石油烃厌氧生物降解代谢产物研究进展

石油烃厌氧生物降解代谢产物的分析对于石油烃厌氧降解机制的研究、功能微生物的筛选以及微生物活动的原位监测具有指示性作用.综述了近年来石油烃厌氧生物降解代谢产物的研究进展.石油烃厌氧降解的初始活化方式主要包括脱氢羟基化、加延胡索酸以及羧化等.其中,加延胡索酸是不同种类的微生物通常采用的代谢方式.同时,将代谢产物按照气体、无机离子和有机酸进行分类,并针对各类物质特别是瞬时性、低浓度的有机酸类产物常采用的分析方法进行归纳.通过实例强调了代谢产物作为潜在生物标记物的应用,并对石油烃厌氧降解代谢产物分析方法的发展提出展望.图3参58     

稳定同位素技术在污染环境生物修复研究中的应用

稳定同位素技术主要应用于地球化学,它是将人工合成的同位素标记特定的化合物,追踪标记物在生命活动中的变化规律,目前该项技术也广泛应用于环境微生物学、生态学、生物医学等领域.生物修复是利用存在于土壤、地下水和海洋等环境中的生物特别是微生物将有毒、有害的污染物降解为二氧化碳和水,或转化为无害物质,从而使污染的生态环境修复为正常生态环境的过程.这些降解微生物都来自于小部份可培养微生物,对于大部份未可培养降解微生物,通常在实验室条件下很难得到.而利用稳定同位素技术,如13C标记底物,收集利用该底物的微生物核酸,就可以得到具有降解作用的功能微生物,为环境污染生物修复提供重要的菌源和功能基因.环境中的许多物质都可以用SIP来标记,这些标记物主要有PLFA-SIP、DNA-SIP、RNA-SIP等,它们都可以用来在复杂样本中进行有特殊代谢功能微生物的鉴定和分析,在利用微生物进行生物修复中具有重要的意义.图2表1参42     

石油烃降解酶及其基因的结构、功能和表达调控

研究烃降解酶及其基因是进行石油微生物分子检测和工程菌构建的重要基础.本文对目前烃降解酶及其基因的结构、功能和调控机制的最新研究进展进行了总结.催化烷烃好氧降解的起始酶有几类加氧酶,膜整合甲烷单加氧酶、萘-1,2-双加氧酶和异丙苯双加氧酶的晶体结构已经被解析.烷基或芳基琥珀酸合酶催化烃厌氧代谢的主要起始反应,而Azoarcus sp.乙苯厌氧代谢起始反应由乙苯脱氢酶催化.在细菌中,烃代谢相关基因主要通过形成操纵子进行表达调控,基因转录受烃或类似物诱导,并受细胞全局调控.一些微生物由于存在多种烃代谢途径而可能具有复杂的基因调控机制.此外,生态学研究表明,环境中烃降解基因的诱导动态与实验室内纯培养分析不同.在分析石油降解工程菌构建有待解决问题的基础上,提出了烃代谢综合调控和环境中相关酶及基因诱导研究的重要性,并对未来烃降解酶及其基因在有毒物降解理论研究和生物修复上的应用进行了展望.     

地下环境中铁氧化物生物异化还原耦合降解硝基苯的影响因素研究

通过静态实验模拟地下水厌氧环境,研究了碳源、电子受体、初始微生物量和硝基苯浓度等因素对地下环境中微生物异化铁还原作用耦合降解硝基苯的影响。实验选取乙酸铵、葡萄糖、柠檬酸铵为碳源,针铁矿、赤铁矿为电子受体,进行异化铁还原协同耦合降解硝基苯。结果表明：乙酸铵为碳源时硝基苯的降解率最高为78.5%;针铁矿作电子受体时硝基苯降解效果比赤铁矿好,且0.3 mg·L-1效果最好;微生物量对耦合体系中硝基苯降解率有影响但并不显著,最适宜的初始微生物量为4×108cells·mL-1;硝基苯质量浓度在50～150 mg·L-1时,硝基苯浓度与降解率呈负相关,其中对50 mg·L-1的初始硝基苯质量浓度降解效果最好。因此,碳源和电子受体对地下环境中微生物异化铁还原作用耦合降解硝基苯具有重要的影响。     

微生物直接电子传递:甲烷代谢古菌研究进展

甲烷是重要的温室气体,同时也是广泛的可再生能源.深刻认识甲烷代谢过程中的微观机理可为人类实现甲烷的减排及其作为能源的合理利用打下坚实的理论基础.古菌介导的直接电子传递(DET)作为甲烷代谢的重要途径,已成为近年来环境微生物领域的研究热点.本文对互营氧化产甲烷、电能无机自养产甲烷以及厌氧甲烷氧化3个过程中参与DET的微生物进行综述,并着重阐述它们各自的发生机理. DET既存在于甲烷合成代谢,又涉及厌氧甲烷氧化.前者根据电子来源的不同,可分为微生物种间DET产甲烷和电能无机自养产甲烷两种类型.后者则是甲烷氧化古菌将甲烷氧化产生的电子传递至胞外电子受体.在甲烷合成代谢过程中,产甲烷古菌主要通过互营细菌外膜细胞色素蛋白、菌毛或导电性固体3种方式进行电子的直接吸收.相反,甲烷氧化古菌可通过外膜细胞色素蛋白将电子传递至胞外固体或微生物.今后对于古菌介导的DET研究将集中在甲烷代谢电子传递链的各个组成部分及其与细菌之间的相互作用,以便将DET机制用于实际问题的解决.     

细菌偶氮还原研究进展

细菌偶氮还原是在细菌偶氮还原酶作用下偶氮复合物分解为芳香氨的过程,多种细菌都具有偶氮还原功能.有氧条件下细菌的偶氮还原是在特异性的偶氮还原酶作用下完成的,对氧气不敏感.而厌氧菌和兼性厌氧菌的偶氮还原是在厌氧或兼性厌氧条件下由一些氧化还原中间体作为电子穿梭体与偶氮复合物作用的非特性的还原过程.电子穿梭体与偶氮复合物作用是一个纯化学氧化还原过程,其中偶氮复合物作为电子受体接受电子.厌氧偶氮还原是细菌偶氮还原的主要形式.偶氮还原在厌氧-好氧染料废水处理系统和偶氮水溶胶聚合物作为结肠靶向给药载体的研究中有重要应用.图2表1参45     

壬基酚在海河沉积物中的耗氧和厌氧降解

研究了壬基酚（NP）在海河沉积物中的降解.结果表明,NP在沉积物的耗氧降解分为快速和慢速降解阶段,半衰期分别为3.20-9.87d和21.66-385.11d.NP在沉积物中的厌氧降解缓慢,降解半衰期为160.65-203.88d.加入电子受体NaNO3和Na2SO4促进了NP的厌氧降解,并且NaNO3促进作用较大.当沉积物中NP降低到一定浓度以后就很难继续降解,说明沉积物中一些活性吸附点位对NP的锁定作用降低了其生物有效性.盐度不利于NP的耗氧和厌氧降解.     

石油烃厌氧生物降解研究进展

烃是由碳和氢两种元素组成的稳定化合物,长期以来人们普遍认为烃的生物降解只能在好氧条件下进行.近年来国外的研究表明,厌氧微生物能以硝酸盐、硫酸盐、三价铁或二氧化碳等为电子受体代谢烃,烃厌氧代谢的初始反应机理主要有延胡索酸盐结合反应、羧化反应、羟基化反应和甲基化反应,但国内还未见相关报道.本文综述了烃厌氧降解微生物及其代谢机理,并以甲苯为例概述了烃厌氧生物降解过程中的起始反应酶和作用机制.结合国外烃厌氧降解的研究进展和本实验室的工作,作者也提出了自己的思考和看法.     

一株2,2’,4,4’-四溴联苯醚(BDE-47)厌氧降解菌的筛选鉴定及降解特性

为去除环境中BDE-47的残留,通过以BDE-47为碳源的选择性培养基驯化,从电子垃圾拆解厂的土壤中分离出了1株厌氧降解BDE-47的纯菌种,命名为XM,并研究其对BDE-47的降解特性.经16S rDNA鉴定,XM属于兼性肠杆菌(Enterobacter sp.),当BDE-47浓度为525μg/L,初始接菌量为7.1×10~5 cells/m L时,培养35 d后降解率为35.8%,降解产物中检测到BDE-28.BDE-47的降解反应符合一级动力学,拟合结果为ln C_t=-0.104t+6.22.选择以铁离子、硝酸根和硫酸根作为降解过程中外加的电子受体,BDE-47的降解率明显提高,分别为49.8%、59.1%和67.3%.以上研究结果表明,菌株XM能够有效地降解BDE-47,在电子垃圾污染的生物修复方面具有较好的参考和应用价值.     

产甲烷古菌与电子传递体相互作用机制研究进展

厌氧消化是实现有机废弃物资源化最有效的技术之一，实现形式是产生生物沼气.作为一种清洁能源，生物沼气可以有效减少化石燃料的使用，进而减少温室气体的排放.产甲烷古菌位于厌氧发酵链末端，是生物沼气主要成分甲烷的直接生产者.在厌氧消化系统中，产甲烷古菌与发酵链前端微生物以及各种天然和人工电子传递体存在着活跃的电子互营过程，对于维持厌氧消化系统的稳定性和改善生物沼气的生成效率具有重要作用.本文综述近年来报道的在强化厌氧消化过程中常用的铁基与碳基电子传递体与产甲烷古菌的相互作用机制，着重介绍两类电子传递体通过自身氧化还原反应或物理性质与产甲烷古菌细胞膜上的氢酶和细胞色素c进行电子互营的微观作用机理，分析两类电子传递体通过参与胞外电子传递过程与产甲烷古菌能量代谢可能存在的耦合机制，其中乙酸型产甲烷古菌基于电子歧化传递在进行胞外三价铁呼吸过程中存储能量，从而增强产甲烷代谢，改变了目前对甲烷生成的生化和生态学理解，极大推进了产甲烷古菌与胞外电子传递体相互作用的研究.产甲烷古菌胞外电子传递路径的不清晰和其细胞膜上蛋白功能的不确定是制约产甲烷古菌与电子传递体相互作用机制研究的重要因素.因此提出利用快速发展的...     

影响厌氧氨氧化与甲烷化反硝化耦合的因素

氨氮、氮氧化物对产甲烷菌有一定的抑制作用,但可以通过驯化去除毒性.亚硝酸盐在厌氧氨氧化菌作用下与氨发生厌氧氨氧化反应.虽然厌氧氨氧化菌是自养菌,但具有异养代谢能力,并且NO2可提高厌氧氨氧化菌的活性.因此,通过特殊的反应器技术,将厌氧氨氧化菌与甲烷菌、反硝化菌复合在一个有利的微生态环境中,充分发挥它们之间的协同耦合作用,把有机物转化为清洁能源又同时脱氮,是极有前景的废水厌氧(缺氧)处理研究新方向.表1参31     

Anaerobic biodegradation of trimethoprim with sulfate as an electron acceptor

Anaerobic biodegradation of trimethoprim (TMP) coupled with sulfate reduction. Demethylation of TMP is the first step in the acclimated microbial consortia. The potential degraders and fermenters were enriched in the acclimated consortia. Activated sludge and river sediment had similar core microbiomes. Trimethoprim (TMP) is an antibiotic frequently detected in various environments. Microorganisms are the main drivers of emerging antibiotic contaminant degradation in the environment. However, the feasibility and stability of the anaerobic biodegradation of TMP with sulfate as an electron acceptor remain poorly understood. Here, TMP-degrading microbial consortia were successfully enriched from municipal activated sludge (AS) and river sediment (RS) as the initial inoculums. The acclimated consortia were capable of transforming TMP through demethylation, and the hydroxyl-substituted demethylated product (4-desmethyl-TMP) was further degraded. The biodegradation of TMP followed a 3-parameter sigmoid kinetic model. The potential degraders (Acetobacterium, Desulfovibrio, Desulfobulbus, and unidentified Peptococcaceae) and fermenters (Lentimicrobium and Petrimonas) were significantly enriched in the acclimated consortia. The AS- and RS-acclimated TMP-degrading consortia had similar core microbiomes. The anaerobic biodegradation of TMP could be coupled with sulfate respiration, which gives new insights into the antibiotic fate in real environments and provides a new route for the bioremediation of antibiotic-contaminated environments.     

电子穿梭体及其介导的环境与地球化学过程研究进展

电子穿梭体是一类可通过自身氧化还原介导电子转移的化学物质的统称.在地球表层系统中,电子穿梭体可加速微生物向细胞外部进行的电子传递,参与矿物的微生物还原,驱动碳、氮、硫元素循环,并偶联有机污染物降解和重金属迁移转化.电子穿梭在自然界中存在广泛,对于元素循环、污染物环境行为以及微生物的生存行为影响深远,具有重要的环境地球化...     

Anaerobic phenanthrene biodegradation with four kinds of electron acceptors enriched from the same mixed inoculum and exploration of metabolic pathways

Anaerobic phenanthrene biodegradation enriched process was described in detail. The enriched bacterial communities were characterized under four redox conditions. The enriched archaeal communities were stated under high percentage conditions. Relatively intact pathways of anaerobic phenanthrene biodegradation were proposed. Polycyclic aromatic hydrocarbons (PAHs) are widespread and persistent contaminants worldwide, especially in environments devoid of molecular oxygen. For lack of molecular oxygen, researchers enhanced anaerobic zones PAHs biodegradation by adding sulfate, bicarbonate, nitrate, and iron. However, microbial community reports of them were limited, and information of metabolites was poor except two-ring PAH, naphthalene. Here, we reported on four phenanthrene-degrading enrichment cultures with sulfate, bicarbonate, nitrate, and iron as electron acceptors from the same initial inoculum. The high-to-low order of the anaerobic phenanthrene biodegradation rate was the nitrate-reducing conditions>sulfate-reducing conditions>methanogenic conditions>iron-reducing conditions. The dominant bacteria populations were Desulfobacteraceae, Anaerolinaceae, and Thermodesulfobiaceae under sulfate-reducing conditions; Moraxellaceae, Clostridiaceae, and Comamonadaceae under methanogenic conditions; Rhodobacteraceae, Planococcaceae, and Xanthomonadaceae under nitrate-reducing conditions; and Geobacteraceae, Carnobacteriaceae, and Anaerolinaceae under iron-reducing conditions, respectively. Principal component analysis (PCA) indicated that bacteria populations of longtime enriched cultures with four electron acceptors all obtained significant changes from original inoculum, and bacterial communities were similar under nitrate-reducing and iron-reducing conditions. Archaea accounted for a high percentage under iron-reducing and methanogenic conditions, and Methanosarcinaceae and Methanobacteriaceae, as well as Methanobacteriaceae, were the dominant archaea populations under iron-reducing and methanogenic conditions. The key steps of phenanthrene biodegradation under four reducing conditions were carboxylation, further ring system reduction, and ring cleavage.     

Biological conversion pathways of sulfate reduction ammonium oxidation in anammox consortia

• The SRAO phenomena tended to occur only under certain conditions. • High amount of biomass and non-anaerobic condition is requirement for SRAO. • Anammox bacteria cannot oxidize ammonium with sulfate as electron acceptor. • AOB and AnAOB are mainly responsible for ammonium conversion. • Heterotrophic sulfate reduction mainly contributed to sulfate conversion. For over two decades, sulfate reduction with ammonium oxidation (SRAO) had been reported from laboratory experiments. SRAO was considered an autotrophic process mediated by anammox bacteria, in which ammonium as electron donor was oxidized by the electron acceptor sulfate. This process had been attributed to observed transformations of nitrogenous and sulfurous compounds in natural environments. Results obtained differed largely for the conversion mole ratios (ammonium/sulfate), and even the intermediate and final products of sulfate reduction. Thus, the hypothesis of biological conversion pathways of ammonium and sulfate in anammox consortia is implausible. In this study, continuous reactor experiments (with working volume of 3.8L) and batch tests were conducted under normal anaerobic (0.2≤DO<0.5 mg/L) / strict anaerobic (DO<0.2 mg/L) conditions with different biomass proportions to verify the SRAO phenomena and identify possible pathways behind substrate conversion. Key findings were that SRAO occurred only in cases of high amounts of inoculant biomass under normal anaerobic condition, while absent under strict anaerobic conditions for same anammox consortia. Mass balance and stoichiometry were checked based on experimental results and the thermodynamics proposed by previous studies were critically discussed. Thus anammox bacteria do not possess the ability to oxidize ammonium with sulfate as electron acceptor and the assumed SRAO could, in fact, be a combination of aerobic ammonium oxidation, anammox and heterotrophic sulfate reduction processes.     

混合菌群生物燃料电池的产电机理与特性

利用厌氧污泥为接种源构建双室微生物燃料电池(Microbial fuel cell,MFC),研究其电子传递机制,并考察其底物利用谱及阴极电子受体对产电性能的影响.结果表明:该MFC主要通过生物膜机制实现电子从有机物到固体电极的传递过程.该混合菌MFC的底物利用谱范围广泛,单糖、二糖、小分子有机酸等有机物均可作为电子供体产电,其中以蔗糖和乳糖为底物产电效果较好,最大功率密度分别为69.69 mW/m2和60.75 mW/m2;而以乙醇为底物时,COD负荷最高,达123.55 mg L-1d-1.阴极不同电子受体对混合菌群MFC的产电性能也有显著影响,其中以KMnO4为电子受体电池性能最好,最大功率密度达1 396.74 mW/m2.     

水稻土中铁的厌氧生物氧化还原循环对砷的影响（The Effect of Anaerobic Redox Cycling of Iron on Arsenic Mobility in Paddy）

应用砷污染水稻土的厌氧富集培养,探讨水稻土中潜在存在铁厌氧生物循环及其对氮和砷的耦合作用.富集培养直接证明了水稻土中铁的厌氧生物循环:三价铁(人工合成针铁矿)在厌氧条件下被逐渐还原成二价铁;铁还原过程结束并外源添加硝酸根时,培养基中新生的二价铁在依赖于硝酸根的铁氧化菌的作用下被氧化;当提供新的电子供体乙酸时,生物合成的铁矿重新被还原.在铁氧化还原循环过程中,随着铁的还原,培养基中砷的浓度不断增加,反之,当铁逐渐氧化的同时不断地吸附固定培养基中的砷.在铁的厌氧氧化阶段,铁氧化的同时硝酸根被还原,培养基中积累了NH4+和NO2-.因此,厌氧水稻土中可以进行完整的铁氧化还原循环,同时这个循环过程耦合了氮和砷的迁移转化.     

Assessing biodegradation benefits from dispersal networks

The performance of biodegradation of organic pollutants in soil often depends on abiotic conditions and the bioavailability of these pollutants to degrading bacteria. In this context, bacterial dispersal is an essential aspect. Recent studies on the potential promotion of bacterial dispersal by fungal hyphae raised the idea of specifically applying fungal networks to accelerate bacterial degradation processes in situ. Our objective is to investigate these processes and their performance via simulation modelling and address the following questions: (1) Under what abiotic conditions can dispersal networks significantly improve bacterial degradation? and (2) To what extent does the spatial configuration of the networks influence the degradation performance? To answer these questions, we developed a spatially explicit bacterial colony model, which is applied to controlled laboratory experiments with Pseudomonas putida G7 organisms as a case study. Using this model, we analyzed degradation performance in response to different environmental scenarios and showed that conditions of limited bacterial dispersal also limit degradation performance. Under such conditions, dispersal networks have the highest potential for improving the bioavailability of pollutants to bacteria. We also found that degradation performance significantly varies with the spatial configuration of the dispersal networks applied and the time horizon over which performance is assessed. Regarding future practical applications, our results suggest that (1) fungal networks may dramatically improve initially adverse conditions for biodegradation of pollutants in soil, and (2) the network's spatial structure and accessibility are decisive for the success of such tasks.     

轮虫在生态毒理学中的研究进展

轮虫是浮游动物中的重要类群,因具有生活周期短、对毒物的敏感性高、有两性生殖方式、易培养和实用性强等优势特征,已被用作生态毒理学实验中重要的模式生物,颇有研究前景。本文简介了轮虫作为受试生物的特点及其生活史,并从实验方法、污染物种类、测试指标等角度对近几十年来轮虫在生态毒理学中的应用现状、主要成果和最新进展作了概述,主要包括轮虫在常规毒性评价中的应用:急性毒性、慢性毒性以及对其行为的影响效应;综述了几类典型环境污染物,例如重金属、UV辐射、持久性有机污染物以及农药化合物的胁迫对轮虫的毒性效应,特别对目前存在的问题、研究热点及今后需要加强研究的方向进行了讨论和展望,以期为应用轮虫生态毒理学进行环境污染及风险评估提供参考。     

厌氧发酵过程pH对微生物多样性和产物分布的影响

采用PCR-DGGE技术,研究了不同pH条件下蔬菜类有机垃圾厌氧发酵过程中的微生物多样性,探讨了微生物群落结构与发酵产物分布的关系.Shannon指数分析表明,pH=7和pH=8时的微生物多样性较高,随时间变化规律相似,而pH=5时的微生物多样性较低.UPGMA聚类分析和PCA分析结果也表明,pH=7和pH=8时的微生物群落结构相似,pH=5与之显著不同.在不同pH条件下的优势菌属都是乳酸细菌和梭菌。微生物多样性的变化与发酵产物分布具有一定相关性,pH通过影响微生物群落结构的变化最终影响发酵产物的分布.图4表2参27