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。     

导电材料强化微生物直接种间电子传递产甲烷的研究进展

厌氧条件下,微生物可以通过厌氧代谢产生甲烷(CH_4),由此衍生的厌氧消化技术可实现能源的回收利用。产CH_4的关键步骤是刺激发酵细菌和产甲烷古菌之间的有效电子转移,电活性微生物可以取代传统的氢/甲酸盐实现直接种间电子传递,其电子传递效率更高。添加导电材料可以促进直接种间电子传递并提高CH_4产率,是一种更有效的强化电子传递方式。本文在梳理直接种间电子传递发展和机理的基础上,综述了常见的促进直接种间电子传递的碳基和铁基导电材料,对其结构特征、电子传递机理、强化产CH_4和中间产物消耗等方面进行了系统总结。旨在为导电材料促进直接种间电子传递的研究提供参考,并探讨了未来可能的研究方向。     

微生物互营产甲烷过程中的种间电子传递

甲烷作为全球第二大温室气体,是典型的可再生清洁能源,也是碳循环中的重要物质组成。大气中约74%的甲烷由产甲烷古菌和其他微生物的互营产生,种间电子传递(interspecies electron transfer, IET)是微生物菌群降低热力学能垒、实现互营产甲烷的核心过程。IET可分为间接种间电子传递(mediated interspecies electron transfer,MIET)和直接种间电子传递(direct interspecies electron transfer, DIET)两种类型,其中MIET依赖氢气、甲酸等载体完成电子的远距离传输,而DIET则依赖导电菌毛、细胞色素c等膜蛋白,通过微生物的直接接触实现电子传递。本文将从IET的研究历程出发,从电子传递机制、微生物种类、生态多样性等方面对微生物互营产甲烷过程中的两种IET类型进行比较,最后对未来待探索的方向进行展望。本综述有助于加深对微生物互营产甲烷过程中IET的理解,为解决由甲烷引发的全球气候变暖等生态问题提供理论支撑。     

金属还原菌希瓦氏菌厌氧呼吸能力及其在环境修复中的研究进展

Shewanella oneidensis MR-1是一种模式金属还原菌,它能够在厌氧条件下,将多种金属化合物和人工合成染料等作为电子受体还原代谢。因此,该菌常常被用于生态修复等研究。厌氧条件下,S.oneidensis MR-1能够将细胞质内或细胞内膜产生的电子通过定位于细胞内膜、细胞膜周质和细胞外膜上的c-血红色素蛋白或还原酶所组成的具有多样性的电子传递系统,最终传递到存在于细菌细胞外环境中的电子受体。通过对多种电子传递过程的介绍,进一步阐明其对污染物修复和纳米材料合成的机理,从而为未来对该类微生物的利用和开发提供更为充分的理论依据。     

产电微生物Shewanella菌厌氧呼吸代谢网络研究进展

以模式菌株Shewanella oneidensis MR-1为代表的Shewanella菌属产电微生物广泛分布于自然水体环境中。作为兼性厌氧菌,Shewanella菌除了能进行有氧呼吸外,还能利用多种电子受体进行厌氧呼吸。通过多种细胞色素所组成的复杂电子传递网络,Shewanella菌不仅能利用渗入到周质空间的可溶性电子受体进行厌氧呼吸,更为特殊的是其能够借助电子的跨膜传递实现对胞外不溶性电子受体的异化还原代谢。本文概述了近年来Shewanella菌厌氧代谢途径的研究进展,探讨电子传递网络对Shewanella菌呼吸多样性及环境适应性的影响。     

陆地生态系统甲烷产生和氧化过程的微生物机理

陆地生态系统存在许多常年性或季节性缺氧环境,如:湿地、水稻土、湖泊沉积物、动物瘤胃、垃圾填埋场和厌氧生物反应器等。每年有大量有机物质进入这些环境,在缺氧条件下发生厌氧分解。甲烷是有机质厌氧分解的最终产物。产生的甲烷气体可通过缺氧-有氧界面释放到大气,产生温室效应,是重要的温室气体。产甲烷过程是缺氧环境中有机质分解的核心环节,而甲烷氧化是缺氧-有氧界面的重要微生物过程。甲烷的产生和氧化过程共同调控大气甲烷浓度,是全球碳循环不可分割的组成部分。对陆地生态系统甲烷产生和氧化过程的微生物机理研究进展进行了概要回顾和综述。主要内容包括:新型产甲烷古菌即第六和第七目产甲烷古菌和嗜冷嗜酸产甲烷古菌的发现;短链脂肪酸中间产物互营氧化过程与直接种间电子传递机制;新型甲烷氧化菌包括厌氧甲烷氧化菌和疣微菌属好氧甲烷氧化菌的发现;甲烷氧化菌生理生态与环境适应的新机制。这些研究进展显著拓展了人们对陆地生态系统甲烷产生和氧化机理的认识和理解。随着新一代土壤微生物研究技术的发展与应用,甲烷产生和氧化微生物研究领域将面临更多机遇和挑战,对未来发展趋势做了展望。     

基于细胞色素c的胞外电子传递过程

电活性微生物具有独特的胞外电子传递功能,在地球化学循环和环境污染修复中起着重要作用。细胞色素c在电活性微生物胞外电子传递过程中扮演了重要角色,不仅参与直接电子传递途径,还参与电子媒介介导的间接电子传递。其电子传递功能不仅对地球环境中铁、锰、碳等元素的循环具有重要作用,还应用于能源生产、废水处理、生物修复等众多领域,具有良好的应用潜力。本文以电活性微生物的2个模式菌属(希瓦氏菌属和地杆菌属)为例,综述了电活性微生物将电子由胞内转移至胞外的方式和途径,详细阐述了细胞色素c在该胞外电子传递过程中的重要作用,总结了细胞色素c介导的胞外电子传递过程所涉及的分析方法,并对微生物胞外电子传递未来的研究方向提出了展望。     

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。     

典型胞外呼吸细菌的胞内电子转移机制研究进展

胞外呼吸在污染物的降解转化和微生物产电过程中具有重要作用。微生物进行胞外呼吸时,其电子受体多以固态形式存在于胞外,氧化产生的电子必须通过电子传递链从胞内经细胞周质转移到外膜。S.oneidensis MR-1与G.Sulfurreducens作为微生物燃料电池中最常用的模式菌株,是现阶段研究最深入和系统的胞外呼吸细菌,其胞内电子传递过程目前研究最为清楚。这两种胞外呼吸细菌的电子传递需多种细胞色素c的参与,S.oneidensis MR-1位于内膜及周质上的细胞色素c-Cym A和MtrA可将电子由内膜上的醌池通过周质到外膜蛋白MtrC和OmcA,MtrC和OmcA接收电子后可直接还原胞外受体,Type Ⅱ secretion system对外膜蛋白中的MtrC和OmcA起到了转运及定位的作用。而在G.sulfurreducens中,电子由MacA传递到PpcA,最终由外膜蛋白OmcB、OmcE、OmcS及OmcZ接受电子,并在Type Ⅳ pili的共同作用下将电子传递到胞外电子受体。本文最后指出目前对Shewanella与Geobacter胞内电子转移研究尚不清楚的地方提出展望。     

微生物铁呼吸机制研究进展

铁呼吸是厌氧环境中普遍存在的一种微生物代谢形式,多种古生菌和细菌都能进行铁呼吸.Fe(Ⅲ)的地球化学丰度比较高,为Fe(Ⅲ)还原菌提供了充足的电子受体,但自然中Fe(Ⅲ)多以不溶形式存在,使电子传递受阻.本文介绍了Fe(Ⅲ)还原菌的多样性,总结了4种铁呼吸机制:直接接触机制、螯合促溶机制、电子穿梭机制、纳米导线辅助机制,并对铁呼吸机制未来的研究方向进行了展望.     

Evidence for the coexistence of direct and riboflavin-mediated interspecies electron transfer in Geobacter co-culture

Geobacter species can secrete free redox-active flavins, but the role of these flavins in the interspecies electron transfer (IET) of Geobacter direct interspecies electron transfer (DIET) co-culture is unknown. Here, we report the presence of a new riboflavin-mediated interspecies electron transfer (RMIET) process in a traditional Geobacter DIET co-culture; in this process, riboflavin contributes to IET by acting as a free-form electron shuttle between free Geobacter species and serving as a bound cofactor of some cytochromes in Geobacter co-culture aggregates. Multiple lines of evidence indicate that RMIET facilitates the primary initiation of syntrophic growth between Geobacter species before establishing the DIET co-culture and provides additional ways alongside the DIET to transfer electrons to achieve electric syntrophy between Geobacter species. Redox kinetic analysis of riboflavin on either Geobacter species demonstrated that the Gmet_2896 cytochrome acts as the key riboflavin reduction site, while riboflavin oxidation by Geobacter sulfurreducens is the rate-limiting step in RMIET, and the RMIET makes only a minor contribution to IET in Geobacter DIET co-culture. The discovery of a new RMIET process in Geobacter DIET co-culture suggests the complexity of IET in syntrophic bacterial communities and provides suggestions for the careful examination of the IET of other syntrophic co-cultures.     

微生物种间直接电子传递研究进展

一直以来氢气和甲酸被认为是微生物间电子传递的中间电子传递体。近年来的研究发现,微生物之间可以通过种间直接电子传递(DIET)来替代氢气/甲酸传递。DIET作为一种新发现的微生物间电子传递途径,其电子传递效率要高于传统的种间氢气/甲酸传递。DIET这一新发现改变了微生物互营生长代谢必须依赖氢气或甲酸等电子载体的传统认识,为今后研究微生物互营现象打开了新视角。虽然DIET研究取得了很大进展,但是目前对能够进行DIET的微生物种类、DIET机制及影响DIET的因素尚缺乏深入研究。本文首先概述了能形成DIET的微生物,然后重点分析了能够进行DIET的电子供体微生物胞外电子传递的机制和电子受体微生物直接利用胞外电子的分子机制,最后阐述了导电材料对DIET的影响,并提出了DIET今后的研究方向,旨在为DIET研究提供参考。     

Carbon nanotubes accelerate methane production in pure cultures of methanogens and in a syntrophic coculture

Carbon materials have been reported to facilitate direct interspecies electron transfer (DIET) between bacteria and methanogens improving methane production in anaerobic processes. In this work, the effect of increasing concentrations of carbon nanotubes (CNT) on the activity of pure cultures of methanogens and on typical fatty acid‐degrading syntrophic methanogenic coculture was evaluated. CNT affected methane production by methanogenic cultures, although acceleration was higher for hydrogenotrophic methanogens than for acetoclastic methanogens or syntrophic coculture. Interestingly, the initial methane production rate (IMPR) by Methanobacterium formicicum cultures increased 17 times with 5 g·L^?1 CNT. Butyrate conversion to methane by Syntrophomonas wolfei and Methanospirillum hungatei was enhanced (～1.5 times) in the presence of CNT (5 g·L^?1), but indications of DIET were not obtained. Increasing CNT concentrations resulted in more negative redox potentials in the anaerobic microcosms. Remarkably, without a reducing agent but in the presence of CNT, the IMPR was higher than in incubations with reducing agent. No growth was observed without reducing agent and without CNT. This finding is important to re‐frame discussions and re‐interpret data on the role of conductive materials as mediators of DIET in anaerobic communities. It also opens new challenges to improve methane production in engineered methanogenic processes.     

Metabolism of sulfate-reducing prokaryotes

Dissimilatory sulfate reduction is carried out by a heterogeneous group of bacteria and archaea that occur in environments with temperatures up to 105 °C. As a group together they have the capacity to metabolize a wide variety of compounds ranging from hydrogen via typical organic fermentation products to hexadecane, toluene, and several types of substituted aromatics. Without exception all sulfate reducers activate sulfate to APS; the natural electron donor(s) for the ensuing APS reductase reaction is not known. The same is true for the reduction of the product bisulfite; in addition there is still some uncertainty as to whether the pathway to sulfide is a direct six-electron reduction of bisulfite or whether it involves trithionate and thiosulfate as intermediates. The study of the degradation pathways of organic substrates by sulfate-reducing prokaryotes has led to the discovery of novel non-cyclic pathways for the oxidation of the acetyl moiety of acetyl-CoA to CO₂. The most detailed knowledge is available on the metabolism ofDesulfovibrio strains, both on the pathways and enzymes involved in substrate degradation and on electron transfer components and terminal reductases. Problems encountered in elucidating the flow of reducing equivalents and energy transduction are the cytoplasmic localization of the terminal reductases and uncertainties about the electron donors for the reactions catalyzed by these enzymes. New developments in the study of the metabolism of sulfate-reducing bacteria and archaea are reviewed.     

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.     

Microbial extracellular electron transfer and its relevance to iron corrosion

Extracellular electron transfer (EET) is a microbial metabolism that enables efficient electron transfer between microbial cells and extracellular solid materials. Microorganisms harbouring EET abilities have received considerable attention for their various biotechnological applications, including bioleaching and bioelectrochemical systems. On the other hand, recent research revealed that microbial EET potentially induces corrosion of iron structures. It has been well known that corrosion of iron occurring under anoxic conditions is mostly caused by microbial activities, which is termed as microbiologically influenced corrosion (MIC). Among diverse MIC mechanisms, microbial EET activity that enhances corrosion via direct uptake of electrons from metallic iron, specifically termed as electrical MIC (EMIC), has been regarded as one of the major causative factors. The EMIC‐inducing microorganisms initially identified were certain sulfate‐reducing bacteria and methanogenic archaea isolated from marine environments. Subsequently, abilities to induce EMIC were also demonstrated in diverse anaerobic microorganisms in freshwater environments and oil fields, including acetogenic bacteria and nitrate‐reducing bacteria. Abilities of EET and EMIC are now regarded as microbial traits more widespread among diverse microbial clades than was thought previously. In this review, basic understandings of microbial EET and recent progresses in the EMIC research are introduced.     

Exocellular electron transfer in anaerobic microbial communities

Exocellular electron transfer plays an important role in anaerobic microbial communities that degrade organic matter. Interspecies hydrogen transfer between microorganisms is the driving force for complete biodegradation in methanogenic environments. Many organic compounds are degraded by obligatory syntrophic consortia of proton-reducing acetogenic bacteria and hydrogen-consuming methanogenic archaea. Anaerobic microorganisms that use insoluble electron acceptors for growth, such as iron- and manganese-oxide as well as inert graphite electrodes in microbial fuel cells, also transfer electrons exocellularly. Soluble compounds, like humic substances, quinones, phenazines and riboflavin, can function as exocellular electron mediators enhancing this type of anaerobic respiration. However, direct electron transfer by cell-cell contact is important as well. This review addresses the mechanisms of exocellular electron transfer in anaerobic microbial communities. There are fundamental differences but also similarities between electron transfer to another microorganism or to an insoluble electron acceptor. The physical separation of the electron donor and electron acceptor metabolism allows energy conservation in compounds as methane and hydrogen or as electricity. Furthermore, this separation is essential in the donation or acceptance of electrons in some environmental technological processes, e.g. soil remediation, wastewater purification and corrosion.     

Advances in direct interspecies electron transfer and conductive materials: Electron flux,organic degradation and microbial interaction

Direct interspecies electron transfer (DIET) via electrically conductive pili (e-pili) and c-type cytochrome between acetogens and methanogens has been proposed as an essential pathway for methane production. Supplements of conductive materials have been extensively found to promote methane production in microbial anaerobic treatment systems. This review comprehensively presents recent findings of DIET and the addition of conductive materials for methanogenesis and summarizes important results through aspects of electron flux, organic degradation, and microbial interaction. Conductive materials improve DIET and methanogenesis by acting as either substitute of e-pili or electron conduit between e-pili and electron acceptors. Other effects of conductive materials such as the change of redox potential may also be important factors for the stimulation. The type and organic loading rate of substrates affect the occurrence of DIET and stimulating effects of conductive materials. Geobacter, which can participate in DIET, were less enriched in anaerobic systems cultivated with non-ethanol substrates, suggesting the existence of other syntrophs with the capability of DIET. The coupling of communication systems such as quorum sensing may be a good strategy to achieve the formation of biofilm or granule enriched with syntrophic partners capable of DIET.     

微生物种间直接电子传递机理及应用研究进展

微生物胞内产生的电子转移到其他电子受体而获得能量的过程称为微生物胞外电子传递,其中,另一微生物作为电子受体时发生的电子传递称为微生物种间电子传递。根据微生物种间电子传递机制,可分间接种间电子传递和种间直接电子传递。由于种间直接电子传递不需要其他物质介导,因此较间接种间电子传递效率更高、能量利用更高。本文系统阐述了微生物进行胞外电子传递的机理及应用,重点分析了种间直接电子传递机理,并概述种间直接电子传递应用领域,为寻找更多电连接的微生物群落以及应用微生物提供参考。