微生物介导的甲烷厌氧氧化过程及其影响因子研究进展

温室气体甲烷减排是全球变化领域的研究热点,甲烷厌氧氧化(anaerobic methane oxidation,AOM)过程是一个以前被忽视的甲烷汇,在调控全球甲烷收支平衡及减缓温室效应等方面扮演着十分重要的角色。AOM微生物以甲烷为唯一电子供体,与硫酸盐(SO42-)、亚硝酸盐(NO2-)/硝酸盐(NO3-)、金属离子(Fe3+、Mn4+、Cr6+)等结合完成氧化还原过程,该过程是耦合碳、氮、硫循环的关键环节。本文系统整理分析了不同AOM类型、发生机理、相关功能微生物类群(ANME-1、ANME-2、ANME-3、NC10、MBG-D)及影响AOM过程的关键调控因子的最新研究进展。结果发现,目前80%以上研究都集中在对最常见电子受体类型(SO42-/NO3-/NO2-/Fe3+/Mn4+)的AOM相关过程,而忽视了潜在的新型电子受体(AQDS/HAs O42-/Cr6+/ClO4-等)的耦合作用过程和相对应的微生物类型及作用机理。对未来AOM研究方向提出展望,以期为研究甲烷厌氧氧化菌在不同生态系统中的生态分布及减缓全球温室气体排放提供新的思路。     

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。     

厌氧氨氧化生物脱氮技术的研究进展

厌氧氨氧化是指在厌氧条件下,厌氧氨氧化混合菌直接以NH4 为电子供体,以NO3-或NO2-为电子受体,将NH4^ 、NO3-或NO2-转变成N2的过程。厌氧氨氧化作为一种新型的污水处理工艺具有较高的理论意义和良好的应用前景。本文主要阐述了厌氧氨氧化生物脱氮技术原理、厌氧氨氧化的可能途径、方法及其应用现状,并且讨论了厌氧氨氧化反应的微生物学机理和厌氧氨氧化工艺的开发,提出了今后研究的主要方向。     

厌氧氨氧化菌特性及其在生物脱氮中的应用

在无分子氧环境中,同时存在NH4^＋和NO2^-时,NH4^＋作为反硝化的无机电子供体,NO2^-作为电子受体,生成氮气,这一过程称为厌氧氨氧化。目前已经发现了3种厌氧氨氧化菌（Brocadia anammoxidans,Kuenenia stuttgartiensis,Scalindua sorokinii）;对厌氧氨氧化菌的细胞色素、营养物质、抑制物、结构特征和生化反应机理的研究表明,厌氧氨氧化菌具有多种代谢能力。基于部分硝化至亚硝酸盐,然后与氨一起厌氧氨氧化,以及厌氧氨氧化菌与好氧氨氧化菌或甲烷菌的协同耦合作用,提出了几种生物脱氮的新工艺（ANAMMOX、SHARON—ANAMMOX、CANON和甲烷化与厌氧氨氧化耦合工艺）。     

电子受体和光照对土壤CH4排放相关微生物功能基因丰度的影响

为探究光照条件下添加不同电子受体对土壤甲烷排放的影响及微生物的响应,本研究在土壤中添加3种电子受体(Fe3+、NO3-、SO42-)共设计8个处理,即黑暗+ Fe3+(DF)、黑暗+NO3-(DN)、黑暗+SO42-(DS)、黑暗+蒸馏水(DCK)、光照+Fe3+(LF)、光照+NO3-(LN)、光照+SO42-(LS...     

内陆湿地与水体甲烷厌氧氧化功能微生物研究进展

内陆湿地与水体(如湖泊、河流、水库等)是温室气体甲烷的重要排放源。微生物介导的甲烷厌氧氧化(anaerobic oxidation of methane,AOM)反应在控制内陆湿地与水体甲烷排放中起着不可忽视的作用,对缓解全球温室效应具有重要意义。内陆湿地与水体易形成缺氧环境,且电子受体的种类和数量繁多,是发生AOM反应的理想生境。近年来,不断有研究表明,内陆湿地与水体中存在多种电子受体(NO₂^-、NO₃^-、SO₄^2-、Fe (III)等)驱动的AOM途径。NC10门细菌和甲烷厌氧氧化古菌(anaerobic methanotrophic archaea,ANME)的一新分支ANME-2d主导了湿地和水体环境中的AOM反应,其中ANME-2d具有根据环境条件选择不同电子受体的潜力。研究系统综述了内陆湿地与水体中不同电子受体驱动的AOM途径及其参与的主要功能微生物类群;分析了AOM反应在控制温室气体甲烷排放中的作用及其环境影响因素;总结了相关功能微生物的分子生物学检测方法及甲烷厌氧氧化活性测定的同位素示踪技术。最后,对未来相关研究方向进行了展望。     

厌氧氨氧化菌的中心代谢研究进展

摘要: 厌氧氨氧化是以NH +4为电子供体,以NO－2为电子受体产生N2的生物反应。厌氧氨氧化菌是厌氧氨氧化过程的执行者,在废水生物脱氮和地球氮素循环中扮演着重要角色。研究厌氧氨氧化菌的代谢特性,将有助于理解厌氧氨氧化过程,开发厌氧氨氧化工艺。厌氧氨氧化菌是化能自养型细菌,以CO2或HCO－3为碳源,并通过偶联NH+4氧化和NO －2还原的生物反应获得能量。在NH+4/NO－2的生物氧化还原反应过程中,检出了中间产物N2H4,但未检出其他中间产物(如NH2OH、NO)。此外,由基因组信息推断,厌氧氨氧化菌     

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。     

反硝化型甲烷厌氧氧化的研究进展

反硝化型甲烷厌氧氧化(denitrifying anaerobic methane oxidation,DAMO)即甲烷厌氧氧化耦合反硝化,是指在厌氧条件下以甲烷作为电子供体,NO2-/NO3-作为电子受体的反硝化过程。甲烷是一种温室气体,其引起的温室效应是等物质量CO2的20~30倍。DAMO过程利用甲烷代替常规碳源进行脱氮,有利于减少温室效应,并改善氮循环。研究发现,Candidatus Methylomirabilis oxyfera细菌和Candidatus Methanoperedens nitroreducens古菌是参与DAMO过程的2类主要功能微生物,前者通过内部好氧机制耦合亚硝酸盐还原与甲烷的厌氧氧化,后者则通过逆向产甲烷途径耦合硝酸盐还原与甲烷的厌氧氧化。本文详细阐述了M.oxyfera细菌和M.nitroreducens古菌细胞内代谢途径,着重总结了甲烷、NO2-/NO3-、反应器构型、温度等因素对DAMO性能的影响。并对DAMO实际应用方面的研究现状做了调研。在DAMO功能微生物作用机制、快速富集及影响因素的进一步深入研究的基础上,推进DAMO污水脱氮工艺的应用是未来的主要研究热点和发展方向。     

Oxidation of methane in the absence of oxygen in lake water samples.

Methane was oxidized to carbon dioxide in the absence of oxygen by water samples from Lake Mendota, Madison, Wis. The anaerobic oxidation of methane did not result in the assimilation of carbon from methane into material precipitable by cold 10% trichloracetic acid. Only samples taken at the suface of the sediment of Lake Mendota were capable of catalyzine the anaerobic oxidation of methane. The rate of methane oxidation in the presence of oxygen was highest in samples taken from near the thermocline. Of the radioactive methane oxidized, 30 to 60% was assimilated into material precipitable by cold 10% trichloroacetic acid during aerobic incubation of the samples. These data support the conclusion that two distinct groups of methane-oxidizing organisms occur in stratifield lakes. Enrichments with acetate and methane as the sole sources of carbon and energy and sulfate as the electron acceptor resulted in the growth of bacteria that oxidize methane. Sulfate, acetate, and methane were all required for growth of enrichments. Acetate was not oxidized to carbon dioxide but was assimilated by cells. Methane was not assimilated but was oxidized to carbon dioxide in the absence of air.     

Oxidation of methane in the absence of oxygen in lake water samples.

Methane was oxidized to carbon dioxide in the absence of oxygen by water samples from Lake Mendota, Madison, Wis. The anaerobic oxidation of methane did not result in the assimilation of carbon from methane into material precipitable by cold 10% trichloracetic acid. Only samples taken at the suface of the sediment of Lake Mendota were capable of catalyzine the anaerobic oxidation of methane. The rate of methane oxidation in the presence of oxygen was highest in samples taken from near the thermocline. Of the radioactive methane oxidized, 30 to 60% was assimilated into material precipitable by cold 10% trichloroacetic acid during aerobic incubation of the samples. These data support the conclusion that two distinct groups of methane-oxidizing organisms occur in stratifield lakes. Enrichments with acetate and methane as the sole sources of carbon and energy and sulfate as the electron acceptor resulted in the growth of bacteria that oxidize methane. Sulfate, acetate, and methane were all required for growth of enrichments. Acetate was not oxidized to carbon dioxide but was assimilated by cells. Methane was not assimilated but was oxidized to carbon dioxide in the absence of air.     

Anaerobic Methane Oxidation: Occurrence and Ecology

Anoxic sediments and digested sewage sludge anaerobically oxidized methane to carbon dioxide while producing methane. This strictly anaerobic process showed a temperature optimum between 25 and 37°C, indicating an active microbial participation in this reaction. Methane oxidation in these anaerobic habitats was inhibited by oxygen. The rate of the oxidation followed the rate of methane production. The observed anoxic methane oxidation in Lake Mendota and digested sewage sludge was more sensitive to 2-bromoethanesulfonic acid than the simultaneous methane formation. Sulfate diminished methane formation as well as methane oxidation. However, in the presence of iron and sulfate the ratio of methane oxidized to methane formed increased markedly. Manganese dioxide and higher partial pressures of methane also stimulated the oxidation. The rate of methane oxidation in untreated samples was approximately 2% of the CH₄ production rate in Lake Mendota sediments and 8% of that in digested sludge. This percentage could be increased up to 90% in sludge in the presence of 10 mM ferrous sulfate and at a partial pressure of methane of 20 atm (2,027 kPa).     

Methane production and sulfate reduction in two Appalachian peatlands

Anaerobic carbon mineralization was evaluated over a 1-year period in two Sphagnum-dominated peatlands, Big Run Bog, West Virginia, and Buckle's Bog, Maryland. In the top 35 cm of peat, mean rates of methane production, anaerobic carbon dioxide production, and sulfate reduction at Big Run Bog were 63,406 and 146 mol L^-1 d^-1, respectively, and at Buckle's Bog were 18, 486 and 104 mol L^-1 d^-1. Annual anaerobic carbon mineralization to methane and carbon dioxide at Big Run Bog and Buckle's Bog was 52.8 and 57.2 mol m^-2, respectively. Rates of methane production were similar to rates reported for other freshwater peatlands, but methane production accounted for only 11.7 and 2.8%, respectively, of the total anaerobic carbon mineralization at these two sites. Carbon dioxide production, resulting substantially from sulfate reduction, dominated anaerobic carbon mineralization. Considerable sulfate reduction despite low instantaneous dissolved sulfate concentrations (typically < 300 mol L^-1 of substrate) was apparently fueled by oxidation and rapid turnover of the reduced inorganic sulfur pool.The coincidence of high sulfate inputs to the Big Run Bog and Buckle's Bog watersheds through acid precipitation with the unexpected importance of sulfate reduction leads us to suggest a new hypothesis: peatlands not receiving high sulfate loading should exhibit low rates of anaerobic decomposition, and a predominance of methane production over sulfate reduction; however, if such peatlands become subjected to high rates of sulfur deposition, sulfate reduction may be enhanced as an anaerobic mineralization pathway with attendant effects on carbon balance and peat accumulation.     

Energetics of Growth of a Defined Mixed Culture of Desulfovibrio vulgaris and Methanosarcina barkeri: Interspecies Hydrogen Transfer in Batch and Continuous Cultures

Interspecies hydrogen transfer was studied in Desulfovibrio vulgaris-Methanosarcina barkeri mixed cultures. Experiments were performed under batch and continuous growth culture conditions. Lactate or pyruvate was used as an energy source. In batch culture and after 30 days of simultaneous incubation, these organisms were found to yield 1.5 mol of methane and 1.5 mol of carbon dioxide per mol of lactate fermented. When M. barkeri served as the hydrogen acceptor, growth yields of D. vulgaris were higher compared with those obtained on pyruvate without any electron acceptor other than protons. In continuous culture, all of the carbon derived from the oxidation of lactate was recovered as methane and carbon dioxide, provided the dilution rate was minimal. Increasing the dilution rate induced a gradual accumulation of acetate, causing acetate metabolism to cease at above μ = 0.05 h⁻¹. Under these conditions all of the methane produced originated from carbon dioxide. The growth yields of D. vulgaris were measured when sulfate or M. barkeri was the electron acceptor. Two key observations resulted from the present study. First, although sulfate was substituted by M. barkeri, metabolism of D. vulgaris was only slightly modified. The coculture-fermented lactate produced equimolar quantities of carbon dioxide and methane. Second, acetogenesis and methane formation from acetate were completely separable.     

Inhibition Experiments on Anaerobic Methane Oxidation

Anaerobic methane oxidation is a general process important in controlling fluxes of methane from anoxic marine sediments. The responsible organism has not been isolated, and little is known about the electron acceptors and substrates involved in the process. Laboratory evidence indicates that sulfate reducers and methanogens are able to oxidize small quantities of methane. Field evidence suggests anaerobic methane oxidation may be linked to sulfate reduction. Experiments with specific inhibitors for sulfate reduction (molybdate), methanogenesis (2-bromoethanesulfonic acid), and acetate utilization (fluoroacetate) were performed on marine sediments from the zone of methane oxidation to determine whether sulfate-reducing bacteria or methanogenic bacteria are responsible for methane oxidation. The inhibition experiment results suggest that methane oxidation in anoxic marine sediments is not directly mediated by sulfate-reducing bacteria or methanogenic bacteria. Our results are consistent with two possibilities: anaerobic methane oxidation may be mediated by an unknown organism or a consortium involving an unknown methane oxidizer and sulfate-reducing bacteria.     

High rates of anaerobic methanotrophy at low sulfate concentrations with implications for past and present methane levels

Despite the importance of the anaerobic oxidation of methane (AOM) to global biogeochemical cycles, the relationship between sulfate concentration and the rate of AOM has not been previously experimentally constrained. Here, we present measurements showing substantial methane oxidation at low sulfate concentrations, with no significant decrease in the rate of AOM until sulfate levels are well below 1 mM. At sulfate levels below 1 mM, there appears to be a strong decoupling of AOM and sulfate reduction, with a (13)C-label transferred from methane to carbon dioxide occurring at a rate almost an order of magnitude faster than the observed rate of sulfate reduction. These results allow for the possibility that high rates of AOM occurred in the Archean oceans and that high rates of AOM may be found in freshwater environments (lakes, rivers, etc.) and deep ocean sediments today.     

Influences of pond geochemistry,temperature, and freeze-thaw on terminal anaerobic processes occurring in sediments of six ponds of the McMurdo Ice Shelf,near Bratina Island,Antarctica

The effects of freeze-thaw, freezing and sediment geochemistry on terminal anaerobic processes occurring in sediments taken from below cyanobacterial mats in meltwater ponds of the McMurdo Ice Shelf in Antarctica were investigated. Depending on the geochemical and physical status of the sediments (i.e., frozen or thawed), as well as passage of sediment through a freeze-thaw cycle, terminal carbon and electron flow shifted in which the proportions of hydrogen and acetate utilized for methanogenesis and sulfate reduction changed. Thus, in low-sulfate (or chloride) sediment which was thawed and incubated at 4 degrees C, total carbon and electron flow were mediated by acetate-driven sulfate reduction and H(2)-driven methanogenesis. When the same sediments were incubated frozen, both methanogenesis and sulfate reduction decreased. However, under these conditions methanogenesis was favored over sulfate reduction, and carbon flow from acetate to methane increased relative to sulfate reduction; >70% of methane was contributed by acetate, and more than 80% of acetate was oxidized by pathways not coupled to sulfate reduction. In high-sulfate pond sediments, sulfate reduction was a major process mediating terminal carbon and electron flow in both unfrozen and frozen incubations. However, as with low-sulfate sediments, acetate oxidation became uncoupled from sulfate reduction with freezing. Geochemical and temperature effects could be expressed by linear models in which the log (methanogenesis to sulfate reduction) was negative log linear with respect to either temperature or the log of the sulfate (or chloride) concentration. From these relationships it was possible to predict the ratio for a given temperature (low-sulfate sediments) or sulfate (chloride) concentration. Small transitory changes, such as elevated sulfate reduction coupled to increased acetate turnover, resulted from application of a freeze-thaw cycle to low-salinity pond sediments. The results demonstrate how ecophysiological processes may change in anaerobic systems under extreme conditions (e.g., freezing) and provide new insights into microbial events occurring under these conditions.