多氯联苯微生物脱氯研究进展

多氯联苯(polychlorinated biphenyls,PCBs)是环境中典型的氯代持久性有机污染物.微生物脱氯是一种氯代有机物自然降解模式,对全球PCBs特别是高氯代同系物消减起到至关重要的作用.厌氧条件下高氯代PCBs能够发生脱氯反应,使其毒性大大降低,脱氯后形成的低氯代化合物可以进一步好氧降解,直至完全矿化.本文综述了PCBs生物脱氯的研究进展,介绍了微生物脱氯反应的机理和特征、参与微生物脱氯过程的专性脱氯菌等,探讨了该微生物过程的影响因素及厌氧脱氯与好氧降解耦合的意义,并对脱氯微生物群落的复杂代谢网络研究、专性脱氯新菌种筛选及其污染地实际修复应用等未来研究方向进行了展望.     

纳米零价铁耦合假单胞菌协同高效降解五氯苯

【背景】氯苯类化合物广泛应用于工业化合物的生产,由于其具有高毒性和环境持久性的特点,对人类健康和生态环境造成严重威胁,寻找高效降解这类化合物的新方法成为研究热点。【目的】将纳米零价铁与假单胞菌耦合,探究其在好氧条件下对五氯苯的降解效果和降解机理。【方法】建立纳米零价铁和假单胞菌降解五氯苯的反应体系,通过测定各反应体系中五氯苯及其中间产物的浓度,以及观测细菌的生长状况变化等,分析反应体系的降解效果及其可能的降解机理。【结果】纳米零价铁耦合假单胞菌的反应体系相对于两者的单一体系表现出更好的降解效果,36h的降解率可达55.4%,反应过程符合伪一级反应动力学,速率常数为0.02048h~(-1)。根据GC-MS所测的中间产物推测,该体系的反应机理为纳米零价铁在好氧条件下反应产生羟基自由基,攻击五氯苯使其变为低氯苯类化合物,假单胞菌进一步利用这些低氯苯类化合物;同时,假单胞菌又为纳米零价铁提供附着位点,有效地降低了纳米零价铁的聚集性,提高了反应活性。【结论】研究建立的纳米零价铁耦合假单胞菌反应体系对五氯苯具有较好的降解效果,为含有高氯代苯类等有机污染物的复杂环境的修复提供参考。     

零价铁对2,4-二氯酚生物还原脱氯的影响研究

采用间歇试验,接种驯化两月的厌氧混合微生物,考察厌氧体系中添加零价铁（Fe^0）对2,4-二氯酚（2,4-DCP）生物还原脱氯效果的影响,并对影响“Fe^O＋微生物”体系的一些因素进行了探索。结果显示：与零价铁或微生物的单独作用相比,“Fe^O＋微生物”体系能够有效促进2,4-DCP的脱氯反应,最佳Fe^O投加量和微生物接种量分别为0.5g／L和376.2mgVSS／L;初始pH=8.0对2,4-DCP的转化效果最好,偏酸性环境不利于污染物转化;微生物接种量与铁用量之间有一适宜比例,一定范围内增加微生物接种量可催生出更多可降解污染物的酶或酶系,提高2,4-DCP的降解效果。     

零价铁对2,4-二氯酚生物还原脱氯的影响研究

采用间歇试验, 接种驯化两月的厌氧混合微生物, 考察厌氧体系中添加零价铁(Fe0)对2,4-二氯酚(2,4-DCP)生物还原脱氯效果的影响, 并对影响“Fe0+微生物”体系的一些因素进行了探索。结果显示：与零价铁或微生物的单独作用相比, “Fe0+微生物”体系能够有效促进2,4-DCP的脱氯反应, 最佳Fe0投加量和微生物接种量分别为0.5 g/L和376.2 mgVSS/L; 初始pH = 8.0对2,4-DCP的转化效果最好, 偏酸性环境不利于污染物转化; 微生物接种量与铁用量之间有一适宜比例, 一定范围内增加微生物接种量可催生出更多可降解污染物的酶或酶系, 提高2,4-DCP的降解效果。     

植物修复多氯联苯研究进展

综述了植物修复持久性有机污染物多氯联苯（PCBs）的研究进展,重点阐述了植物对PCBs的去除作用和机理,植物在从环境中去除PCBs的过程中,不仅仅是作为微生物降解的支持者,而且还作为积极的参与者对PCBs进行代谢：一方面植物通过根系从环境中吸收和积累PCBs,并将吸收的PCBs转化为非毒性的代谢产物累积于植物组织中;另一方面植物释放促进PCBs降解的酶直接降解PCBs,或释放根系分泌物,增加根际微生物的数量,提高其活性间接降解PCBs.文中对植物修复PCBs的影响因素如植物组织培养的类型、生物量、PCBs的初始浓度以及PCBs的类型、理化性质等进行了讨论.     

硝酸盐型厌氧铁氧化菌的种类、分布和特性

硝酸盐型厌氧铁氧化(NAFO)是指微生物在厌氧条件下利用硝酸盐或亚硝酸盐作为电子受体,将低价铁(二价铁或零价铁)氧化为高价铁(三价铁)的过程。具有NAFO代谢能力的微生物称为硝酸盐型厌氧铁氧化菌(NAFOM)。NAFO是微生物领域的重大发现,也是环境领域开发新型脱氮技术和地学领域研究铁、氮循环的理论依据。整理文献报道的NAFOM资料,分析NAFOM系统发育性状,探讨典型NAFOM的生态分布及其营养、代谢特性,以期为NAFOM菌种资源的开发、地球铁素和氮素循环的研究、NAFO过程的优化提供借鉴。     

土壤和沉积物中多氯代有机化合物厌氧降解研究进展

多氯代有机化合物(PCOCs)是土壤和沉积物中的典型污染物,厌氧条件下PCOCs能够发生脱氯发应,从而使其毒性大大降低,脱氯后形成的低氯代化合物可以进一步好氧降解,直至完全矿化。从PCOCs的降解过程出发,重点综述了几种典型PCOCs的厌氧脱氯机理以及几种重要影响因素;阐明了脱氯反应是PCOCs厌氧降解的关键步骤,反应的发生必须有还原剂提供电子,微生物的参与尤为重要;同时展望了同位素示踪法在研究PCOCs降解机制上的应用,以及开发高效降解PCOCs微生物的必要性等。     

铁还原菌在水资源再生与能源转化领域的研究进展

铁还原菌是一种典型的异化金属还原菌,广泛分布于海洋沉积物、陆地深地层等自然环境,该类细菌可以将铁氧化物中的Fe(Ⅲ)还原为Fe(Ⅱ),在铁、碳的生物地球化学铁循环中发挥重要作用。铁还原菌的末端电子不局限于Fe(Ⅲ),还可以是其他高价金属、有机污染物,可用于土壤、地下水的污染修复和毒性削减。在微生物电化学系统中,铁还原菌氧化有机物产生的电子直接传递给电极,可以产生电能。基于这种独特的胞外电子传递方式,衍生出了微生物燃料电池、微生物电解池、微生物脱盐电池、微生物燃料电池耦合芬顿反应以及光催化微生物燃料电池,常用于微生物发电、生物传感器、生物制氢、定向发酵、海水淡化、生物脱盐和污染物分解矿化。本文从异化铁还原菌的代谢机制、微生态作用、环境修复、水资源再生与能源转化四个方面,综述了铁还原菌的作用原理及国内外研究现状,分析论述了目前亟需解决的关键问题和未来的研究方向,以期为铁还原菌的基础理论研究和应用技术研发提供参考。     

微生物降解多氯联苯的研究进展

多氯联苯是一种持续性有机污染物,在自然环境中很难降解。在目前研究的降解方法中,微生物降解最具潜力。本文对多氯联苯微生物降解的研究进展进行了综述,包括厌氧还原脱氯,好氧氧化以及生物表面活性剂的作用,介绍了几种降解方法耦合应用的现状和前景,指出了应用中存在的问题和今后的发展方向。     

蔷薇类离体培养物降解多氯联苯

多氯联苯类(PCBs)物质是毒性很强的环境污染物。PCBs由于它的广泛存在、在被污染的食物链高层次上的富集以及在普通环境条件下的抗降解性而受到特别重视。已经证明,在植物-水-土壤系统中,PCBs可以被降解,但还不清楚PCBs在这类系统中的降解作用是由植物还是土壤微生物或两者共同代谢掉的。美国俄克拉何马大学的J.Fletcher及其同事和美国环境保护署已经证明:无菌条件下     

Microbial reductive dehalogenation of polychlorinated biphenyls

Under anaerobic conditions, microbial reductive dechlorination of polychlorinated biphenyls (PCBs) occurs in soils and aquatic sediments. In contrast to dechlorination of supplemented single congeners for which frequently ortho dechlorination has been observed, reductive dechlorination mainly attacks meta and/or para chlorines of PCB mixtures in contaminated sediments, although in a few instances ortho dechlorination of PCBs has been observed. Different microorganisms appear to be responsible for different dechlorination activities and the occurrence of various dehalogenation routes. No axenic cultures of an anaerobic microorganism have been obtained so far. Most probable number determinations indicate that the addition of PCB congeners, as potential electron acceptors, stimulates the growth of PCB-dechlorinating microorganisms. A few PCB-dechlorinating enrichment cultures have been obtained and partially characterized. Temperature, pH, availability of naturally occurring or of supplemented carbon sources, and the presence or absence of H(2) or other electron donors and competing electron acceptors influence the dechlorination rate, extent and route of PCB dechlorination. We conclude from the sum of the experimental data that these factors influence apparently the composition of the active microbial community and thus the routes, the rates and the extent of the dehalogenation. The observed effects are due to the specificity of the dehalogenating bacteria which become active as well as changing interactions between the dehalogenating and non-dehalogenating bacteria. Important interactions include the induced changes in the formation and utilization of H(2) by non-dechlorinating and dechlorinating bacteria, competition for substrates and other electron donors and acceptors, and changes in the formation of acidic fermentation products by heterotrophic and autotrophic acidogenic bacteria leading to changes in the pH of the sediments.     

Identification of the Proton Source for the Microbial Reductive Dechlorination of 2,3,4,5,6-Pentachlorobiphenyl

Microbially mediated reductive dechlorination of polychlorinated biphenyls (PCBs) in anaerobic sediments has been observed during laboratory experiments. Reductive dechlorination is a two-electron transfer reaction which involves the release of chlorine as a chloride ion and its replacement on the aromatic ring by hydrogen. The exact mechanism of the electron transfer for PCBs is unknown; however, this work shows that the source of the hydrogen atom is the proton (H⁺) from water.     

Theoretical investigation of the sequential reductive dechlorination pathways of chlorobenzenes and chloroanilines

The distribution of product isomers during the sequential reductive dechlorination of pentachloroaniline (PCA) and pentachlorobenzene (PeCB) was examined based on calculated thermodynamic, chromatographic, and electronic properties and then compared to the product distribution achieved by enrichment cultures. The dechlorination pathway analysis based on free energy considerations matched 78% and 67% of the experimental results for the sequential reductive dechlorination of chlorobenzenes (CBs) and chloroanilines (CAs), respectively. Chromatographic properties of CBs and CAs were able to explain some but not all of the reactions in the observed dechlorination pathways. Correlations between the observed dechlorination pattern and electronic properties of the parent compounds were able to explain most of the formation of the observed products. Experimentally observed sequential reductive dechlorination of CBs and CAs were similar to predicted dechlorination pathways based on the charge differential values calculated for the carbon–chloride bonds. Chlorine atoms were removed from the carbon atom that has the highest charge differential or the second highest charge differential. However, although thermodynamic, electronic as well as chromatographic properties of the CBs and CAs are certainly important factors, they may not be sufficient to completely describe the sequential microbial reductive dechlorination. Enzymatic specificity, as well as other factors (i.e., culture acclimation, environmental factors) should be considered for the interpretation of observed sequential reductive dehalogenation pathways of haloorganic compounds. This work provides the most comprehensive analysis to date of theoretical factors that control the sequential reductive chlorination of two homologous series of single‐ring chloroaromatic species. Biotechnol. Bioeng. 2010; 105: 574–587. © 2009 Wiley Periodicals, Inc.     

Dechlorination of Fenclor 54 and of a synthetic mixture of polychlorinated biphenyls by anaerobic microorganisms

Microorganisms obtained from a contaminated experimental soil were found to reductively dechlorinate the polychlorobiphenyls (PCBs) of ex-commercial Fenclor 54 and of a synthetic mixture of single congeners, under laboratory anaerobic conditions. The dechlorination rate and extent tended to increase as the chlorination degree of F 54 congeners increased. Several penta-chlorinated congeners temporarily accumulated during the final period of incubation. Dechlorination occurred primarily from the meta and para positions while ortho-sustituted congeners accumulated in the medium during incubation. The dechlorination pattern observed with these unacclimated microorganisms in both PCB mixtures could be only partially compared to patterns reported in the literature. The low product yield deriving from reductive dechlorination of PCBs, i.e. di-and tri-chlorinated biphenyls, and the slow rate of PCB biotransformation can be attributed to a lower dehalogenation capability of artificially contaminated soil microorganisms and, perhaps, also to the inadequacy of the adopted anaerobic medium. Correspondence to: F. Fava     

Redox and reduction potentials as parameters to predict the degradation pathway of chlorinated benzenes in anaerobic environments

Abstract The anaerobic degradation pathway of hexachlorobenzene starts with a series of reductive dehalogeneration steps. In the present paper it was evaluated whether the dehalogenation pathway observed in microbial ecosystems could be predicted by the redox potential and/or the reduction potential (the latter determined in dimethylsulfoxide) of the various potential intermediates. It was found that these two parameters suggest different pathways. The redox potential correctly predicts the dominant pathway observed in microbial systems, while the reduction potential does not. The redox potential of the various redox couples showed no correlation with the kinetic constants for the various dechlorination steps as determined with a quantitative structure-activity relationship developed for the environmental reductive dehalogenation of chlorinated aromatic compounds, even though both approaches predicted the same pathway.     

不同电子供体对2,4-二氯酚还原脱氯的影响

以葡萄糖、乙酸钠、Fe0、Fe0 葡萄糖、Fe0 乙酸钠作为电子供体,接种未驯化厌氧混合菌,考察2,4-二氯酚(2,4-DCP)的还原脱氯特性及Fe0作为电子供体的最佳作用条件与持续性特征.结果表明:与葡萄糖的作用相比,Fe0 葡萄糖可有效提高目标物脱氯效果;乙酸钠、Fe0及Fe0 乙酸钠均为有效电子供体,其中Fe0作为电子供体时目标物脱氯效果最佳,最佳作用条件为初始pH8.0,Fe0投加量2.0 g/L,4-CP为其主要脱氯中间产物;Fe0可持续供给2,4-DCP还原脱氯所需电子,而乙酸钠不断消耗后其脱氯效果与Fe0作为电子供体有明显差距.     

Bacterial dehalogenases: biochemistry, genetics, and biotechnological applications.

This review is a survey of bacterial dehalogenases that catalyze the cleavage of halogen substituents from haloaromatics, haloalkanes, haloalcohols, and haloalkanoic acids. Concerning the enzymatic cleavage of the carbon-halogen bond, seven mechanisms of dehalogenation are known, namely, reductive, oxygenolytic, hydrolytic, and thiolytic dehalogenation; intramolecular nucleophilic displacement; dehydrohalogenation; and hydration. Spontaneous dehalogenation reactions may occur as a result of chemical decomposition of unstable primary products of an unassociated enzyme reaction, and fortuitous dehalogenation can result from the action of broad-specificity enzymes converting halogenated analogs of their natural substrate. Reductive dehalogenation either is catalyzed by a specific dehalogenase or may be mediated by free or enzyme-bound transition metal cofactors (porphyrins, corrins). Desulfomonile tiedjei DCB-1 couples energy conservation to a reductive dechlorination reaction. The biochemistry and genetics of oxygenolytic and hydrolytic haloaromatic dehalogenases are discussed. Concerning the haloalkanes, oxygenases, glutathione S-transferases, halidohydrolases, and dehydrohalogenases are involved in the dehalogenation of different haloalkane compounds. The epoxide-forming halohydrin hydrogen halide lyases form a distinct class of dehalogenases. The dehalogenation of alpha-halosubstituted alkanoic acids is catalyzed by halidohydrolases, which, according to their substrate and inhibitor specificity and mode of product formation, are placed into distinct mechanistic groups. beta-Halosubstituted alkanoic acids are dehalogenated by halidohydrolases acting on the coenzyme A ester of the beta-haloalkanoic acid. Microbial systems offer a versatile potential for biotechnological applications. Because of their enantiomer selectivity, some dehalogenases are used as industrial biocatalysts for the synthesis of chiral compounds. The application of dehalogenases or bacterial strains in environmental protection technologies is discussed in detail.     

Comparison of anaerobic microbial communities from Estuarine sediments amended with halogenated compounds to enhance dechlorination of 1,2,3,4-tetrachlorodibenzo-p-dioxin

A suite of experiments were conducted to ascertain whether dehalogenation of a model dioxin compound could be stimulated in marine sediments by supplementation with halogenated analogues to enrich for dehalogenating bacteria and if growth by members of the Chloroflexi-like group was associated with dioxin removal. Five halogenated compounds (tetrachlorobenzene, tetrachloroanisole, tetrachlorophenol, tetrachlorobenzoic acid and trichloroacetophenone) were added with 1,2,3,4-tetrachlorodibenzo-p-dioxin (TeCDD) to estuarine sediments from four sites in San Diego Bay and the coast of southern New Jersey to test for dioxin dehalogenation. Most of the halogenated additives were found to stimulate dechlorination of the model dioxin. Molecular analysis of the bacterial population using 16S rRNA and reductive dehalogenase genes indicated that distinct microbial populations were enriched with each halogenated co-amendment. Additionally, Chloroflexi-like ribosomal genes associated with dehalogenation were detected. For example, quantitative real-time PCR analysis of 16S rRNA and reductive dehalogenase gene copy number in the microcosms showed a positive correlation with 1,2,3,4-TeCDD reductive dechlorination in coastal sediments amended with different halogenated additives. These results suggest that specific Chloroflexi-like microorganisms related to Dehalococcoides are involved in 1,2,3,4-TeCDD reductive dechlorination.     

Dehalogenation of lindane by a variety of porphyrins and corrins

The dehalogenation of lindane by a range of hemoproteins, porphyrins, and corrins has been tested under reducing conditions in the presence of dithiothreitol. In addition, a series of porphyrin-metal ion complexes have been prepared and have also been screened for the capacity to dehalogenate lindane. Hemoglobin, hemin, hematin, and chlorophyll alpha all catalyzed the dehalogenation of lindane, as did all of the corrins tested. The porphyrins which did not contain metal centers--coproporphyrin, hematoporphyrin, protoporphyrin, and uroporphyrin--were inactive. However, when these porphyrins were then complexed with Co, Fe, Mg, Mo, Ni, or V, lindane dehalogenation was observed. In all cases, the reaction proceeded by an initial dechlorination to produce tetrachlorocyclohexene, which was further dehalogenated to yield chlorobenzene as the end product.     

Dehalogenation of lindane by a variety of porphyrins and corrins.

The dehalogenation of lindane by a range of hemoproteins, porphyrins, and corrins has been tested under reducing conditions in the presence of dithiothreitol. In addition, a series of porphyrin-metal ion complexes have been prepared and have also been screened for the capacity to dehalogenate lindane. Hemoglobin, hemin, hematin, and chlorophyll alpha all catalyzed the dehalogenation of lindane, as did all of the corrins tested. The porphyrins which did not contain metal centers--coproporphyrin, hematoporphyrin, protoporphyrin, and uroporphyrin--were inactive. However, when these porphyrins were then complexed with Co, Fe, Mg, Mo, Ni, or V, lindane dehalogenation was observed. In all cases, the reaction proceeded by an initial dechlorination to produce tetrachlorocyclohexene, which was further dehalogenated to yield chlorobenzene as the end product.