RESUMO
The utilization of bioelectrochemical systems for methane production has attracted increasing attention, but producing methane in these systems requires additional voltage to overcome large cathode overpotentials. To eliminate the need for electrical grid energy, we constructed a microbial reverse-electrodialysis methanogenesis cell (MRMC) by placing a reverse electrodialysis (RED) stack between an anode with exoelectrogenic microorganisms and a methanogenic biocathode. In the MRMC, renewable salinity gradient energy was converted to electrical energy, thus providing the added potential needed for methane evolution from the cathode. The feasibility of the MRMC was examined using three different cathode materials (stainless steel mesh coated with platinum, SS/Pt; carbon cloth coated with carbon black, CC/CB; or a plain graphite fiber brush, GFB) and a thermolytic solution (ammonium bicarbonate) in the RED stack. A maximum methane yield of 0.60 ± 0.01 mol-CH4/mol-acetate was obtained using the SS/Pt biocathode, with a Coulombic recovery of 75 ± 2% and energy efficiency of 7.0 ± 0.3%. The CC/CB biocathode MRMC had a lower methane yield of 0.55 ± 0.02 mol-CH4/mol-acetate, which was twice that of the GFB biocathode MRMC. COD removals (89-91%) and Coulombic efficiencies (74-81%) were similar for all cathode materials. Linear sweep voltammetry and electrochemical impedance spectroscopy tests demonstrated that cathodic microorganisms enhanced electron transfer from the cathode compared to abiotic controls. These results show that the MRMC has significant potential for production of nearly pure methane using low-grade waste heat and a source of waste organic matter at the anode.
Assuntos
Bicarbonatos/química , Fontes de Energia Bioelétrica/microbiologia , Eletricidade , Metano/análise , Methanobacterium/crescimento & desenvolvimento , Biofilmes/crescimento & desenvolvimento , Carbono , Eletrodos , Estudos de Viabilidade , Platina , Salinidade , Soluções , Aço InoxidávelRESUMO
An autotrophic, hydrogenotrophic methanogen, designated strain 17A1(T), was isolated from the profundal sediment of the meromictic Lake Pavin, France. The cells of the novel strain, which were non-motile, Gram-staining-negative rods that measured 2-15 µm in length and 0.2-0.4 µm in width, grew as filaments. Strain 17A1(T) grew in a mineral medium and its growth was stimulated by the addition of yeast extract, vitamins, acetate or rumen fluid. Penicillin, vancomycin and kanamycin reduced growth but did not completely inhibit it. Growth occurred at 14-41 °C (optimum 30 °C), at pH 5.0-8.5 (optimum pH 6.5) and with 0-0.4 M NaCl (optimum 0.1 M). The novel strain utilized H(2)/CO(2) and methanol/H(2) as substrates but not formate, acetate, methylamine/H(2), isobutanol or 2-propanol. Its genomic DNA G+C content was 37.0 mol%. In phylogenetic analyses based on 16S rRNA gene sequences, strain 17A1(T) appeared to be a member of the genus Methanobacterium, with Methanobacterium beijingense 8-2(T) (96.3% sequence similarity) identified as the most closely related established species. Based on phenotypic and phylogenetic data, strain 17A1(T) represents a novel species of methanogen within the genus Methanobacterium, for which the name Methanobacterium lacus sp. nov. is proposed. The type strain is 17A1(T) (=DSM 24406(T)=JCM 17760(T)).
Assuntos
Sedimentos Geológicos/microbiologia , Methanobacterium/classificação , Methanobacterium/isolamento & purificação , Antibacterianos/farmacologia , Técnicas de Tipagem Bacteriana , Composição de Bases , Análise por Conglomerados , Meios de Cultura/química , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , França , Água Doce , Concentração de Íons de Hidrogênio , Methanobacterium/genética , Methanobacterium/crescimento & desenvolvimento , Dados de Sequência Molecular , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Cloreto de Sódio/metabolismo , TemperaturaRESUMO
Electromethanogenesis is the bioreduction of carbon dioxide (CO2) to methane (CH4) utilizing an electrode as electron donor. Some studies have reported the active participation of Methanobacterium sp. in electron capturing, although no conclusive results are available. In this study, we aimed at determining short-time changes in the expression levels of [NiFe]-hydrogenases (Eha, Ehb and Mvh), heterodisulfide reductase (Hdr), coenzyme F420-reducing [NiFe]-hydrogenase (Frh), and hydrogenase maturation protein (HypD), according to the electron flow in independently connected carbon cloth cathodes poised at- 800 mV vs. standard hydrogen electrode (SHE). Amplicon massive sequencing of cathode biofilm confirmed the presence of an enriched Methanobacterium sp. population (>70% of sequence reads), which remained in an active state (78% of cDNA reads), tagging this archaeon as the main methane producer in the system. Quantitative RT-PCR determinations of ehaB, ehbL, mvhA, hdrA, frhA, and hypD genes resulted in only slight (up to 1.5 fold) changes for four out of six genes analyzed when cells were exposed to open (disconnected) or closed (connected) electric circuit events. The presented results suggested that suspected mechanisms for electron capturing were not regulated at the transcriptional level in Methanobacterium sp. for short time exposures of the cells to connected-disconnected circuits. Additional tests are needed in order to confirm proteins that participate in electron capturing in Methanobacterium sp.
Assuntos
Proteínas Arqueais/metabolismo , Fontes de Energia Bioelétrica , Eletrodos , Hidrogenase/metabolismo , Metano/metabolismo , Methanobacterium/enzimologia , Proteínas Arqueais/genética , Dióxido de Carbono , Hidrogenase/genética , Methanobacterium/genética , Methanobacterium/crescimento & desenvolvimentoRESUMO
Methanogenic archaea occupy a functionally important niche in the gut microbial ecosystem of mammals. Our purpose was to quantitatively characterize the dynamics of methanogenesis by integrating microbiology, thermodynamics and mathematical modelling. For that, in vitro growth experiments were performed with pure cultures of key methanogens from the human and ruminant gut, namely Methanobrevibacter smithii, Methanobrevibacter ruminantium and Methanobacterium formicium. Microcalorimetric experiments were performed to quantify the methanogenesis heat flux. We constructed an energetic-based mathematical model of methanogenesis. Our model captured efficiently the dynamics of methanogenesis with average concordance correlation coefficients of 0.95 for CO2, 0.98 for H2 and 0.97 for CH4. Together, experimental data and model enabled us to quantify metabolism kinetics and energetic patterns that were specific and distinct for each species despite their use of analogous methane-producing pathways. Then, we tested in silico the interactions between these methanogens under an in vivo simulation scenario using a theoretical modelling exercise. In silico simulations suggest that the classical competitive exclusion principle is inapplicable to gut ecosystems and that kinetic information alone cannot explain gut ecological aspects such as microbial coexistence. We suggest that ecological models of gut ecosystems require the integration of microbial kinetics with nonlinear behaviours related to spatial and temporal variations taking place in mammalian guts. Our work provides novel information on the thermodynamics and dynamics of methanogens. This understanding will be useful to construct new gut models with enhanced prediction capabilities and could have practical applications for promoting gut health in mammals and mitigating ruminant methane emissions.
Assuntos
Intestinos/microbiologia , Metano/metabolismo , Methanobacterium/metabolismo , Modelos Teóricos , Animais , Biomassa , DNA Arqueal/isolamento & purificação , DNA Arqueal/metabolismo , Metabolismo Energético , Cinética , Methanobacterium/genética , Methanobacterium/crescimento & desenvolvimento , RNA Ribossômico 16S/metabolismo , Ruminantes/microbiologia , TermodinâmicaRESUMO
Anaerobic oxidation of methane (AOM) is an important process for understanding the global flux of methane and its relation to the global carbon cycle. Although AOM is known to be coupled to reductions of sulfate, nitrite, and nitrate, evidence that AOM is coupled with extracellular electron transfer (EET) to conductive solids is relatively insufficient. Here, we demonstrate EET-dependent AOM in a biofilm anode dominated by Geobacter spp. and Methanobacterium spp. using carbon-fiber electrodes as the terminal electron sink. The steady-state current density was kept at 11.0 ± 1.3 mA/m2 in a microbial electrochemical cell, and isotopic experiments supported AOM-EET to the anode. Fluorescence in situ hybridization images and metagenome results suggest that Methanobacterium spp. may work synergistically with Geobacter spp. to allow AOM, likely by employing intermediate (formate or H2)-dependent inter-species electron transport. Since metal oxides are widely present in sedimentary and terrestrial environments, an AOM-EET niche would have implications for minimizing the net global emissions of methane.
Assuntos
Geobacter/crescimento & desenvolvimento , Metano/química , Methanobacterium/crescimento & desenvolvimento , Anaerobiose , Biofilmes , Ciclo do Carbono , Transporte de Elétrons , Geobacter/genética , Geobacter/metabolismo , Sequenciamento de Nucleotídeos em Larga Escala , Metagenômica/métodos , Methanobacterium/genética , Methanobacterium/metabolismo , Oxirredução , Análise de Sequência de DNARESUMO
Certain methanogens have been shown to grow on a Mars soil simulant following a washing procedure using a carbonate buffer. In experiments where liquid fractions from the soil simulant and water mixtures were used in place of the buffer, two out of three of the species demonstrated significantly greater methane production compared to the buffer.
Assuntos
Euryarchaeota/crescimento & desenvolvimento , Marte , Metano/biossíntese , Euryarchaeota/metabolismo , Concentração de Íons de Hidrogênio , Methanobacteriaceae/crescimento & desenvolvimento , Methanobacteriaceae/metabolismo , Methanobacterium/crescimento & desenvolvimento , Methanobacterium/metabolismo , Methanosarcina barkeri/crescimento & desenvolvimento , Methanosarcina barkeri/metabolismo , Microbiologia do SoloRESUMO
Two obligate anaerobic bacterial strains (5-3-Z(T) and Y4-1) were isolated from river sediment and rice field mud, respectively. They degraded straight-chain fatty acids with 4-8 carbon atoms in syntrophic association with methanogens, however, neither tested branch-chain fatty acids nor could benzoate be degraded. The strains formed spores when cocultured with methanogens on butyrate, or when grew on butyrate plus dimethyl sulfoxide (DMSO) in pure culture. The cells were slightly curved rods with Gram-negative cell wall structure, and contained small amount of poly beta-hydroxyalkanoate. The strains could not degrade butyrate alone, nor could use fumarate, sulfate, thiosulfate, sulfur or nitrate as electron acceptors except DMSO for butyrate degradation. The generation time of strain 5-3-Z(T) was about 12h when growing on crotonate at 37 degrees C. The growth of the new strains occurred in the range of pH 5.5-8.4, and of temperature 20-48 degrees C, and at NaCl concentration of 0-700 mM. The G+C content of the genomic DNA of strain 5-3-Z(T) was 40.6mol%. Phylogenetic analysis based on 16S rRNA gene similarity showed the two strains to be a member of species Syntrophomonas erecta (98.4-98.9% sequence similarity), however they differed from the existing strains in both phenotypic and genetic characteristics. Therefore, a new subspecies of S. erecta, S. erecta subsp. sporosyntropha was proposed. The type strain was 5-3-Z(T) (=CGMCC1.5032(T)=JCM13344(T)).
Assuntos
Ácidos Graxos Voláteis/metabolismo , Bacilos Gram-Positivos Formadores de Endosporo/classificação , Bacilos Gram-Positivos Formadores de Endosporo/fisiologia , Microbiologia da Água , Anaerobiose , Composição de Bases , Benzoatos/metabolismo , Butiratos/metabolismo , China , Crotonatos/metabolismo , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Fumaratos/metabolismo , Genes de RNAr , Sedimentos Geológicos/microbiologia , Bacilos Gram-Positivos Formadores de Endosporo/citologia , Bacilos Gram-Positivos Formadores de Endosporo/isolamento & purificação , Concentração de Íons de Hidrogênio , Methanobacterium/crescimento & desenvolvimento , Microscopia Eletrônica de Transmissão , Dados de Sequência Molecular , Nitratos/metabolismo , Filogenia , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Rios/microbiologia , Análise de Sequência de DNA , Homologia de Sequência do Ácido Nucleico , Esporos Bacterianos , Sulfatos/metabolismo , Temperatura , Tiossulfatos/metabolismoRESUMO
The recent discovery of methane in the martian atmosphere is arguably one of the most important discoveries in the field of astrobiology. One possible source of this methane could be a microorganism analogous to those on Earth in the domain Archaea known as methanogens. Methanogens are described as obligately anaerobic, and methods developed to work with methanogens typically include anaerobic media and buffers, gassing manifolds, and possibly anaerobic chambers. To determine if the time, effort, and supplies required to maintain anaerobic conditions are necessary to maintain viability, we compared anaerobically washed cells with cells that were washed in the presence of atmospheric oxygen. Anaerobic tubes were opened, and cultures were poured into plastic centrifuge tubes, centrifuged, and suspended in fresh buffer, all in the presence of atmospheric oxygen. Washed cells from both aerobic and anaerobic procedures were inoculated into methanogenic growth media under anaerobic conditions and incubated at temperatures conducive to growth for each methanogenic strain tested. Methane production was measured at time intervals using a gas chromatograph. In three strains, significant differences were not seen between aerobically and anaerobically washed cells. In one strain, there was significantly less methane production observed following aerobic washing at some time points; however, substantial methane production occurred following both procedures. Thus, it appears that aerobic manipulations for relatively short periods of time with at least a few species of methanogens may not lead to loss of viability. With the discovery of methane in the martian atmosphere, it is likely that there will be an increase in astrobiology-related methanogen research. The research reported here should simplify the methodology.
Assuntos
Euryarchaeota/metabolismo , Metano/biossíntese , Aerobiose , Anaerobiose , Cromatografia Gasosa , Euryarchaeota/crescimento & desenvolvimento , Methanobacterium/crescimento & desenvolvimento , Methanobacterium/metabolismo , Methanococcaceae/crescimento & desenvolvimento , Methanococcaceae/metabolismo , Methanosarcina barkeri/crescimento & desenvolvimento , Methanosarcina barkeri/metabolismo , Fatores de TempoRESUMO
The relatively recent discoveries that liquid water likely existed on the surface of past Mars and that methane currently exists in the martian atmosphere have fueled the possibility of extant or extinct life on Mars. One possible explanation for the existence of the methane would be the presence of methanogens in the subsurface. Methanogens are microorganisms in the domain Archaea that can metabolize molecular hydrogen as an energy source and carbon dioxide as a carbon source and produce methane. One factor of importance is the arid nature of Mars, at least at the surface. If one is to assume that life exists below the surface, then based on the only example of life that we know, liquid water must be present. Realistically, however, that liquid water may be seasonal just as it is at some locations on our home planet. Here we report on research designed to determine how long certain species of methanogens can survive desiccation on a Mars soil simulant, JSC Mars-1. Methanogenic cells were grown on JSC Mars-1, transferred to a desiccator within a Coy anaerobic environmental chamber, and maintained there for varying time periods. Following removal from the desiccator and rehydration, gas chromatographic measurements of methane indicated survival for varying time periods. Methanosarcina barkeri survived desiccation for 10 days, while Methanobacterium formicicum and Methanothermobacter wolfeii were able to survive for 25 days.
Assuntos
Euryarchaeota/metabolismo , Meio Ambiente Extraterreno/química , Marte , Dessecação , Euryarchaeota/crescimento & desenvolvimento , Exobiologia , Metano/biossíntese , Methanobacteriaceae/crescimento & desenvolvimento , Methanobacteriaceae/metabolismo , Methanobacterium/crescimento & desenvolvimento , Methanobacterium/metabolismo , Methanosarcina barkeri/crescimento & desenvolvimento , Methanosarcina barkeri/metabolismoRESUMO
Hydrogen (H2) consumption and methane (CH4) production in pure cultures of three different methanogens were investigated during cultivation with 0, 0.2 and 4.21 µM added nickel (Ni). The results showed that the level of dissolved Ni in the anaerobic growth medium did not notably affect CH4 production in the cytochrome-free methanogenic species Methanobacterium bryantii and Methanoculleus bourgensis MAB1, but affected CH4 formation rate in the cytochrome-containing Methanosarcina barkeri grown on H2 and CO2. Methanosarcina barkeri also had the highest amounts of Ni in its cells, indicating that more Ni is needed by cytochrome-containing than by cytochrome-free methanogenic species. The concentration of Ni affected threshold values of H2 partial pressure (pH2) for all three methanogen species studied, with M. bourgensis MAB1 reaching pH2 values as low as 0.1 Pa when Ni was available in amounts used in normal anaerobic growth medium. To our knowledge, this is the lowest pH2 threshold recorded to date in pure methanogen culture, which suggests that M.bourgensis MAB1 have a competitive advantage over other species through its ability to grow at low H2 concentrations. Our study has implications for research on the H2-driven deep subsurface biosphere and biogas reactor performance.
Assuntos
Hidrogênio/farmacologia , Metano/biossíntese , Methanobacterium/metabolismo , Níquel/farmacologia , Biocombustíveis , Reatores Biológicos/microbiologia , Dióxido de Carbono/metabolismo , Meios de Cultura/química , Methanobacterium/crescimento & desenvolvimento , Methanosarcina barkeri/crescimento & desenvolvimento , Methanosarcina barkeri/metabolismo , Técnicas Microbiológicas , Pressão ParcialRESUMO
This investigation evaluated the effectiveness of biochar of different particle sizes in alleviating ammonium (NH4(+)) inhibition (up to 7 g-N/L) during anaerobic digestion of 6 g/L glucose. Compared to the control treatment without biochar addition, treatments that included biochar particles 2-5 mm, 0.5-1 mm and 75-150 µm in size reduced the methanization lag phase by 23.9%, 23.8% and 5.9%, respectively, and increased the maximum methane production rate by 47.1%, 23.5% and 44.1%, respectively. These results confirmed that biochar accelerated the initiation of methanization during anaerobic digestion under double inhibition risk from both ammonium and acids. Furthermore, fine biochar significantly promoted the production of volatile fatty acids (VFAs). Comparative analysis on the archaeal and bacterial diversity at the early and later stages of digestion, and in the suspended, biochar loosely bound, and biochar tightly bound fractions suggested that, in suspended fractions, hydrogenotrophic Methanobacterium was actively resistant to ammonium. However, acetoclastic Methanosaeta can survive at VFAs concentrations up to 60-80 mmol-C/L by improved affinity to conductive biochar, resulting in the accelerated initiation of acetate degradation. Improved methanogenesis was followed by the colonization of the biochar tightly bound fractions by Methanosarcina. The selection of appropriate biochar particles sizes was important in facilitating the initial colonization of microbial cells.
Assuntos
Compostos de Amônio/química , Carvão Vegetal/química , Methanosarcinales/metabolismo , Eliminação de Resíduos Líquidos/métodos , Ácidos/metabolismo , Anaerobiose , Bactérias/crescimento & desenvolvimento , Bactérias/metabolismo , Ácidos Graxos Voláteis/metabolismo , Metano/metabolismo , Methanobacterium/crescimento & desenvolvimento , Methanobacterium/metabolismo , Methanosarcina/crescimento & desenvolvimento , Methanosarcina/metabolismo , Methanosarcinales/crescimento & desenvolvimento , Tamanho da PartículaRESUMO
Two forms (MR1 and MR2) of S-methylcoenzyme M reductase were purified from Methanobacterium thermoautotrophicum (strain delta H) as recently described (Rospert, S., Linder, D., Ellerman, J. and Thauer, R.K. (1990) Eur. J. Biochem. 194, 871-877). MR2 was at least 50-fold more active than MR1, independent of assay conditions. The two forms are spectroscopically similar, but not identical, by UV-visible, magnetic circular dichroism and resonance Raman spectroscopies. MR2 exhibited an EPR signal corresponding to 20% of the enzyme-bound nickel. Strong EPR signals similar to those previously assigned to Ni(I)F430 bound to methylreductase in Methanobacterium thermoautotrophicum (strain Marburg) (Albracht, S.P.J., Ankel-Fuchs, D., Bocher, R., Ellerman, J., Moll, J., Van der Zwann, J.W. and Thauer, R.K. (1988) Biochim. Biophys. Acta 955, 86-102) were observed in MR2-rich, log-phase, as well as in MR1-rich, slow-growing bacteria. Log-phase cells had dramatically different EPR spectra depending on whether they were removed from the fermenter (under gas flow) before or after cooling to 10 degrees C. EPR spectra of slow-growing cells were insensitive to harvesting conditions. The possible biological significance of the alternate form of methylreductase is discussed.
Assuntos
Methanobacterium/enzimologia , Oxirredutases/química , Dicroísmo Circular , Espectroscopia de Ressonância de Spin Eletrônica , Methanobacterium/crescimento & desenvolvimento , Peso Molecular , Oxirredutases/metabolismo , Análise EspectralRESUMO
Schrödinger stated in his landmark book, What is Life?, that life feeds on negative entropy. In this contribution, the validity of this statement is discussed through a careful thermodynamic analysis of microbial growth processes. In principle, both feeding on negative entropy, i.e. yielding products of higher entropy than the substrates, and generating heat can be used by microorganisms to rid themselves of internal entropy production resulting from maintenance and growth processes. Literature data are reviewed in order to compare these two mechanisms. It is shown that entropy-neutral, entropy-driven, and entropy-retarded growth exist. The analysis of some particularly interesting microorganisms shows that enthalpy-retarded microbial growth may also exist, which would signify a net uptake of heat during growth. However, the existence of endothermic life has never been demonstrated in a calorimeter. The internal entropy production in live cells also reflects itself in the Gibbs energy dissipation accompanying growth, which is related quantitatively to the biomass yield. An empirical correlation of the Gibbs energy dissipation in terms of the physico-chemical nature of the growth substrate has been proposed in the literature and can be used to predict the biomass yield approximately. The ratio of enthalpy change and Gibbs energy change can also be predicted since it is shown to be approximately equal to the same ratio of the relevant catabolic process alone.
Assuntos
Bactérias/crescimento & desenvolvimento , Leveduras/crescimento & desenvolvimento , Aerobiose , Anaerobiose , Bactérias/química , Kluyveromyces/crescimento & desenvolvimento , Methanobacterium/crescimento & desenvolvimento , Saccharomyces cerevisiae/crescimento & desenvolvimento , Termodinâmica , Leveduras/químicaRESUMO
Reduction of carbon dioxide to methane by microorganisms attached to electrodes is a promising process in terms of renewable energy storage strategies. However the efficient and specific electrosynthesis of methane by methanogenic archaea on cathodes needs fundamental investigations of the electron transfer mechanisms at the microbe-electrode interface without the addition of artificial electron mediators. Using well-defined electrochemical techniques directly coupled to gas chromatography and surface analysis by scanning electron microscopy, it is shown that a pure culture of the marine lithoautotrophic Methanobacterium-like archaeon strain IM1 is capable to utilize electrons from graphite cathodes for a highly selective production of methane, without hydrogen serving as a cathode-generated electron carrier. Microbial electrosynthesis of methane with cultures of strain IM1 is achieved at a set potential of -0.4V vs. SHE and is characterized by a coulomb efficiency of 80%, with rates reaching 350 nmol d(-1) cm(-2) after 23 days of incubation. Moreover, potential step measurements reveal a biologically catalyzed hydrogen production at potentials more positive than abiotic hydrogen evolution on graphite, indicating that an excessive supply of electrons to strain IM1 results in proton reduction rather than in a further increase of methane production.
Assuntos
Processos Autotróficos , Fontes de Energia Bioelétrica/microbiologia , Metano/biossíntese , Methanobacterium/crescimento & desenvolvimento , Methanobacterium/metabolismo , Minerais/metabolismo , Dióxido de Carbono/metabolismo , Técnicas de Cultura , Eletroquímica , Eletrodos , Metano/químicaRESUMO
The strictly anaerobic, extremely thermophilic methanogens, Methanobacterium thermoautotrophicum Marburg and M. thermoautotrophicum delta H, have been cultivated in liquid culture and on solid medium in screw-top bottles, which permit continuous monitoring of the growth of the microorganisms. We have been able to routinely grow methanogens in medium containing bicarbonate, TRIS or 4-morpholinepropanesulfonic acid (MOPS) buffers and three different sulfur sources (sulfide, sulfite and thiosulfate) at temperatures up to 70 degrees C and at pressures up to 35 psi while monitoring cell density or colony formation.
Assuntos
Técnicas Bacteriológicas , Methanobacterium/crescimento & desenvolvimento , Anaerobiose , Bicarbonatos , Soluções Tampão , Contagem de Células , Meios de Cultura , Morfolinas , Pressão , Temperatura , TrometaminaRESUMO
The membrane potential (delta psi) of whole cells of Methanobacterium thermoautotrophicum strain delta H was estimated under different external conditions using a TPP(+)-sensitive electrode. The results show that the delta psi values of M. thermoautotrophicum at alkaline pHout (8.5) are comparable with delta psi values under slightly acidic conditions (pH 6.8; 230 and 205 mV, respectively). On the other hand, the size of colonies on Petri dishes was remarkably smaller at pH 8.5 than at 6.8. The delta psi was insensitive to relevant ATPase inhibitors. At pH 6.8, the protonophore 3,3',4',5-tetrachlorosalicylanilide (TCS) strongly inhibited delta psi formation and ATP synthesis driven by methanogenic electron transport. On the other hand, at pH 8.5 the CH4 formation and ATP synthesis were insensitive to TCS and a protonophore-resistant delta psi of approximately 150 mV was determined. The finding of a protonophore-resistant delta psi at pH 8.5 indicates that at alkaline pHout these cells can switch from H(+)-energetics to Na(+)-energetics, when the delta [symbol: see text] H+ becomes limited. The results strongly support the hypothesis that at alkaline pHout Na+ ions might fully substitute for H+ in these cells as the coupling ions.
Assuntos
Methanobacterium/química , Adenosina Trifosfatases/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Concentração de Íons de Hidrogênio , Íons , Potenciais da Membrana , Methanobacterium/crescimento & desenvolvimento , Methanobacterium/metabolismo , Oniocompostos , Compostos Organofosforados , Prótons , Salicilanilidas/farmacologia , SódioRESUMO
The microbial communities in 6 granular sludge samples taken from a lab-scale EGSB reactor operated under different organic loading rates (OLR) at 20 degrees C were studied using PCR-DGGE and RTQ-PCR technique. The results indicate that as the OLR increased from 1.5 kg/(m3 x d) to 10.0 kg/(m3 x d), the microbial communities of archaea and eubacteria in the 6 sludge samples are not changed greatly. The archaea population is highly similar among the 6 samples. The Shannon indexes of archaea population are 5.51, 5.88, 5.47, 5.25, 5.32 and 5.11, respectively. Except the seed sludge, the eubacteria population is similar among the other 5 samples. The Shannon indexes of eubacteria population are 2.97, 5.07, 5.44, 6.38, 6.66 and 5.21, respectively. The dominant archaea in the 6 granular samples are Methanobacterium, Methanocorpusculum, Methanosaeta and Methanospirillum. As the OLR of the reactor increased and the operation time elapsed, Methanocorpusculum parvum and Methanospirillum hungatei, both are hydrotrophic methanogens, become the dominant archaea. The archaea content in the samples decreases slightly at the beginning, but increases later and finally reaches to a level obviously higher than that in the seed sludge.
Assuntos
Bactérias Anaeróbias/metabolismo , Reatores Biológicos/microbiologia , Methanobacterium/metabolismo , Methanospirillum/metabolismo , Eliminação de Resíduos Líquidos/métodos , Anaerobiose , Bactérias Anaeróbias/classificação , Bactérias Anaeróbias/crescimento & desenvolvimento , Biodegradação Ambiental , Methanobacterium/crescimento & desenvolvimento , Methanospirillum/crescimento & desenvolvimento , Dinâmica Populacional , Esgotos/microbiologia , TemperaturaRESUMO
Methyl-coenzyme M reductase (MCR) catalyzes the final step of methanogenesis in which coenzyme B and methyl-coenzyme M are converted to methane and the heterodisulfide, CoMS-SCoB. MCR also appears to initiate anaerobic methane oxidation (reverse methanogenesis). At the active site of MCR is coenzyme F430, a nickel tetrapyrrole. This paper describes the reaction of the active MCR(red1) state with the potent inhibitor, 3-bromopropanesulfonate (BPS; I50 = 50 nM) by UV-visible and EPR spectroscopy and by steady-state and rapid kinetics. BPS was shown to be an alternative substrate of MCR in an ionic reaction that is coenzyme B-independent and leads to debromination of BPS and formation of a distinct state ("MCR(PS)") with an EPR signal that was assigned to a Ni(III)-propylsulfonate species (Hinderberger, D., Piskorski, R. P., Goenrich, M., Thauer, R. K., Schweiger, A., Harmer, J., and Jaun, B. (2006) Angew. Chem. Int. Ed. Engl. 45, 3602-3607). A similar EPR signal was generated by reacting MCR(red1) with several halogenated sulfonate and carboxylate substrates. In rapid chemical quench experiments, the propylsulfonate ligand was identified by NMR spectroscopy and high performance liquid chromatography as propanesulfonic acid after protonolysis of the MCR(PS) complex. Propanesulfonate formation was also observed in steady-state reactions in the presence of Ti(III) citrate. Reaction of the alkylnickel intermediate with thiols regenerates the active MCR(red1) state and eliminates the propylsulfonate group, presumably as the thioether. MCR(PS) is catalytically competent in both the generation of propanesulfonate and reformation of MCR(red1). These results provide evidence for the intermediacy of an alkylnickel species in the final step in anaerobic methane oxidation and in the initial step of methanogenesis.
Assuntos
Ácidos Alcanossulfônicos/metabolismo , Oxirredutases/química , Ácidos Alcanossulfônicos/química , Sítios de Ligação , Catálise , Cromatografia Líquida de Alta Pressão , Espectroscopia de Ressonância de Spin Eletrônica , Cinética , Espectroscopia de Ressonância Magnética , Methanobacterium/enzimologia , Methanobacterium/crescimento & desenvolvimento , Níquel/química , Níquel/metabolismo , Oxirredução , Oxirredutases/metabolismo , Especificidade por SubstratoRESUMO
A spore-forming, anaerobic, syntrophic fatty-acid-oxidizing bacterium, strain 19J-3(T), was isolated from a distilled-spirit-fermenting cellar in Hebei Province, China. The cells were slightly curved rods with a spore at the end of the cell. The optimal temperature for growth was around 37 degrees C and growth occurred in the range 25-45 degrees C. The pH range for growth was 6.5-8.5 and the optimum pH was 7.0-7.5. Crotonate was the only substrate that allowed the strain to grow in pure culture. However, the strain could oxidize saturated fatty acids with four to nine carbon atoms syntrophically in co-culture with Methanobacterium formicicum DSM 1535(T). The strain was not able to utilize sulfate, sulfite, thiosulfate, DMSO, nitrate, fumarate or Fe(III) as electron acceptor. The DNA base composition was 48.8 mol% G+C. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain 19J-3(T) was related to members of the family Syntrophomonadaceae and most closely to Syntrophospora bryantii DSM 3014(T) (94.3 % similarity) and Syntrophomonas wolfei subsp. wolfei DSM 2245(T) (93.6 % similarity). Considering the phylogenetic relationship and phenotypic features, strain 19J-3(T) (=CGMCC 1.5041(T)=JCM 13582(T)) is designated as the type strain of a novel species of the genus Syntrophomonas, Syntrophomonas cellicola sp. nov. Based on the close phylogenetic relationship between the genera Syntrophospora and Syntrophomonas, the presence of sporulation-specific genes in the genome of Syntrophomonas wolfei subsp. wolfei DSM 2245(T) and the description of a spore-forming member of Syntrophomonas, 'Syntrophomonas erecta subsp. sporosyntropha', we propose the assignment of Syntrophospora bryantii to the genus Syntrophomonas as Syntrophomonas bryantii comb. nov.