红树林湿地多环芳烃的微生物降解

红树林(mangrove)是海陆交汇带重要的湿地生态系统,也是环境污染物蓄积与转化的热区.多环芳烃(polycyclic aromatic hydrocarbons,PAHs)因其环境蓄积特点在红树林生境中广泛分布,威胁生态系统健康,其降解转化是近年的研究重点.本文聚焦红树林湿地多环芳烃的微生物降解研究现状,从红树林生...     

芘在土壤中的共代谢降解研究

高分子量多环芳烃（PAHs)的降解通常以共代谢方式进行,研究比较了高分子量多环芳烃代表种类芘作为唯一C源和能源的降解过程和有共代谢底物存在下的降解过程,结果表明,25d后前者中芘的降解率57％,而后者中芘的降解率为80％,且有共代谢底物存在下,芘在降解过程中关衰期缩短;水扬酸,邻苯二甲酸,琥珀酸钠能作为共代谢底物提高芘的降解率,琥珀酸钠效果最好,芘和低要子量多环芳烃之间也有共代谢关系,菲促进了芘的降解,但萘未出现同样的结果,此外,这阐明了共代谢原理和适宜作高分子量多环芳烃共代谢底物的物质。     

多环芳烃(PAHs)微生物降解的原位表征方法

多环芳烃(polycyclic aromatic hydrocarbons,PAHs)是一类在环境中广泛存在的持久性有机污染物,微生物降解是去除环境中多环芳烃污染的主要途径。传统的有关PAHs微生物降解的研究主要依靠分离培养技术,难以准确认识PAHs微生物降解的原位过程及机制。近年来发展起来的原位表征方法可以在基因及单细胞水平研究PAHs在复杂环境中的微生物降解过程,能够原位表征具有PAHs降解功能的微生物及其功能基因和代谢活性,是阐明PAHs原位降解过程及分子机制的强有力的手段。该文综述了宏基因组技术(meta-genomics)、稳定同位素探针技术(stable isotope probe,SIP)、荧光原位杂交技术(fluorescence in situ hybridization,FISH)、拉曼光谱技术(Raman spectra)以及二次离子质谱技术(secondary ion mass spectrometry,SIMS)等原位表征技术在PAHs微生物降解研究领域的应用及其存在的问题和发展趋势等。PAHs微生物降解过程及机制的原位表征将为缓解与修复PAHs污染提供科学基础。     

多环芳烃污染土壤微生物修复研究进展

多环芳烃是我国土壤环境质量标准中要求严格管控的一类持久性有机污染物,利用微生物技术修复有机污染土壤具有绿色、经济等突出特点,应用前景广泛。目前多学科的协同发展和新技术的研究应用,为多环芳烃土壤微生物转化机制与污染生态过程等方面带来了新的认识,同时对修复技术的实际应用与调控提供了新的思考方向。本文以多环芳烃污染土壤微生物修复为主体,从污染土壤微生物修复应用技术、多环芳烃微生物降解特征、土壤体系污染物归趋规律与微生物作用及土壤污染微生物群落响应与研究技术等方面进行综合评述,并针对现存应用技术瓶颈和理论空白作进一步思考和展望。     

一株高效降解芘的细菌分离、鉴定及其降解效果

摘要：【目的】获得高效降解高分子量多环芳烃的细菌,并研究其对多环芳烃的降解能力。【方法】利用富集培养和芘升华平板方法,从焦化厂污染土壤中分离多环芳烃降解细菌,对分离菌株通过形态特征、16S rRNA基因和gyrb基因序列相似性分析进行鉴定,并研究该菌对高分子量多环芳烃（HMW-PAHs）的降解效果。【结果】筛选到一株能以芘、苯并蒽、屈、苯并芘、茚并芘、苯并苝、荧恩为碳源和能源生长并降解这些底物的菌株HBS1,该菌株的16S rRNA基因和gyrb基因序列与Gordonia amicalis的相应基因的相似     

多环芳烃污染土壤生物修复研究进展

多环芳烃 (Polycyclic aromatic hydrocarbons,PAHs) 是一类广泛分布于环境中的持久性污染物,结构稳定、难以降解,对生态环境和生物具有“三致”毒害性,其环境去除和修复备受关注。绿色、安全、经济的生物修复技术被广泛应用于PAHs污染土壤的修复。本文从土壤中PAHs的来源、迁移、归趋和污染水平总结了目前我国土壤多环芳烃污染的基本状况;归纳了具有PAHs降解作用的微生物、植物种类及机理;比较了微生物修复、植物修复和联合修复3类主要的生物修复技术。指出植物与微生物的互作机理的解析,抗逆菌株、植株的筛选与培育,实际应用的安全和效能评估应成为多环芳烃污染土壤修复领域未来的研究方向。     

石油污染土壤自然衰减过程多环芳烃降解基因动态特征

新疆石油污染土壤中微生物多环芳烃(polycyclic aromatic hydrocarbons,PAHs)降解功能基因研究甚少,且环境因子和功能基因之间相关性仍不清楚。【目的】揭示新疆石油污染砂质土壤自然衰减过程中多环芳烃降解关键基因结构和变化规律。【方法】以新疆准东油田为研究区,分析同一采油区不同石油污染年限土壤理化因子和多环芳烃含量变化,采用扩增子测序研究石油自然衰减过程中多环芳烃降解酶基因结构变化规律,利用Mental检验探讨其环境驱动因子。【结果】石油污染时间1年和3年的土壤中有多项理化指标与背景土存在显著性差异,而污染5年土壤与背景土之间仅2项指标具有显著性差异,随石油自然衰减逐渐恢复至正常。石油污染1年的土壤中16种多环芳烃除苊烯和?以外,其余14种多环芳烃均高于石油污染3年和5年土壤,多环芳烃总量和含油率污染1年土壤均显著高于污染3年和5年的土壤,多环芳烃会在污染后短时间内迅速被降解。扩增子测序结果显示,萘双加氧酶基因分类操作单元(operational taxonomic units,OTUs)序列随污染年限延长逐渐增多;芳环羟化双加氧酶基因OTUs序列BLAST(...     

微生物降解多环芳烃的研究进展

多环芳烃是一类长久存在于环境中,具有毒性、致突变与致癌等特性的环境优先污染物。本文对降解多环芳烃的微生物类群进行了阐述,介绍了在土壤与厌氧条件下细菌降解多环芳烃的研究情况,最后介绍了降解多环芳烃的相关酶类以及分子生物学的研究,并对消除环境中多环芳烃的相关生物技术提出展望。     

多环芳烃污染生物修复研究进展

多环芳烃(polycyclic aromatic hydrocarbon,PAHs)是一类对环境有严重危害的持久性有机污染物。具有高生物富集性、致癌性、致毒性和难降解性,修复治理PAHs污染环境备受国内外政府及学者的关注。目前主要采用物理、化学以及生物方法对多环芳烃污染的土壤和水体进行修复。其中生物修复是一种高效、经济和生态可承受的环保技术,具有成本低、无二次污染等优点。本文从植物修复、微生物修复以及植物-微生物联合修复方面,阐述了国内外生物修复PAHs污染的最新研究进展。指出了生物修复PAHs污染环境需要进一步解决的问题,并对未来发展趋势进行了展望。     

降解多环芳烃(PAHs)微生物研究进展

多环芳烃(PAHs)是一类有机污染物,来源广,对人体有害,微生物对环境中多环芳烃的降解有一定优越性。本文主要从降解微生物种类、影响因素、降解启动过程方面进行归纳和总结,针对以往研究工作的不足和存在的问题,指出了今后微生物降解多环芳烃研究中的重点,以求为研究者、管理者和决策者在应用时提供参考。     

Concurrent corticosteroid and phenanthrene transformation by filamentous fungus Cunninghamella elegans

A filamentous fungus Cunninghamella elegans IM 1785/21Gp which displays ability of 17alpha,21-dihydroxy-4-pregnene-3,20-dione (cortexolone) 11-hydroxylation (yielding epihydrocortisone (eF) and hydrocortisone (F)) and polycyclic aromatic hydrocarbons (PAHs) degradation, was used as a microbial eucaryotic model to study the relationships between mammalian steroid hydroxylation and PAHs metabolization. The obtained results showed faster transformation of phenanthrene in Sabouraud medium supplemented with steroid substrate (cortexolone). Simultaneously phenanthrene stimulated epihydrocortisone production from cortexolone. In phenanthrene presence the ratio between cortexolone hydroxylation products (hydrocortisone and epihydrocortisone) was changed from 1:5.1-6.2 to 1:7.6-8.4 in the culture without phenanthrene. Cytochrome P-450 content significantly increased after the culture supplementation by the second substrate, phenanthrene or cortexolone, adequately. To confirm the involvement of cytochrome P-450 in phenanthrene metabolism, the inhibition studies were performed. The cytochrome P-450 inhibitors SKF 525-A (1.5mM) and 2-methyl-1,2-di-3-pyridyl-1-propanone (metyrapone) (2mM) inhibited phenanthrene transformation by 80 and 62%, respectively. 1-aminobenzotriazole (1mM) completely blocked phenanthrene metabolism. The obtained results suggest a presence of connections between steroid hydroxylases and enzymes involved in PAH degradation in C. elegans.     

An insight into the origin and functional evolution of bacterial aromatic ring-hydroxylating oxygenases

Bacterial aromatic ring-hydroxylating oxygenases (RHOs) are multicomponent enzyme systems which have potential utility in bioremediation of aromatic compounds in the environment. To cope with the enormous diversity of aromatic compounds in the environment, this enzyme family has evolved remarkably exhibiting broad substrate specificity. RHOs are multicomponent enzymes comprising of a homo- or hetero-multimeric terminal oxygenase and one or more electron transport (ET) protein(s). The present study attempts in depicting the evolutionary scenarios that might have occurred during the evolution of RHOs, by analyzing a set of available sequences including those obtained from complete genomes. A modified classification scheme identifying four new RHO types has been suggested on the basis of their evolutionary and functional behaviours, in relation to structural configuration of substrates and preferred oxygenation site(s). The present scheme emphasizes on the fact that the phylogenetic affiliation of RHOs is distributed among four distinct 'Similarity classes', independent of the constituent ET components. Similar combination of RHO components that was previously considered to be equivalent and classified together [Kweon et al., BMC Biochemistry 9, 11 (2008)] were found here in distinct similarity classes indicating the role of substrate-binding terminal oxygenase in guiding the evolution of RHOs irrespective of the nature of constituent ET components. Finally, a model for evolution of the multicomponent RHO enzyme system has been proposed, beginning from genesis of the terminal oxygenase components followed by recruitment of constituent ET components, finally evolving into various 'extant' RHO types.     

Effects of carbon substrate enrichment and DOC concentration on biodegradation of PAHs in soil

AIMS: Two common reasons to explain slow environmental biodegradation of polycyclic aromatic hydrocarbons (PAHs), namely lack of appropriate carbon sources for microbial growth and limited bioavailability of PAHs, were tested in a laboratory bioassay using a creosote-contaminated soil. METHODS AND RESULTS: The soil, containing a total of 8 mg g-1 of 16 PAHs, was sieved and incubated in bottles for 45 days. The first explanation was tested by enrichment with the analogue anthracene and the non-analogue myristic acid, and both failed to stimulate degradation of all PAHs except anthracene. The second explanation was tested by addition of different concentrations of dissolved organic carbon (DOC), with effects depending on the DOC concentration and the molecular size of the PAH. The degradation was enhanced from 10 to 35% for 12 PAHs when the soil was saturated. The degraded amounts of individual PAHs were proportional to their concentration in the soil. CONCLUSIONS: The slow in situ degradation of PAHs was enhanced by more than three times by adding water as a solvent. Addition of DOC facilitated the degradation of four- to six-ring PAHs. SIGNIFICANCE AND IMPACT OF STUDY: Bioremediation of PAH-contaminated sites may be facilitated by creating water-saturated conditions but retarded by addition of other carbon substrates, such as analogue compounds.     

红树林厌氧环境对多环芳烃类有毒物的降解预测

红树林是连接陆地和海洋的重要生态系统,由于潮汐活动,氧化还原条件表现出明显的昼夜间的交替,这一生态体系中不但有大量的动植物种类,同时还有数量极高的不同种类的细菌,包括好氧和厌氧类型,厌养的硫酸(盐)还原菌已证实在降解多环芳烃有机物方面有其独特的生化优势,但从红树林中分离出的此类纯细菌还很少,在降解方面,已初步确定萘的厌氧降解途径异于好氧细菌,厌氧降解时的一系列代谢中间产物也有明显的专一性,羰基化反应是开始的一个重要步骤,而后的每步生化反应还有待进一步验证。从现有的结果可以看出,红树林中厌养的硫酸还原菌应在降解多环芳烃有机物中起到非常重要的作用。     

Degradation of high molecular weight polycyclic aromatic hydrocarbons by Pseudomonas cepacia

Summary When inoculated at high cell densities, three strains of Pseudomonas cepacia degraded the polycyclic aromatic hydrocarbons (PAHs) benzo[a]pyrene, dibenz[a,h]anthracene and coronene as sole carbon and energy sources. After 63 days incubation, there was a 20 to 30% decrease in the concentration of benzo[a]pyrene and dibenz[a,h]anthracene and a 65 to 70% decrease in coronene concentration. The three strains were also able to degrade all the PAHs simultaneously in a PAH substrate mixture containing three-, four-, five- and seven-benzene ring compounds. Furthermore, improved degradation of the five- and seven-ring PAHs was observed when low molecular weight PAHs were present.     

Of mothballs and old yellow enzymes

Polycyclic aromatic hydrocarbons (PAHs) are persistent and toxic environmental pollutants that accumulate in anoxic habitats. With the exception of naphthalene, nothing is known about the microbial degradation of PAH in these environments. The challenge that must be met in anaerobic PAH degradation is the destabilization of the resonance energy of the aromatic ring system, which requires electrons with very negative redox potentials. Estelmann et al. (2014) identify two enzymes from sulphate‐reducing bacteria that perform successive 2‐electron reductions of a coenzyme A thioester derivative of naphthalene. The first reduces 2‐naphthoyl‐CoA to 5,6‐dihydro‐2‐naphthoyl‐CoA and the second generates 5,6,7,8‐tetrahydro‐2‐naphthoyl‐CoA. Surprisingly, both enzymes are members of the ‘old yellow enzyme’ (OYE) family of flavoproteins. Neither uses adenosine triphosphate to achieve reduction of the aromatic ring. Typically, OYEs have flavin mononucleotide as cofactor and use nicotinamide adenine dinucleotide (phosphate) as reductant. Both ring reductases have flavin adenine dinucleotide and an iron‐sulphur cluster as additional cofactors. Evidence also suggests that in the sulphate‐reducing bacteria, these enzymes form a complex, allowing substrate channeling. The findings of this superb study represent unprecedented biochemistry. This work sheds light on how microbes meet the thermodynamic challenges of life at the redox limit.     

Anaerobic production and transformation of aromatic hydrocarbons and substituted phenols by ferulic acid-degrading BESA-inhibited methanogenic consortia

Abstract Sewage sludge-derived methanogenic enrichments degrading ferulic acid as sole carbon and energy source were partially inhibited with 2-bromoethanesulfonic acid. The various intermediates and products formed under inhibition of methanogenesis were studied using gas chromatography/mass spectrometry (GC/MS). In addition to aromatic, alicyclic, and aliphatic acids previously shown to be intermediates of ferulate degradation to CO₂ and CH₄, the following compounds were detected: toluene, ethylbenzene, phenol, p -cresol, 2-ethylphenol, catechol, and 3-hydroxy-4-ethylphenol. The character and the sequence of appearance of the compounds indicate that fermentative bacteria which initiate the anaerobic transformation of ferulic acid, in case of disruption of interspecies hydrogen transfer, dispose of electrons by converting part of the substrate to reduced derivatives. Aromatic hydrocarbons are further partially oxidized through hydroxylation of the ring (and, to a lesser extent, the side-chain), and partially reduced to saturated alicyclic rings. Some of these compounds seem to be gradually degraded to branched or straight- chain aliphatic acids. Some compounds, like catechol and ethylphenol, accumulate transiently or persistently in high concentrations (up to 16 mM carbon out of the initial concentration of 30 mM substrate carbon), indicating that hydroxylation of the aromatic ring might be an important metabolic reaction in these systems.     

Pleiotropic and Epistatic Behavior of a Ring-Hydroxylating Oxygenase System in the Polycyclic Aromatic Hydrocarbon Metabolic Network from Mycobacterium vanbaalenii PYR-1

Despite the considerable knowledge of bacterial high-molecular-weight (HMW) polycyclic aromatic hydrocarbon (PAH) metabolism, the key enzyme(s) and its pleiotropic and epistatic behavior(s) responsible for low-molecular-weight (LMW) PAHs in HMW PAH-metabolic networks remain poorly understood. In this study, a phenotype-based strategy, coupled with a spray plate method, selected a Mycobacterium vanbaalenii PYR-1 mutant (6G11) that degrades HMW PAHs but not LMW PAHs. Sequence analysis determined that the mutant was defective in pdoA2, encoding an aromatic ring-hydroxylating oxygenase (RHO). A series of metabolic comparisons using high-performance liquid chromatography (HPLC) analysis revealed that the mutant had a lower rate of degradation of fluorene, anthracene, and pyrene. Unlike the wild type, the mutant did not produce a color change in culture media containing fluorene, phenanthrene, and fluoranthene. An Escherichia coli expression experiment confirmed the ability of the Pdo system to oxidize biphenyl, the LMW PAHs naphthalene, phenanthrene, anthracene, and fluorene, and the HMW PAHs pyrene, fluoranthene, and benzo[a]pyrene, with the highest enzymatic activity directed toward three-ring PAHs. Structure analysis and PAH substrate docking simulations of the Pdo substrate-binding pocket rationalized the experimentally observed metabolic versatility on a molecular scale. Using information obtained in this study and from previous work, we constructed an RHO-centric functional map, allowing pleiotropic and epistatic enzymatic explanation of PAH metabolism. Taking the findings together, the Pdo system is an RHO system with the pleiotropic responsibility of LMW PAH-centric hydroxylation, and its epistatic functional contribution is also crucial for the metabolic quality and quantity of the PAH-MN.     

两种植物条件下土壤中矿物油和多环芳烃（PAHs）的生物修复研究

选择苜蓿草和水稻为供试植物,以污染物水平、有机以、专性细菌和真菌为调控因子,进行土壤中矿物油和PAHs的生物修复研究,结果表明,投肥对苜蓿草土壤中矿物油降解有促进作用,但对水稻土壤中矿物油降解无明显作用,投肥均使苜蓿草和水稻土壤中多环芒烃总量（11种列于美国EPA黑名单上的多环芳烃）降解率提高,这一降解促进效果在水稻土壤中好于苜蓿草土壤,有机肥量与苜蓿草根际土著真菌、细菌数量明显呈正相关,但仅与水稻根际土著细菌数量呈明显正相关,两种土壤中实测真菌和细菌总数均与试验投加专性真菌和细菌量无关,水稻土和苜蓿草土壤中3环多环芳烃的降解随投肥量增大而降解率提高,其在水稻土蓑中的效果好于苜蓿草土壤,投肥怪4环多环芳烃的降解并未产生有效作用。     

The catalytic pocket of the ring-hydroxylating dioxygenase from Sphingomonas CHY-1

Ring-hydroxylating dioxygenases are multicomponent bacterial enzymes that catalyze the first step in the oxidative degradation of aromatic hydrocarbons. The dioxygenase from Sphingomonas CHY-1 is unique in that it can oxidize a wide range of polycyclic aromatic hydrocarbons (PAHs). With a crystal structure similar to that of the seven other known dioxygenases, its catalytic domain features the largest hydrophobic substrate binding cavity characterized so far. Molecular modeling studies indicated that the catalytic cavity is large enough to accommodate a five-ring benzo[a]pyrene molecule. The predicted positions of this and other PAHs in the substrate binding pocket are consistent with the product regio- and stereo-selectivity of the enzyme.