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At least 19 recordsLinked to original sources
PubMed1978

Microbial oxidation of methane and methanol: isolation of methane-utilizing bacteria and characterization of a facultative methane-utilizing isolate.

A methane-utilizing organism capable of growth both on methane and on more complex organic substrates as a sole source of carbon and energy, has been isolated and studied in detail. Suspensions of methane-grown cells of this organism oxidized C-1 compounds (methane, methanol, formaldehyde, formate); hydrocarbons (ethane, propane); primary alcohols (ethanol, propanol); primary aldehydes (acetaldehyde, propionaldehyde); alkenes (ethylene, propylene); dimethylether; and organic acids (acetate, malate, succinate, isocitrate). Suspensions of methanol-or succinate-grown cells did not oxidize methane, ethane, propane, ethylene, propylene, or dimethylether, suggesting that the enzymatic systems required for oxidation of these substrates are induced only during growth on methane. Extracts of methane-grown cells contained a particulate reduced nicotinamide adenine dinucleotide-dependent methane monooxygenase activity. Oxidation of methanol, formaldehyde, and primary alcohols was catalyzed by a phenazine methosulfate-linked, ammonium ion-requiring methanol dehydrogenase. Oxidation of primary aldehydes was catalyzed by a phenazine methosulfate-linked, ammonium ion-independent aldehyde dehydrogenase. Formate was oxidized by a nicotinamide adenine dinucleotide-specific formate dehydrogenase. Extracts of methane-grown, but not succinate-grown, cells contained the key enzymes of the serine pathway, hydroxypyruvate reductase and malate lyase, indicating that the enzymes of C-1 assimilation are induced only during growth on C-1 compounds. Glucose-6-phosphate dehydrogenase was induced during growth on glucose. Extracts of methane-grown cells contained low levels of enzymes of the tricarboxylic acid cycle, including alpha-keto glutarate dehydrogenase, relative to the levels found during growth on succinate.

Alcohol Oxidoreductases↗
PubMed1979

Methane formation and methane oxidation by methanogenic bacteria.

Methanogenic bacteria were found to form and oxidize methane at the same time. As compared to the quantity of methane formed, the amount of methane simultaneously oxidized varied between 0.3 and 0.001%, depending on the strain used. All the nine tested strains of methane producers (Methanobacterium ruminantium, Methanobacterium strain M.o.H., M. formicicum, M. thermoautotrophicum, M. arbophilicum, Methanobacterium strain AZ, Methanosarcina barkeri, Methanospirillum hungatii, and the "acetate organism") reoxidized methane to carbon dioxide. In addition, they assimilated a small part of the methane supplied into cell material. Methanol and acetate also occurred as oxidation products in M. barkeri cultures. Acetate was also formed by the "acetate organism," a methane bacterium unable to use methanogenic substrates other than acetate. Methane was the precursor of the methyl group of the acetate synthesized in the course of methane oxidation. Methane formation and its oxidation were inhibited equally by 2-bromoethanesulfonic acid. Short-term labeling experiments with M. thermoautotrophicum and M. hungatii clearly suggest that the pathway of methane oxidation is not identical with a simple back reaction of the methane formation process.

Acetates↗
PubMed2002

Methane emissions of differently fed dairy cows and corresponding methane and nitrogen emissions from their manure during storage.

This study investigated the effects of supplementing 40 g lauric acid (C12) kg(-1) dry matter (DM) in feed on methane emissions from early-lactating dairy cows and the associated effects on methane, nitrous oxide and ammonia release from the manure during storage. Stearic acid (C18), a fatty acid without assumed methane-suppressing potential in the digestive tract of ruminants, was added at 40 g kg(-1) DM to a control diet. The complete feed consisted of forage and concentrate in a ratio of 1.5:1 (DM basis). The manure was stored for 14 weeks either as complete slurry or, separately, as urine-rich slurry and farmyard manure representing two common storage systems. Methane release of the cows, as measured in respiratory chambers, was lower with C12 by about 20%, but this was mostly resulting from a reduced feed intake and, partly, from a lower rate of fibre digestion. As milk yield declined less than feed intake, methane emission per kg of milk was significantly lower with C12 (11.4 g) than with C18 (14.0 g). Faeces of C12-fed cows had a higher proportion of undigested fibre and accordingly methane release from their manure was higher compared with the manure obtained from the C18-fed cows. Overall, manure-derived methane accounted for 8.2% and 15.4% of total methane after 7 and 14 weeks of storage, respectively. The evolution of methane widely differed between manure types and dietary treatments, with a retarded onset of release in complete slurry particularly in the C12 treatment. Emissions of nitrous oxide were lower in the manures from the C12 treatment. This partially compensated for the higher methane release from the C12 manure with respect to the greenhouse gas potential. The total greenhouse gas potential (cow and manure together) accounted for 8.7 and 10.5 kg equivalents of CO2 cow(-1) d(-1) with C12 and C18, respectively. At unaffected urine-N proportion ammonia and total nitrogen losses from stored manure were lower with C12 than with C18 corresponding to the differences in feed and nitrogen intake. The present results suggest that manure storage significantly contributes to total methane emission from dairy husbandry, and that the identification of effective dietary mitigation strategies has to consider both the digestive tract of the animals and the corresponding manure.

Ammonia↗
PubMed2003

The methane production of poultry slaughtering residues and effects of pre-treatments on the methane production of poultry feather.

The biological methane production rate and yield of different poultry slaughtering residues were studied. Poultry offal, blood, and bonemeal were rich in proteins and lipids and showed high methane yields, 0.7-0.9, 0.5, and 0.6-0.7 m3 kg(-1) volatile solids(added), respectively (270-340, 100, and 150-170 m3 ton(-1) wet weight). Blood and bonemeal produced methane rapidly, whereas the methane production of offal was more delayed probably due to long-chain fatty acid inhibition. The length of delay depended on the source and concentration of inoculum and incubation temperature, sewage sludge at 35 degrees C having the shortest delay of a few days, while granular sludge did not produce methane within 94 days of incubation. Feather showed a somewhat lower methane yield, 0.21 m3 kg(-1) volatile solids(added) (50 m3 ton(-1) wet weight). Combined thermal (120 degrees C, 5 min) and enzymatic (commercial alkaline endopeptidase, 2-10 g l(-1)) pre-treatments increased its methane yield by 37 to 51%. Thermal (70-120 degrees C, 5-60 min), chemical (NaOH 2-10 g l(-1), 2-24 h), and enzymatic pre-treatments were less effective, with methane yield increasing by 5 to 32%. Based on the present results, anaerobic digestion of the studied poultry slaughtering residues appears a promising possibility because of the high methane yield and nitrogen content of these residues (8 to 14% N of total solids), whereas pre-treatments were shown to improve the methane production of feather.

Abattoirs↗
PubMed1975

Oxidation of C1 Compounds by Particulate fractions from Methylococcus capsulatus: distribution and properties of methane-dependent reduced nicotinamide adenine dinucleotide oxidase (methane hydroxylase).

Cell-free particulate fractions of extracts from the obligate methylotroph Methylococcus capsulatus catalyze the reduced nicotinamide adenine dinucleotide (NADH) and O2-dependent oxidation of methane (methane hydroxylase). The only oxidation product detected was formate. These preparations also catalyze the oxidation of methanol and formaldehyde to formate in the presence or absence of phenazine methosulphate with oxygen as the terminal electron acceptor. Methane hydroxylase activity cannot be reproducibly obtained from disintegrated cell suspensions even though the whole cells actively respired when methane was presented as a substrate. Varying the disintegration method or extraction medium had no significant effect on the activities obtained. When active particles were obtained, hydroxylase activity was stable at 0 C for days. Methane hydroxylase assays were made by measuring the methane-dependent oxidation of NADH by O2. In separate experiments, methane consumption and the accumulation of formate were also demonstrated. Formate is not oxidized by these particulate fractions. The effects of particle concentration, temperature, pH, and phosphate concentration on enzymic activity are described. Ethane is utilized in the presence of NADH and O2. The stoichiometric relationships of the reaction(s) with methane as substrate were not established since (i) the presumed initial product, methanol, is also oxidized to formate, and (ii) the contribution that NADH oxidase activity makes to the observed consumption of reactants could not be assessed in the presence of methane. Studies with known inhibitors of electron transport systems indicate that the path of electron flow from NADH to oxygen is different for the NADH oxidase, methane hydroxylase, and methanol oxidase activities.

Bacteria↗
PubMed1966

A methane-dependent coccus, with notes on classification and nomenclature of obligate, methane-utilizing bacteria.

Foster, J. W. (The University of Texas, Austin), and Richard H. Davis. A methane-dependent coccus, with notes on classification and nomenclature of obligate, methane-utilizing bacteria. J. Bacteriol. 91:1924-1931. 1966.-A new coccus-shaped bacterium capable of aerobic growth at the expense of methane or methanol in a mineral salts medium is described. The organism did not grow at the expense of any of the conventional substrates or homologous hydrocarbons tested. It is gram-negative, nonmotile, and thermotolerant. It grows well at 50 C, optimally at 37 C, but does not grow at 55 C. The cells are encapsulated and have a characteristic diplococcoid arrangement. Washed, "resting-cell" suspensions oxidized certain primary alcohols and short-chain alkanes, an example of "nongrowth oxidation." Of the methane-C utilized, 86% was "fixed" in organic form; the remainder was oxidized to CO(2). The guanine-cytosine content of the extracted deoxyribonucleic acid was 62.5%. Obligate methane-utilizing bacteria are considered as "one-carbon" organisms rather than hydrocarbon utilizers. The assimilation pathway in the obligate methane-methanol bacteria is different from that in the facultative methanol utilizers. Nomenclatural problems arising from the use of the prefix "Methano-" to denote both bacteria that oxidize methane and bacteria that produce methane are discussed. The obligate, one-carbon, methane-methanol bacteria are considered as "methyl" utilizers, and the prefix "Methylo-" is suggested as a solution to the problem of generic cognomens. "Methylococcus capsulatus" gen. n., sp. n. is the name proposed for the new methane coccus.

Bacteria↗
PubMed2001

Stable methane hydrate above 2 GPa and the source of Titan's atmospheric methane.

Methane hydrate is thought to have been the dominant methane-containing phase in the nebula from which Saturn, Uranus, Neptune and their major moons formed. It accordingly plays an important role in formation models of Titan, Saturn's largest moon. Current understanding assumes that methane hydrate dissociates into ice and free methane in the pressure range 1-2 GPa (10-20 kbar), consistent with some theoretical and experimental studies. But such pressure-induced dissociation would have led to the early loss of methane from Titan's interior to its atmosphere, where it would rapidly have been destroyed by photochemical processes. This is difficult to reconcile with the observed presence of significant amounts of methane in Titan's present atmosphere. Here we report neutron and synchrotron X-ray diffraction studies that determine the thermodynamic behaviour of methane hydrate at pressures up to 10 GPa. We find structural transitions at about 1 and 2 GPa to new hydrate phases which remain stable to at least 10 GPa. This implies that the methane in the primordial core of Titan remained in stable hydrate phases throughout differentiation, eventually forming a layer of methane clathrate approximately 100 km thick within the ice mantle. This layer is a plausible source for the continuing replenishment of Titan's atmospheric methane.

Journal Article↗
PubMed2003

A non-radical mechanism for methane hydroxylation at the diiron active site of soluble methane monooxygenase.

We propose a non-radical mechanism for the conversion of methane into methanol by soluble methane monooxygenase (sMMO), the active site of which involves a diiron active center. We assume the active site of the MMOH(Q) intermediate, exhibiting direct reactivity with the methane substrate, to be a bis(mu-oxo)diiron(IV) complex in which one of the iron atoms is coordinatively unsaturated (five-coordinate). Is it reasonable for such a diiron complex to be formed in the catalytic reaction of sMMO? The answer to this important question is positive from the viewpoint of energetics in density functional theory (DFT) calculations. Our model thus has a vacant coordination site for substrate methane. If MMOH(Q) involves a coordinatively unsaturated iron atom at the active center, methane is effectively converted into methanol in the broken-symmetry singlet state by a non-radical mechanism; in the first step a methane C-H bond is dissociated via a four-centered transition state (TS1) resulting in an important intermediate involving a hydroxo ligand and a methyl ligand, and in the second step the binding of the methyl ligand and the hydroxo ligand through a three-centered transition state (TS2) results in the formation of a methanol complex. This mechanism is essentially identical to that of the methane-methanol conversion by the bare FeO(+) complex and relevant transition metal-oxo complexes in the gas phase. Neither radical species nor ionic species are involved in this mechanism. We look in detail at kinetic isotope effects (KIEs) for H atom abstraction from methane on the basis of transition state theory with Wigner tunneling corrections.

Binding Sites↗
PubMed2000

Two-step concerted mechanism for methane hydroxylation on the diiron active site of soluble methane monooxygenase.

A new concerted mechanism is proposed for the conversion of methane to methanol on intermediate Q of soluble methane monooxygenase (sMMO), the active site of which is considered to involve an Fe2(mu-O)2 diamond core. A hybrid density functional theory (DFT) method is used for our mechanistic study on the important reactivity of the bare FeO+ complex and a diiron model of intermediate Q. The reaction pathway for the methane hydroxylation on the diiron complex is essentially identical to that for the gas-phase reaction by the bare FeO+ complex. Methane is highly activated on the dinuclear iron model through the formation of a methane complex, in which a coordinatively unsaturated iron plays a central role in the bonding interaction between the diiron model and substrate methane. A H atom abstraction via a four-centered transition state and a recombination of the OH and CH3 groups via a three-centered transition state successively occur on the dinuclear iron-oxo species, leading to the formation of a methanol complex that corresponds to intermediate T. These electronic processes take place in a concerted manner. Our mechanism for methane hydroxylation by sMMO is different from the radical mechanism that has been widely accepted for enzymatic hydrocarbon hydroxylation, especially by cytochrome P450.

Binding Sites↗
PubMed2000

Simultaneous monitoring of atmospheric methane and speciated non-methane hydrocarbon concentrations using Peltier effect sub-ambient pre-concentration and gas chromatography.

Sub-ambient trapping, used to pre-concentrate atmospheric samples for non-methane hydrocarbon (NMHC) analysis by gas chromatography, can also be used to measure ambient methane concentrations. Above a sample volume of 40 ml, a dynamic equilibrium is established between ambient and trapped methane allowing for simultaneous quantitative determinations of methane and NMHC. The temperature stability of the trap is critical for quantitative methane analysis and this can be achieved by Peltier effect cooling. Simultaneous measurements of methane and NMHC reduce the equipment required for field trips and can ease the interpretation and modelling of atmospheric data. The feasibility for deployment of the system in remote locations was demonstrated by running the apparatus virtually unattended for a 5-day period. The correlations between the concentrations of methane, ethane and ethene measured during this period are discussed.

Air Pollutants↗
PubMedDate not supplied

[Method for assaying methane monooxygenase activity by measuring methane uptake in the reaction vessel].

The reaction vessel has been designed to measure methane monooxygenase activity. An elastic membrane has been built into one of the walls of the vessel to take liquid samples, avoiding formation of the gaseous phase in the reaction volume. The methane content in the samples is measured in a gas-liquid chromatograph with a flame ionization detector in two ways: 1. by direct measurement of methane in the liquid sample, and 2. by measurement of methane in the gaseous phase after methane diffusion from the liquid sample into the gaseous space of another vessel. The method is simple, sensitive (with a lower limit of 0.1 nMole CH4), and well reproducible. This method permits measurement of the oxidation kinetics of methane and other gaseous hydrocarbons both by intact cells and cell-free preparations of methane oxidizing bacteria.

Chromatography, Gas↗
PubMed2001

Dioxygen Activation and Methane Hydroxylation by Soluble Methane Monooxygenase: A Tale of Two Irons and Three Proteins A list of abbreviations can be found in Section 7.

Methanotrophic bacteria are capable of using methane as their sole source of carbon and energy. The first step in methane metabolism, the oxidation of methane to methanol, is catalyzed by a fascinating enzyme system called methane monooxygenase (MMO). The selective oxidation of the very stable C-H bond in methane under ambient conditions is a remarkable feat that has not yet been repeated by synthetic catalysts and has attracted considerable scientific and commercial interest. The best studied MMO is a complex enzyme system that consists of three soluble protein components, all of which are required for efficient catalysis. Dioxygen activation and subsequent methane hydroxylation are catalyzed by a hydroxylase enzyme that contains a non-heme diiron site. A reductase protein accepts electrons from NADH and transfers them to the hydroxylase where they are used for the reductive activation of O(2). The third protein component couples electron and dioxygen consumption with methane oxidation. In this review we examine different aspects of catalysis by the MMO proteins, including the mechanisms of dioxygen activation at the diiron site and substrate hydroxylation by the activated oxygen species. We also discuss the role of complex formation between the different protein components in regulating various aspects of catalysis.

Journal Article↗
PubMed1976

Stoichiometry of methane oxidation in the methane-oxidizing strain M 102 under the influence of various CH4/O2 mixtures.

In laboratory-scale experiments with growing cells of the obligate methane-oxidizing strain M 102, an overall molar gas turnover ratio of the order given below could be postulated: 1 CH4+1--1.2 O2=0.3 CO2+water. Expectations that the optimal gas mixture of methane and oxygen should lie within the range of this stoichiometric consumption ratio have been verified in fermenter 5 1 batch culture experiments. The optimal range of methane-oxygen mixture, found under the experimental conditions described, is based on the estimated growth parameters as generation and doubling times, yield coefficients related to methane and oxygen, and the efficiency of methane metabolism as indicated in the absolute amounts of CH4, O2, and CO2 turned over. The mentioned stoichiometric relation of 1 CH4:1--1.202 did not change with varying the composition, i.e. the partial pressures of CH4 and O2 introduced as a mixture to the cells. The efficiency of methane oxidation was obviously influenced and decreased markedly when deviating from the broad optimal range of CH4/O2 mixtures. With non-growing cells, on the other hand, the stoichiometric relation showed a considerable shift (1:1.4--1.8 CH4:O2) with a clear tendency towards more O2 consumption. The oxidation potential of growing cells, seems then to have a linear interdependence to the substrate concentrations, i.e. partial pressures.

Bacteria↗
PubMed1999

Role of multiple gene copies in particulate methane monooxygenase activity in the methane-oxidizing bacterium Methylococcus capsulatus Bath.

Genes for the subunits of particulate methane monooxygenase, PmoABC, have been sequenced from the gamma-proteobacterial methanotroph Methylococcus capsulatus Bath. M. capsulatus Bath contains two complete copies of pmoCAB, as well as a third copy of pmoC. The two pmoCAB regions were almost identical at the nucleotide sequence level, differing in only 13 positions in 3183 bp. At the amino acid level, each translated gene product contained only one differing residue in each copy. However, the pmoC3 sequence was more divergent from the two other pmoC copies at both the far N-terminus and far C-terminus. Chromosomal insertion mutations were generated in all seven genes. Null mutants could not be obtained for pmoC3, suggesting that it may play an essential role in growth on methane. Null mutants were obtained for pmoC1, pmoC2, pmoA1, pmoA2, pmoB1 and pmoB2. All of these mutants grew on methane, demonstrating that both gene copies were functional. Copy 1 mutants showed about two-thirds of the wild-type whole-cell methane oxidation rate, while copy 2 mutants showed only about one-third of the wild-type rate, indicating that both gene copies were necessary for wild-type particulate methane monooxygenase activity. It was not possible to obtain double null mutants that were defective in both pmo copies, which may indicate that some expression of pMMO is important for growth.

Amino Acid Sequence↗
PubMed2001

Methane oxidation in three Alberta soils: influence of soil parameters and methane flux rates.

Current concern over the potentially negative impacts of climate change has brought attention to anthropogenic sources of methane, a primary greenhouse gas. Two such emission sources are methane leakage at heavy oil wells and sanitary landfills. At both of these sources, substantial quantities of methane could potentially be oxidised by methanotrophic microbes living in soils. Optimisation of this phenomenon may serve as an inexpensive technique for reducing methane emissions. Soil column and batch incubation experiments were performed on a landfill loam, an agricultural loam and a sedge peat to gain a better quantitative understanding of the biological and physical processes limiting CH4 oxidation in soils that undergo the freeze-thaw cycles associated with northern climates. Moisture content emerged as a critical variable that can limit a soil's CH4 oxidation potential. For example, the oxidation rate of the agricultural soil was seen to increase by an order of magnitude after increasing its moisture content from 6% to 10% of its dry weight.

Alberta↗
PubMed1981

Ruminant methane delta(13C/12C) values: relation to atmospheric methane.

The delta(13C/12C)--values of methane produced by fistulated steers, dairy cattle, and wethers, and dairy and beef cattle herds show a bimodal distribution that appears to be correlated with the plant type (C3 or C4, that is, producing either a three- or a four-carbon acid in the first step of photosynthesis) consumed by the animals. These results indicate that cattle and sheep, on a global basis, release methane with an average delta(13C/12C) value of -60 and -63 per mil, respectively. Together they are a source of atmospheric methane whose delta(13C/12C) is similar to published values for marsh gas and cannot explain the 20 per mil higher values for atmospheric methane.

Animals↗
PubMed1970

Oxygenation of methane by methane-grown Pseudomonas methanica and Methanomonas methanooxidans.

1. Experimental conditions have been found in which small amounts of methanol (approximately 2.5mm) accumulated when washed cell suspensions of methane-grown Pseudomonas methanica and Methanomonas methanooxidans were incubated with methane+oxygen mixtures in Warburg flasks. 2. The methanol formed could be separated completely from water by fractional distillation through glass helices followed by gas chromatography using 20% polyethylene glycol 400 on a Celite 545 support. 3. By using (18)O-enriched oxygen gas the abundance of (18)O in the methanol formed from oxidation of methane was measured with a Perkin-Elmer 270 combined gas chromatograph/mass spectrometer. The results showed that the oxygen in methanol was derived exclusively from gaseous oxygen in both micro-organisms. 4. Control experiments using [(18)O]water in incubation mixtures confirmed that there was negligible incorporation of the oxygen atom from water into methanol.

Bacteria↗