甲烷厌氧氧化机理及其应用研究进展

甲烷是一种重要的温室气体, 研究证明甲烷厌氧氧化(AOM)对于降低全球甲烷的排放有着重要意义。参与AOM 反应的最终电子受体可分为三类, 即SO2– 4、NO₂^– /NO₃^–以及以Fe³⁺、Cr⁵⁺等为代表的金属离子。本文基于甲烷厌氧氧化过程所利用的电子受体的差别, 结合不同类型AOM 反应微生物的基因型分析, 阐述了AOM 过程的反应机理、相关的微生物种类及其代谢途径。其中对AAA(AOM-associated archaea, 属于ANME-2d)的分离培养, 以及其利用硝酸盐、Fe³⁺、Cr⁵⁺等离子氧化甲烷的研究对认识AOM 反应机理和AOM 的实际应用有很大推动作用。本文还介绍了AOM 过程在环境污染控制领域实际应用中的最新研究进展, 对AOM 的实际应用及其在节能减排上的价值进行展望。AOM 过程的进一步研究对拓宽该过程的工程应用以及对正确认识全球碳、氮、硫循环均有着重要意义。     

亚硝酸盐型甲烷厌氧氧化及其功能微生物研究进展

亚硝酸盐型甲烷厌氧氧化（nitrite-dependent anaerobic methane oxidation,N-DAMO）是耦合氮循环和碳循环的关键环节,主要是由亚硝酸盐型甲烷厌氧氧化菌（Candidatus Methylomirabilis oxyfera）介导完成,对于研究全球氮和碳元素的生物地球化学循环具有重要意义。本文首先总结了国内外N-DAMO的影响因素和在不同自然生态系统中的分布;然后阐述了N-DAMO菌的生理生化特性及其富集培养优化实验和检测技术,最后探讨了N-DAMO技术的应用现状。本综述不仅有助于揭示全球碳氮循环的耦合作用机制,也为N-DAMO反应耦合其他厌氧生物处理过程应用到污水的除碳脱氮上提供了理论依据。     

水生生态系统中金属依赖型甲烷厌氧氧化过程的研究进展

甲烷是一种比CO2更活跃的温室气体,微生物驱动的甲烷厌氧氧化(anaerobic oxidation of methane,AOM)过程对于降低全球甲烷的排放有着重要意义.参与AOM反应的最终电子受体主要分为三类,即硫酸盐、亚硝酸盐/硝酸盐以及以Fe(Ⅲ)、Mn(ⅣV)等为代表的金属离子.可溶性金属物质和不溶性金属矿物...     

厌氧甲烷氧化微生物物质代谢与能量代谢研究进展

甲烷既是一种温室气体,也是一种潜在的能源物质,其源与汇的平衡对地球化学循环及工程应用均有重要意义。厌氧甲烷氧化(anaerobic oxidation of methane,AOM)过程是深海、湿地和农田等自然生境中重要的甲烷汇,在缓解温室气体排放方面发挥了巨大作用。AOM微生物的中枢代谢机制及其能量转化途径则是介导厌氧甲烷氧化耦合其他物质还原的关键所在。因此,本文从电子受体多样性的视角,主要分析了硫酸盐型,硝酸盐/亚硝酸盐型,金属还原型厌氧甲烷氧化微生物的生理生化过程及环境分布,并对近些年发现的新型厌氧甲烷氧化进行了梳理;重点总结了厌氧甲烷氧化微生物细胞内电子传递路径以及胞外电子传递方式;根据厌氧甲烷氧化微生物环境分布及反应特征,就其生态学意义及在污染治理与能源回收方面的潜在应用价值进行了展望。本综述以期深化对厌氧甲烷氧化过程的微生物学认知,并为其潜在的工程应用方向提供新的思路。     

微生物厌氧甲烷氧化反硝化研究进展

厌氧甲烷氧化反硝化过程(Denitrifying anaerobic methane oxidation,DAMO)以甲烷为电子供体进行反硝化作用,在实现废水脱氮处理的同时,可有效削减温室气体甲烷的排放,从而减缓全球温室效应。相关机制研究集中在逆向产甲烷途径耦合反硝化和亚硝酸盐依赖型厌氧甲烷氧化(nitrite-dependent anaerobic methane oxidation,n-damo)两个方面。鉴于厌氧甲烷氧化反硝化过程对全球碳氮物质循环的重要意义,本文对近年来厌氧甲烷氧化反硝化过程的研究进展进行了概述,着重阐述了有关厌氧甲烷氧化反硝化微生物富集培养物,特别是含Candidatus Methylomirabilis oxyfera(M.oxyfera)富集培养物的微生物特性、甲烷氧化反硝化的机理以及影响因子。在此基础上,探讨了厌氧甲烷氧化反硝化过程未来的研究方向和工业化应用前景。     

厌氧消化器中的甲烷氧化菌

自产沼气的厌氧消化器中分离到两株甲烷氧化菌。对这类菌在厌氧消化器中的数量变化及其对产甲烷菌生成甲烷活性的影响作了初步探讨。     

厌氧消化器中的甲烷氧化菌

自产沼气的厌氧消化器中分离到两株甲烷氧化菌。对这类菌在厌氧消化器中的数量变化及其对产甲烷菌生成甲烷活性的影响作了初步探讨。     

甲烷氧化菌及其在环境治理中的应用

甲烷的生物氧化包括好氧氧化和厌氧氧化两种,分别由好氧甲烷氧化菌和厌氧甲烷氧化菌完成.由于该过程是减少自然环境中温室气体甲烷排放的重要途径,越来越受到各国学者的重视.本文主要对当前甲烷氧化菌的研究现状进行了综述,对好氧甲烷氧化菌的种类、参与氧化甲烷的关键酶,厌氧甲烷氧化菌的种类、参与的微生物菌种以及氧化机理进行了论述,并对这两类微生物在温室气体减排、污染物治理、废水生物脱氮、硫及金属元素回收等方面的应用现状及前景进行了分析.     

硝态氮还原型厌氧甲烷氧化的研究进展及展望

以硝态氮为电子受体的甲烷厌氧氧化(Nx-damo)是近年被证实的微生物驱动的地球氮、碳循环机制,对于认识重要元素地球化学循环的微生物驱动机制和自然环境中甲烷的源与汇具有重大意义;在废水生物脱氮及其温室气体减排方面也具有潜在工程应用价值。从功能微生物富集及其影响因素、生理特性、生物代谢的可能机理等方面对Nx-damo的最新进展进行了梳理和讨论;评估了其应用于废水处理的潜力和优势;对未来的研究方向进行了展望,以期推动该领域更广泛的研究,并为其提供有价值的参考。     

硝酸盐和硫酸盐厌氧氧化甲烷途径及氧化菌群

甲烷属于温室气体,厌氧氧化甲烷有效地减少了大气环境中甲烷的含量。依据吉布斯自由能变,以SO42、Mn4+、Fe3+、NO3等作为电子受体,厌氧条件下甲烷可以转化为CO2。重点阐述以SO42和NO3为电子受体时甲烷厌氧氧化的机理、反应发生的环境条件以及甲烷厌氧氧化菌的特点。针对目前研究存在的主要问题,提出了今后的发展方向。SO42为电子受体时,甲烷厌氧氧化的可能途径包括:逆甲烷生成途径、乙酰生成途径以及甲基生成途径。甲烷的好氧或厌氧氧化协同反硝化是以NO3为电子受体的甲烷氧化的可能途径。环境中的甲烷、硫酸盐或硝酸盐的浓度,有机质的数量,以及环境条件对甲烷的厌氧氧化有显著影响。     

Thermophilic anaerobic oxidation of methane by marine microbial consortia

The anaerobic oxidation of methane (AOM) with sulfate controls the emission of the greenhouse gas methane from the ocean floor. AOM is performed by microbial consortia of archaea (ANME) associated with partners related to sulfate-reducing bacteria. In vitro enrichments of AOM were so far only successful at temperatures ⩽25 °C; however, energy gain for growth by AOM with sulfate is in principle also possible at higher temperatures. Sequences of 16S rRNA genes and core lipids characteristic for ANME as well as hints of in situ AOM activity were indeed reported for geothermally heated marine environments, yet no direct evidence for thermophilic growth of marine ANME consortia was obtained to date. To study possible thermophilic AOM, we investigated hydrothermally influenced sediment from the Guaymas Basin. In vitro incubations showed activity of sulfate-dependent methane oxidation between 5 and 70 °C with an apparent optimum between 45 and 60 °C. AOM was absent at temperatures ⩾75 °C. Long-term enrichment of AOM was fastest at 50 °C, yielding a 13-fold increase of methane-dependent sulfate reduction within 250 days, equivalent to an apparent doubling time of 68 days. The enrichments were dominated by novel ANME-1 consortia, mostly associated with bacterial partners of the deltaproteobacterial HotSeep-1 cluster, a deeply branching phylogenetic group previously found in a butane-amended 60 °C-enrichment culture of Guaymas sediments. The closest relatives (Desulfurella spp.; Hippea maritima) are moderately thermophilic sulfur reducers. Results indicate that AOM and ANME archaea could be of biogeochemical relevance not only in cold to moderate but also in hot marine habitats.     

Biogeochemistry and microbial ecology of methane oxidation in anoxic environments: a review

Evidence supporting a key role for anaerobic methane oxidation in the global methane cycle is reviewed. Emphasis is on recent microbiological advances. The driving force for research on this process continues to be the fact that microbial communities intercept and consume methane from anoxic environments, methane that would otherwise enter the atmosphere. Anaerobic methane oxidation is biogeochemically important because methane is a potent greenhouse gas in the atmosphere and is abundant in anoxic environments. Geochemical evidence for this process has been observed in numerous marine sediments along the continental margins, in methane seeps and vents, around methane hydrate deposits, and in anoxic waters. The anaerobic oxidation of methane is performed by at least two phylogenetically distinct groups of archaea, the ANME-1 and ANME-2. These archaea are frequently observed as consortia with sulfate-reducing bacteria, and the metabolism of these consortia presumably involves a syntrophic association based on interspecies electron transfer. The archaeal member of a consortium apparently oxidizes methane and shuttles reduced compounds to the sulfate-reducing bacteria. Despite recent advances in understanding anaerobic methane oxidation, uncertainties still remain regarding the nature and necessity of the syntrophic association, the biochemical pathway of methane oxidation, and the interaction of the process with the local chemical and physical environment. This review will consider the microbial ecology and biogeochemistry of anaerobic methane oxidation with a special emphasis on the interactions between the responsible organisms and their environment. This revised version was published online in August 2006 with corrections to the Cover Date.     

Quinones as terminal electron acceptors for anaerobic microbial oxidation of phenolic compounds

The capacity of anaerobic granular sludge for oxidizing phenoland p-cresol under anaerobic conditions was studied. Phenol and p-cresolwere completely converted to methane when bicarbonate was the only terminal electron acceptor available. When the humic model compound, anthraquinone-2,6-disulfonate, was included as an alternative electron acceptor in the cultures, the oxidation of the phenolic compounds was coupled to the reduction of the model humic compound to its corresponding hydroquinone, anthrahydroquinone-2,6-disulfonate. These results demonstrate for the first time that the anaerobic degradation of phenolic compounds can be coupled to the reduction of quinones as terminal electron acceptor.     

Anaerobic oxidation of methane at different temperature regimes in Guaymas Basin hydrothermal sediments

Anaerobic oxidation of methane (AOM) was investigated in hydrothermal sediments of Guaymas Basin based on δ¹³C signatures of CH₄, dissolved inorganic carbon and porewater concentration profiles of CH₄ and sulfate. Cool, warm and hot in-situ temperature regimes (15–20 °C, 30–35 °C and 70–95 °C) were selected from hydrothermal locations in Guaymas Basin to compare AOM geochemistry and 16S ribosomal RNA (rRNA), mcrA and dsrAB genes of the microbial communities. 16S rRNA gene clone libraries from the cool and hot AOM cores yielded similar archaeal types such as Miscellaneous Crenarchaeotal Group, Thermoproteales and anaerobic methane-oxidizing archaea (ANME)-1; some of the ANME-1 archaea formed a separate 16S rRNA lineage that at present seems to be limited to Guaymas Basin. Congruent results were obtained by mcrA gene analysis. The warm AOM core, chemically distinct by lower porewater sulfide concentrations, hosted a different archaeal community dominated by the two deep subsurface archaeal lineages Marine Benthic Group D and Marine Benthic Group B, and by members of the Methanosarcinales including ANME-2 archaea. This distinct composition of the methane-cycling archaeal community in the warm AOM core was confirmed by mcrA gene analysis. Functional genes of sulfate-reducing bacteria and archaea, dsrAB, showed more overlap between all cores, regardless of the core temperature. 16S rRNA gene clone libraries with Euryarchaeota-specific primers detected members of the Archaeoglobus clade in the cool and hot cores. A V6-tag high-throughput sequencing survey generally supported the clone library results while providing high-resolution detail on archaeal and bacterial community structure. These results indicate that AOM and the responsible archaeal communities persist over a wide temperature range.     

Trace methane oxidation studied in several Euryarchaeota under diverse conditions

We used (13)C-labeled methane to document the extent of trace methane oxidation by Archaeoglobus fulgidus, Archaeoglobus lithotrophicus, Archaeoglobus profundus, Methanobacterium thermoautotrophicum, Methanosarcina barkeri and Methanosarcina acetivorans. The results indicate trace methane oxidation during growth varied among different species and among methanogen cultures grown on different substrates. The extent of trace methane oxidation by Mb. thermoautotrophicum (0.05 +/- 0.04%, +/- 2 standard deviations of the methane produced during growth) was less than that by M. barkeri (0.15 +/- 0.04%), grown under similar conditions with H(2) and CO(2). Methanosarcina acetivorans oxidized more methane during growth on trimethylamine (0.36 +/- 0.05%) than during growth on methanol (0.07 +/- 0.03%). This may indicate that, in M. acetivorans, either a methyltransferase related to growth on trimethylamine plays a role in methane oxidation, or that methanol is an intermediate of methane oxidation. Addition of possible electron acceptors (O(2), NO(3) (-), SO(4) (2-), SO(3) (2-)) or H(2) to the headspace did not substantially enhance or diminish methane oxidation in M. acetivorans cultures. Separate growth experiments with FAD and NAD(+) showed that inclusion of these electron carriers also did not enhance methane oxidation. Our results suggest trace methane oxidized during methanogenesis cannot be coupled to the reduction of these electron acceptors in pure cultures, and that the mechanism by which methane is oxidized in methanogens is independent of H(2) concentration. In contrast to the methanogens, species of the sulfate-reducing genus Archaeoglobus did not significantly oxidize methane during growth (oxidizing 0.003 +/- 0.01% of the methane provided to A. fulgidus, 0.002 +/- 0.009% to A. lithotrophicus and 0.003 +/- 0.02% to A. profundus). Lack of observable methane oxidation in the three Archaeoglobus species examined may indicate that methyl-coenzyme M reductase, which is not present in this genus, is required for the anaerobic oxidation of methane, consistent with the "reverse methanogenesis" hypothesis.     

Enhancing mainstream nitrogen removal by employing nitrate/nitrite-dependent anaerobic methane oxidation processes

Due to serious eutrophication in water bodies, nitrogen removal has become a critical stage for wastewater treatment plants (WWTPs) over past decades. Conventional biological nitrogen removal processes are based on nitrification and denitrification (N/DN), and are suffering from several major drawbacks, including substantial aeration consumption, high fugitive greenhouse gas emissions, a requirement for external carbon sources, excessive sludge production and low energy recovery efficiency, and thus unable to satisfy the escalating public needs. Recently, the discovery of anaerobic ammonium oxidation (anammox) bacteria has promoted an update of conventional N/DN-based processes to autotrophic nitrogen removal. However, the application of anammox to treat domestic wastewater has been hindered mainly by unsatisfactory effluent quality with nitrogen removal efficiency below 80%. The discovery of nitrate/nitrite-dependent anaerobic methane oxidation (n-DAMO) during the last decade has provided new opportunities to remove this barrier and to achieve a robust system with high-level nitrogen removal from municipal wastewater, by utilizing methane as an alternative carbon source. In the present review, opportunities and challenges for nitrate/nitrite-dependent anaerobic methane oxidation are discussed. Particularly, the prospective technologies driven by the cooperation of anammox and n-DAMO microorganisms are put forward based on previous experimental and modeling studies. Finally, a novel WWTP system acting as an energy exporter is delineated.     

Release of fossil methane from mineral soil particles, and its implication for estimation of methane oxidation in a mineral subsoil

In a preliminary experiment we found that methane evolved from a sandy subsoil during aerobic incubation of shaken soil slurries. In the study presented here the methane was found to be released from the sand particles by mechanical weathering, caused by the grinding effect of the shaking. Large amounts of gas (about 0.5 ml gas g^–1 soil) were extracted by intense grinding of the soil in gas tight serum vials. Methane was the main hydrocarbon in the emitted gas, but also a considerable amount of ethane was present, as well as minor amounts of heavier hydrocarbons (up to C₆). The ¹³C-values of the emitted methane and ethane were –33 and –29 , respectively. Together these results demonstrate a thermogenic origin of the gas. This paper also reports the results of an incubation experiment where possible methane oxidation was looked for. If a possible release of methane is not accounted for, methane oxidation may be overlooked, as illustrated in this paper. Methane consumption was detected only in soil from 40 cm, in contrast to soil sampled at 100 cm and deeper where a slight production was measured. When methane oxidation was inhibited by dimethyl-ether, a significant release of methane was seen. The release was probably caused by chemical weathering. When this methane release was taken into account, methane oxidation was found to be present at all measured depths (40 to 200 cm). Fertilization with urea inhibited the methane oxidation at 40 cm but not at deeper layers. It is hypothesized that ammonia oxidizing bacteria were the main methane oxidizers in this mineral subsoil (deeper than 1 m), and that oxidation of methane might be a survival mechanism for ammonia oxidizers in ammonia limited environments.     

Missing methane emissions from leaves of terrestrial plants

The controversial claim that attached leaves of terrestrial plants emit CH₄ aerobically remains to be corroborated. Here, we report CH₄ fluxes and CO₂ exchange rates for leaves of the C₄ species Zea mays using a high-accuracy traceable online analytical system. In contrast to earlier results for Z. mays , our measurements provide no evidence for substantial aerobic CH₄ emissions from photosynthesizing leaves illuminated with photosynthetically active radiation ( λ =400–700 nm), or from dark-respiring leaves. Preliminary measurements with the same system indicated a similar lack of aerobic CH₄ emissions in the light or dark from leaves of the C₃ species Nicotiana tabacum . These findings are supported by independent high-precision ¹³C-labeling studies that also failed to confirm substantial aerobic CH₄ emissions from plants. Nevertheless, we are not able to exclude the possibility that CH₄ emissions from plants may be linked to nonenzymatic processes with an action spectrum lying outside the wavelength range for photosynthesis.     

Anaerobic methane oxidation potential and bacteria in freshwater lakes: Seasonal changes and the influence of trophic status

Nitrite-dependent anaerobic methane oxidation (n-damo), mainly carried out by n-damo bacteria, is an important pathway for mitigating methane emission from freshwater lakes. Although n-damo bacteria have been detected in a variety of freshwater lakes, their potential and distribution, and associated environmental factors, remain unclear. Therefore, the current study investigated the potential and distribution of anaerobic methanotrophs in sediments from Erhai Lake and Dianchi Lake, two adjacent freshwater lakes in the Yunnan Plateau with different trophic status. Both lakes showed active anaerobic methane oxidation potential and harbored a high density of n-damo bacteria. Based on the n-damo pmoA gene, sediment n-damo bacterial communities mainly consisted of Candidatus Methylomirabils oxyfera and Candidatus Methylomirabils sinica, as well as novel n-damo organisms. Sediment anaerobic methane oxidation potential and the n-damo bacterial community showed notable differences among seasons and between lakes. The environmental variables associated with lake trophic status (e.g. total nitrogen, ammonia nitrogen, nitrate nitrogen, and total organic carbon) might have significant impacts on the anaerobic methane oxidation potential, as well as the abundance and community structure of n-damo bacteria. Therefore, trophic status could determine the n-damo process in freshwater lake sediment.     

Dimethylsulfide and methane thiol in sediment porewater of a Danish estuary

Seasonal variation of dimethylsulfide (DMS) and methane thiol (MSH) concentrations in sediment porewater was determined in a Danish estuary. Dimethylsulfide (DMDS) was never found. Detectable DMS levels of up to 0.1 M were found only in the summer and only within the upper 5 cm of the sediment. The DMS accumulation was probably associated with decomposing fragments of macro-algae in the surface layer. Significant MSH accumulation of up to 1 M was found only in the deep, CH₄-rich sediment below the SO₄ ^2- zone. With depth, a detectable MSH level could thus be observed below the 1 mM SO₄ ^2--isopleth which also marked the SO₄ ^2--CH₄ transition. The transition zone was located deeper in the sediment in winter (20–25 cm depth) than in summer (5–10 cm depth). The absence of MSH in the SO₄ ^2- zone could be due to rapid utilization of the compound by SO₄ ^2--reducing bacteria. A possible involvement of MSH in anaerobic CH₄ oxidation at the transition zone is discussed; CH₄ and sulfide (HS^- form, pH 7) are proposed to form MSH and H₂ which in turn may be metabolized by, e.g. SO₄ ^2--reducing bacteria.