anaerobic degradation.

Municipal grey waste (i.e. the remaining fraction in municipal waste management systems in which putrescibles (biowaste) and other recyclables (paper, metals, glass) are source-segregated) was manually sorted into six main fractions on the basis of composition and also separated by sieving (100 mm mesh size) into two fractions, oversized and undersized, respectively. In practice, in waste management plant the oversized fraction is (or will be) used to produce refuse-derived fuel and the undersized landfilled after biological stabilisation. The methane yields and nitrogen solubilisation of the grey waste and the different fractions (all studied samples were first milled to 5 mm particle samples) were determined in a 237-day methane production batch assay and in a water elution test, respectively. The grey waste was found to contained remnants of putrescibles and also a high amount of other biodegradable waste, including packaging, cartons and cardboard, newsprint, textiles and diapers. These waste fractions comprised 41%-w/w of the grey waste and produced 40-210 m3 methane (total solids (TS))(-1) and less than 0.01 g NH4-N kg TS(added)(-1) except diapers which produced 9.8 g NH4-N kg TS(added)(-1) in the batch assays. In the case of the two sieved fractions and on mass bases, most of the methane originated from the oversized fraction, whereas most of the NH4-N was solublised from the undersized fraction. The first-order kinetic model described rather well the degradation of each grey waste fraction and component, showing the different components to be in the range 0.021-0.058 d(-1), which was around one-sixth of the values reported for the source-segregated putrescible fraction of MSW.

Bacteria, Anaerobic↗

PubMed2006

Effect of plant harvest on methane emission from two constructed wetlands designed for the treatment of wastewater.

The emission of methane from two constructed wetlands [a free water surface flow system (FWS) and a subsurface flow system (SF)], constructed for the treatment of waste water, was evaluated at different sites inhabited by reeds (Phragmites communis), to test the effects of plant harvest. High methane emission was recorded immediately after harvesting in both wetlands. Several days after harvesting, the emission decreased in the FWS but remained high in the SF. The variation was significantly influenced by temperature, with lower emission and higher dissolved CH(4) in water occurring at lower temperatures. Both the emission and concentration of dissolved CH(4) were also influenced significantly by water quality, wetland design, level of stalk butt left above the water level, etc. The methane flux was explained on the basis of rizhospheric methanogenic and methanotrophic microbial populations. FISH analysis indicated the presence of Type A and Type B methanotrophs in both wetlands, and the methane flux was directly influenced by the quantitative variation in methanogenic and methanotrophic bacteria in both wetlands.

Bacteria↗

PubMed2006

Methane flux and oxidation at two types of intermediate landfill covers.

Methane emissions were measured on two areas at a Florida (USA) landfill using the static chamber technique. Because existing literature contains few measurements of methane emissions and oxidation in intermediate cover areas, this study focused on field measurement of emissions at 15-cm-thick non-vegetated intermediate cover overlying 1-year-old waste and a 45-cm-thick vegetated intermediate cover overlying 7-year-old waste. The 45 cm thick cover can also simulate non-engineered covers associated with older closed landfills. Oxidation of the emitted methane was evaluated using stable isotope techniques. The arithmetic means of the measured fluxes were 54 and 22 g CH(4) m(-2)d(-1) from the thin cover and the thick cover, respectively. The peak flux was 596 g m(-2)d(-1) for the thin cover and 330 g m(-2)d(-1) for the thick cover. The mean percent oxidation was significantly greater (25%) at the thick cover relative to the thin cover (14%). This difference only partly accounted for the difference in emissions from the two sites. Inverse distance weighing was used to describe the spatial variation of flux emissions from each cover type. The geospatial mean flux was 21.6 g m(-2)d(-1) for the thick intermediate cover and 50.0 g m(-2)d(-1) for the thin intermediate cover. High emission zones in the thick cover were fewer and more isolated, while high emission zones in the thin cover were continuous and covered a larger area. These differences in the emission patterns suggest that different CH(4) mitigation techniques should be applied to the two areas. For the thick intermediate cover, we suggest that effective mitigation of methane emissions could be achieved by placement of individualized compost cells over high emission zones. Emissions from the thin intermediate cover, on the other hand, can be mitigated by placing a compost layer over the entire area.

Air Pollutants↗

PubMed2004

Evidence of anoxic methane oxidation coupled to denitrification.

Denitrification using methane as sole electron donor under anoxic condition was investigated. Sludge produced by a denitrifying reactor using acetate as electron donor was put in contact with methane at partial pressures from 1.8 to 35.7kPa. Nitrate depletion and gaseous nitrogen production were measured. The denitrification rate was independent of the methane partial pressure when superior or equal to 8.8kPa. The nitrate depletion was asymptotic. A denitrification rate of 0.25g NO(3)(-)-Ng(-1) VSSd(-1) was observed at the onset of culturing, followed by a slower and lineal denitrification rate of 4.9x10(-3)g NO(3)(-)-Ng(-1) VSSd(-1). Abiotic nitrate removal or the availability of another carbon source were discarded from control experiments made in the absence of methane or using sterilized inoculum.

Atmospheric Pressure↗

PubMed2005

In situ bioremediation of a cis-dichloroethylene-contaminated aquifer utilizing methane-rich groundwater from an uncontaminated aquifer.

At a trichloroethylene (TCE)-contaminated site in Chikura, Chiba, Japan, TCE had spread over to the first and second aquifers over years. After 8 years of pumping and treatment, finally derivative of TCE, cis-dichloroethylene (c-DCE) remained only in the second aquifer. In this study, feasibility of a low cost in situ bioremediation utilizing groundwater of the third aquifer, which contained natural dissolved methane possibly derived from natural gas field nearby, to stimulate methane-oxidizing bacteria was examined. In vitro experiment showed that a mixture of the groundwater from the second and third aquifers stimulated a growth of methane oxidizing bacteria and enhanced c-DCE degradation. The groundwater of the third aquifer was introduced into the second aquifer in situ. The population of methanotrophs with high V(max) and K(m) for methane uptake increased, resulting in successful degradation of c-DCE at a monitoring well 2m downgradient of the injection well.

Bacteria↗

PubMed2006

Hydrogen and methane production from household solid waste in the two-stage fermentation process.

A two-stage process combined hydrogen and methane production from household solid waste was demonstrated working successfully. The yield of 43 mL H(2)/g volatile solid (VS) added was generated in the first hydrogen production stage and the methane production in the second stage was 500 mL CH(4)/g VS added. This figure was 21% higher than the methane yield from the one-stage process, which was run as control. Sparging of the hydrogen reactor with methane gas resulted in doubling of the hydrogen production. pH was observed as a key factor affecting fermentation pathway in hydrogen production stage. The optimum pH range for hydrogen production in this system was in the range from 5 to 5.5. The short hydraulic retention time (2 days) applied in the first stage was enough to separate acidogenesis from methanogenesis. No additional control for preventing methanogenesis in the first stage was necessary. Furthermore, this study also provided direct evidence in the dynamic fermentation process that, hydrogen production increase was reflected by acetate to butyrate ratio increase in liquid phase.

Acetates↗

PubMed1978

Coenzyme M and methylcobalamin in methane biosynthesis: results of model studies.

Coenzyme M (2-mercaptoethanesulfonate, HS-CH2CH2SO3-) reacts with methylcobalamin nonenzymatically in the pH-range between 6 and 14 to yield the S-methyl derivative (CH3-S-CH2CH2SO3-). In addition, and also at lower pH, methane is produced by reductive cleavage of the Co-C bond. With methylcobaloximes as the methyl group donors, methane production predominates, with insignificant S-methylation. The initial rates of methane production from methylcobaloximes with coenzyme M as the reductant correlate with the rates of methane production from these substrates with active cell extracts of Methanobacterium M.o.H.

Bacteria↗

PubMedDate not supplied

Colonic concentrations of hydrogen and methane following colonoscopic preparation with an oral lavage solution.

Explosions are a rare complication of electrocautery in the colon. The cause of these explosions is ignition of gas containing hydrogen (greater than 4%) or methane (greater than 5%), and oxygen. Hydrogen and methane are products of colonic bacterial metabolism. Oral solutions used for colonoscopy preparation are designed to cleanse the colonic lumen but provide no substrate for fermentation reactions. In 52 patients undergoing colonoscopy, gas samples were collected from the right, transverse, and left colon and analyzed for hydrogen, methane, oxygen, and carbon dioxide. The highest hydrogen and methane concentrations were 0.6% and 0.7%, respectively, less than 16% of the explosive level. Oxygen and carbon dioxide concentrations were close to those of air, suggesting that the bulk of colonic gas was air infused during the procedure. We conclude that the concentrations of explosive gases are well below the hazardous levels when oral colonic cleansing solutions and standard colonoscopic procedures are employed.

Administration, Oral↗

PubMed2002

Methane emissions from a coastal lagoon: Vembanad Lake, West Coast, India.

An attempt has been made to estimate methane fluxes from a tropical coastal wetland the Vembanad Lake, a lagoon along the West Coast of India. It has been found that Vembanad Lake contributes significant amount of methane to the atmosphere. Average emissions varied spatially within the lake. Methane emissions were 193.2 +/- 24.5 mg m(-2) h(-1) at Kumaragam (fresh water) as compared to 9.3 +/- 9.6 mgm(-2) h(-1) at Pullot (brackish water) site. Seasonal variation was significant between pre- and post-monsoons. Soil temperature, time of the day, salinity sediment organic carbon, all control the rate of methane emissions from the Vembanad Lake.

Ecosystem↗

PubMed2005

Community-level analysis: key genes of aerobic methane oxidation.

Aerobic methane-oxidizing bacteria (methanotrophs) are a diverse group of bacteria that are currently represented by 13 recognized genera. They play a major role in the global methane cycle and are widespread in nature with representatives found in soils, freshwater, seawater, freshwater and marine sediments, peat bogs and at extremes of temperature, salinity, and pH. There has been an interest in methanotrophs for their potential in bioremediation processes. Methanotroph diversity and ecology are often studied using the "functional" genes pmoA, mmoX, and mxaF, encoding subunits of the particulate methane monooxygenase, soluble methane monooxygenase, and the methanol dehydrogenase, respectively. This chapter describes methods used to detect and analyze these functional genes.

Alcohol Oxidoreductases↗

PubMed1998

Contributions of solvent-solvent hydrogen bonding and van der Waals interactions to the attraction between methane molecules in water.

The contribution of solvent-solvent hydrogen bonding and van der Waals interactions to the attraction between methane molecules in water was investigated by comparing the potential of mean force (PMF) between two methane molecules in TIP4P water to those in a series of related liquids in which the solvent-solvent interactions were progressively turned off while keeping the solvent-solute interactions unchanged. The magnitude of the attraction between methanes was not significantly changed when the hydrogen bonding interaction between solvent molecules was eliminated and the solvent was maintained in the liquid state by increasing either the pressure or the magnitude of the solvent-solvent van der Waals interaction. However, when solvent-solvent excluded volume interactions were eliminated, the methane molecules interacted no more strongly than in the gas phase. The results are consistent with the idea that the primary contribution of hydrogen bonding to the hydrophobic interaction is to keep water molecules in a liquid state; at constant density, packing interactions rather than hydrogen bonding appear to be critical as suggested by scaled particle theories of solvation. The overall shape of the PMF was, however, changed in the absence of hydrogen bonding, pointing to an influence of hydrogen bonding on the detailed form of the interactions between nonpolar solutes in water. The effects of correlations between the configurations sampled during the Monte Carlo procedure used in the free energy calculations on the estimation of errors was also characterized.

Hydrogen Bonding↗

PubMed2003

Microbial oxidation of methane from old landfills in biofilters.

Landfill gas emissions are among the largest sources of the greenhouse gas methane. For this reason, the possibilities of microbial methane degradation in biofilters were investigated. Different filter materials were tested in two experimental plants, a bench-scale plant (total filter volume 51 l) and a pilot plant (total filter volume 4 m3). Three months after the beginning of the experiment, very high degradation rates of up to 63 g CH4/(m3h) were observed in the bench-scale plant at mean methane concentrations of 2.5% v/v and with fine-grained compost as biofilter material. However, the degradation rates of the compost biofilter decreased in the fifth month of the experiment, probably due to the accumulation of exopolymeric substances formed by the microorganisms. A mixture of compost, peat, and wood fibers showed stable and satisfactory degradation rates around 20 g/(m3h) at mean concentrations of 3% v/v over a period of one year. In this material, the wood fibers served as a structural material and prevented clogging of the biofilter. Extrapolation of the experimental data indicates that biofilters for methane oxidation have to be at least 100 times the volume of biofilters for odor control to obtain the same cleaning efficiency per unit volume flow of feed gas.

Air Pollution↗

PubMed2001

Methane monooxygenase component B mutants alter the kinetics of steps throughout the catalytic cycle.

Component interactions play important roles in the regulation of catalysis by methane monooxygenase (MMO). The binding of component B (MMOB) to the hydroxylase component (MMOH) has been shown in previous studies to cause structural changes in MMOH that result in altered thermodynamic and kinetic properties during the reduction and oxygen binding steps of the catalytic cycle. Here, specific amino acid residues of MMOB that play important roles in the interconversion of several intermediates of the MMO cycle have been identified. Both of the histidine residues in Methylosinus trichosporium OB3b MMOB (H5 and H33) were chemically modified by diethylpyrocarbonate (DEPC). Although the DEPC--MMOB species exhibited only minor changes relative to unmodified MMOB in steady-state MMO turnover, large decreases in the formation rate constants of the reaction cycle intermediates, compound P and compound Q, were observed. The site specific mutants H5A, H33A, and H5A/H33A were made and characterized. H5A and wild type MMOB elicited similar steady-state and transient kinetics, although the mutant caused a slightly lower rate constant for Q formation. Conversely, H33A exhibited a >50-fold decrease in the P formation rate constant, which resulted in slower formation of Q. The kinetics of the double mutant (H5A/H33A) were similar to those of H33A, suggesting that the highly conserved residue, H33, has the most significant effect on the efficient progress of the cycle. Ongoing NMR investigations of residues perturbed by formation of the MMOH-MMOB complex suggested construction of the MMOB N107G/S109A/S110A/T111A quadruple mutant. This mutant was found to elicit a nearly 2-fold increase in specific activity for steady-state MMO turnover of large substrates such as furan and nitrobenzene but caused no similar increase for the physiological substrate, methane. While the quadruple mutant did not have a significant effect on P and Q formation, it caused an almost 3-fold increase in the decay rate constant of Q for furan oxidation and a 2-fold faster product release rate constant for p-nitrophenol resulting from nitrobenzene oxidation. Conversely, this mutant caused the Q decay rate constant to decrease 7-fold for methane oxidation but left the product release step unaffected. These results show for the first time that MMOB exerts influence at late as well as early steps in the catalytic cycle. They also suggest that MMOB plays a critical role in determining the ability of MMO to distinguish between methane and larger substrates.

Amino Acid Sequence↗

PubMed2006

Three-dimensional structure determination of a protein supercomplex that oxidizes methane to formaldehyde in Methylococcus capsulatus (Bath).

The oxidation of methane to methanol in methanotrophs is catalyzed by the enzyme methane monooxygenase (MMO). Two distinct forms of this enzyme exist, a soluble cytoplasmic MMO (sMMO) and a membrane-bound particulate form (pMMO). The active protein complex termed pMMO-C was purified recently from Methylococcus capsulatus (Bath). The complex consists of pMMO hydroxylase and an additional component pMMO-R, which was proposed to be the reductase for the pMMO complex. Further study of this complex has led here to the proposal that the pMMO-R is in fact methanol dehydrogenase, the subsequent enzyme in the methane oxidation pathway by methanotrophs. We describe here the biochemical and biophysical characterization of a stable purified complex of pMMO hydroxylase (pMMO-H) with methanol dehydrogenase (MDH) and report the first three-dimensional (3D) structure, determined by cryoelectron microscopy and single particle analysis to approximately 16 A resolution. The 3D structure reported here provides the first insights into the supramolecular organization of pMMO with MDH. These studies of pMMO-MDH complexes have provided further understanding of the structural basis for the particular functions of the enzymes in this system which might also be of relevance to the complete process of methane oxidation by methanotrophs under high copper concentration in the environment.

Bacterial Proteins↗

PubMed2004

Enhancing activated carbon adsorption of 2-methylisoborneol: methane and steam treatments.

This research investigated methods for tailoring a commercial, lignite-based granular activated carbon (GAC) to enhance its adsorption of 2-methylisoborneol (MIB) from natural water. Tailoring efforts focused on heat treatments in gas environments comprising steam and/or methane, since these gases can alter GAC pore structure and surface chemistry. Heat treatments that combined methane and steam enhanced MIB adsorption considerably, causing a 4-fold improvement (over untreated GAC) in fixed-bed adsorption performance relative to initial MIB breakthrough. These favorable effects, observed in rapid small-scale column tests, occurred following simultaneous and separate (sequential) applications of methane and steam. Moderately low temperature steam treatments also improved MIB uptake in fixed-bed adsorption tests but to a lesser extent (approximately 1.5-fold). In contrast, methane treatments alone, at various temperatures, led to significant carbon deposition within the GAC pore structure. As a result, total pore volume was reduced and MIB adsorption performance declined.

Adsorption↗

PubMed2005

Characterization study and five-cycle tests in a fixed-bed reactor of titania-supported nickel oxide as oxygen carriers for the chemical-looping combustion of methane.

Recent investigations have shown that in the combustion of carbonaceous compounds CO2 and NOx emissions to the atmosphere can be substantially reduced by using a two stage chemical-looping process. In this process, the reduction stage is undertaken in a first reactor in which the framework oxygen of a reducible inorganic oxide is used, instead of the usual atmospheric oxygen, for the combustion of a carbonaceous compound, for instance, methane. The outlet gas from this reactor is mostly composed of CO2 and steam as reaction products and further separation of these two components can be carried out easily by simple condensation of steam. Then, the oxygen carrier found in a reduced state is transported to a second reactor in which carrier regeneration with air takes place at relatively low temperatures, consequently preventing the formation of thermal NOx. Afterward, the regenerated carrier is carried to the first reactor to reinitiate a new cycle and so on for a number of repetitive cycles, while the carrier is able to withstand the severe chemical and thermal stresses involved in every cycle. In this paper, the performance of titania-supported nickel oxides has been investigated in a fixed-bed reactor as oxygen carriers for chemical-looping combustion of methane. Samples with different nickel oxide contents were prepared by successive incipient wet impregnations, and their performance as oxygen carriers was investigated at 900 degrees C and atmospheric pressure in five-cycle fixed-bed reactor tests using pure methane and pure air for the respective reduction and regeneration stages. The evolution of the outlet gas composition in each stage was followed by gas chromatography, and the involved chemical, structural, and textural changes of the carrier in the reactor bed were studied by using different characterization techniques. From the study, it is deduced that the reactivity of these nickel-based oxygen carriers is in the two involved stages and almost independent of the nickel loading. However, in the reduction stage, carbon deposition, from the thermal decomposition of methane, and CO emissions, mainly derived from the partial reduction of titania as support acting as an additional oxygen source, may impose some constraints to the efficiency of the overall chemical-looping combustion process in CO2 capture.

Air Pollutants↗