A fungal phenoloxidase (tyrosinase) involved in pentachlorophenol degradation

Amylomyces rouxii eliminated 85% of initial pentachlorophenol (PCP) at 12.5 mg l^–1 when grown with 0.1 g tyrosine l^–1, but only 55% without tyrosine. Addition of tyrosine in the culture medium increased the monophenolase activity by 1.8-fold. Tyrosinase is thus indicated to be the phenoloxidase involved in PCP degradation by A. rouxii.     

Changes in soil microbial community structure and function following degradation in a temperate grassland

温带草原退化后土壤微生物群落结构和功能的变化 草原退化是草原生产力维持面临的一个重大挑战,这一过程显著影响着草原生态系统的能量流动和土壤养分变化过程,进而直接或间接地影响着土壤微生物。我们的研究目的首先是调查不同草原退化程度(即未退化、中度退化和严重退化)如何影响着内蒙古温带草原的土壤微生物组成、多样性和功能,其次是阐明哪些生物和非生物因素导致了这些变化。我们的研究主要通过高通量测序技术分析土壤微生物的群落组成,并且采用FAPROTAX工具和FUNGuild工具分别预测细菌群落和真菌群落的功能。研究发现：草原退化显著降低了土壤细菌的多样性,但对真菌多样性影响不大。地下生物量、土壤有机碳和总氮与细菌的多样性变化呈显著正相关关系。草原退化显著增加了绿弯菌门的相对丰度(由2.48%提高到8.40%),降低了厚壁菌门的相对丰度(由3.62%降低到1.08%)。其次,草原退化也显著增加了球囊菌门的相对丰度(从0.17%提高到1.53%),降低了担子菌门的相对丰度(从19.30%降低到4.83%)。致病菌的相对丰度在草原退化过程中显著下降。此外,草原退化对土壤细菌群落的功能有显著的影响,尤其是与土壤碳氮循环相关的土壤细菌群落。我们的结果表明,土壤细菌群落对草原退化的响应比真菌群落更敏感。     

环境雌激素的微生物降解

雌激素作为环境内分泌干扰物的一类重要物质,生物降解法是最为经济、最为绿色和最为适用的去除方法。通过分析雌激素的主要来源和危害、国内外雌激素降解菌的筛选与鉴定、类固醇雌激素降解酶的表达与检测、雌激素降解菌的基因组学特征以及雌激素的降解途径和机制,进一步阐述了雌激素生物降解的研究现状与进展,对未来的研究工作作出了展望。     

五氯酚在土壤生态系统中的环境行为研究进展

五氯酚在世界范围内广泛使用,在曾经使用过五氯酚的国家和地区的土壤和沉积物可以检测到五氯酚的残留,甚至残留五氯酚可通过食物链危及人类健康。综述了五氯酚在土壤生态系统中的吸附机制与影响因素、化学降解、生物降解及其影响因素、生物富集、生态毒理机理和效应。同时结合目前国内外的研究现状,提出了五氯酚在土壤生态系统中环境行为新的研究方向。     

Biodegradation of pentachlorophenol by white rot fungi under ligninolytic and nonligninolytic conditions

The roles of lignin peroxidase, manganese peroxidase, and laccase were investigated in the biodegradation of pentachlorophenol (PCP) by several white rot fungi. The disappearance of pentachlorophenol from cultures of wild type strains,P. chrysosporium, Trametes sp. andPleurotus sp., was observed. The activities of manganese peroxidase and laccase were detected inTiametes sp. andPleurotus sp. cultures. However, the activities of ligninolytic enzymes were not detected inP. chrysosporium cultures. Therefore, our results showed that PCP was degraded under ligninolytic as well as nonligninolytic conditions. Indicating that lignin peroxidase, manganese peroxidase, and laccase are not essential in the biodegradation of PCP by white rot fungi.     

阿维菌素在土壤中的微生物降解研究

运用恒温培养法研究了阿维菌素在土壤中的降解动力学．结果表明,非生物+微生物降解、非生物降解及微生物降解的半衰期分别为34．8、277．3和49．9d,说明阿维菌素在土壤中的降解主要由微生物引起．从试验土壤中分离到1株高效降解阿维菌素的菌株,经16S rDNA鉴定为嗜麦芽寡养单胞菌(Stenotrophomonas maltrophilia)．从该降解菌中提取的粗酶液米氏常数(Km)为6．78nmol·ml^-1,最大降解速率为81．5nmol·min^-1·mg^-1。     

Degradation of p-nitrophenol and pentachlorophenol mixtures by Sphingomonas sp. UG30 in soil perfusion bioreactors

The degradation of mixtures of pentachlorophenol (PCP) and p-nitrophenol (PNP) were evaluated in pure cultures of Sphingomonas sp. UG30, statically incubated soils (60% water-holding capacity) and soil perfusion bioreactors where encapsulated cells of UG30 were used as a soil inoculant. In pure-culture studies, conditions were optimized for mineralization of PCP and PNP mixtures at concentrations of 30 mg l⁻¹ each. Optimum in vitro mineralization of PCP and PNP mixtures by UG30 was facilitated using ammonium phosphate as a nitrogen source, while inhibition was observed with ammonium nitrate. The bioreactor system used columns containing soil treated with mixtures of 100, 225 or 500 mg kg⁻¹ of PCP and PNP. Rapid dissipation of both substrates was observed at the 100 mg kg⁻¹ level. Inoculation with UG30 enhanced PCP degradation at the 100 mg kg⁻¹ level in bioreactors but not in static soil microcosms. At higher PCP and PNP concentrations (225 mg kg⁻¹), occasional complete degradation of PNP was observed, and PCP degradation was about 80% compared to about 25% in statically incubated soils after 20 days at 22°C. There was no additional degradation of the PCP and PNP mixtures attributable to inoculation with encapsulated cells of UG30 in either soil system at concentrations of 225 or 500 mg kg⁻¹. Journal of Industrial Microbiology & Biotechnology (2000) 25, 93–99. Received 25 February 2000/ Accepted in revised form 07 June 2000     

石油基塑料的微生物降解

石油基塑料种类繁多、数量巨大、应用广泛,常见的有聚乙烯(PE)、聚丙烯(PP)、聚苯乙烯(PS)、聚氯乙烯(PVC)、聚对苯二甲酸乙二醇酯(PET)、聚氨酯(PUR)等。这些合成塑料因其高分子量、高疏水性及高化学键能的特点难以被微生物降解,从而在环境中长期存在和累积,"白色污染"已经成为一个全球性问题。因此安全经济的微生物降解合成塑料是人类面临的一个选择和难题。文中从微生物资源及相关酶学研究方面综述了聚苯乙烯、聚乙烯、聚丙烯、聚氨酯、聚对苯二甲酸乙二醇酯和聚氯乙烯这6种石油基塑料的生物降解的研究现状。目前关于上述6种石油基塑料的微生物降解研究依然大多停留在微生物资源的寻找中,已发现的具备相关能力的菌株种类较少,并且微生物降解效率均非常缓慢;对于其降解机理及关键基因和酶的研究比较少。文中为进一步开展塑料生物降解研究,寻找高效的塑料降解菌株资源以及进一步在遗传、分子和生化水平研究塑料生物降解机理研究,从而最终实现合成塑料的彻底降解和高值化利用提供了借鉴。     

木质纤维素的微生物降解

木质纤维素广泛存在于自然界中,因结构复杂,其高效降解需要多种微生物的协同互作,由于参与木质纤维素降解的微生物种类繁多,其协同降解机理尚不完全明确。随着微生物分子生物学和组学技术的快速发展,将为微生物协同降解木质纤维素机制的研究提供新的方法和思路。笔者前期研究发现,细菌复合菌系在50℃下表现出强大的木质纤维素降解能力,菌系由可分离培养和暂时不可分离培养细菌组成,但是可分离培养细菌没有降解能力。通过宏基因组和宏转录组研究表明,与木质纤维素降解相关的某些基因表达量发生显著变化,通过组学方法有可能更加深入解释微生物协同降解木质纤维素的微生物学和酶学机理。文中从酶、纯培养菌株和复合菌群三个方面综述了木质纤维素微生物降解研究进展,着重介绍了组学技术在解析复合菌群作用机理方面的现状和应用前景,以期为探索微生物群落协同降解木质纤维素的机理提供借鉴。     

环境内分泌干扰物雌激素的微生物降解研究进展

雌激素是一类重要的环境内分泌干扰物。微生物降解是一种去除环境雌激素与进行环境修复的最绿色、环保、经济的方法。本文从分析雌激素的主要来源和危害、归纳国内外已报道的雌激素降解菌、总结雌激素降解的相关基因和组学研究进展、阐述雌激素的降解通路和降解机制这4个方面,概括阐述了环境雌激素的微生物降解作用,并对未来雌激素降解研究的主要内容与方向提出展望。     

聚氨酯塑料的微生物降解

未被合理处置的废塑料污染已成为全球性的环境问题,探索塑料废弃物的无害化处理技术势在必行。近来,研究证实了自然界中存在可以降解塑料的微生物及酶。利用微生物或酶对废塑料进行生物处理成为可能。聚氨酯塑料(Polyurethane,PUR)是广泛应用的通用塑料之一,其废弃物量已占到所有废塑料总体积的30%。文中将PUR塑料发明应用70年来有关微生物降解的研究进行了全面综述,对PUR塑料降解真菌、细菌、降解基因与酶、降解产物及相关的生物处理技术系统等进行了总结与分析,并对实现PUR废塑料高效生物处理需解决的关键科学问题进行了展望。     

Microbial DNA profiling by multiplex terminal restriction fragment length polymorphism for forensic comparison of soil and the influence of sample condition

Aims:  To evaluate: (i) the impact of air-drying on bacterial, archaeal and fungal soil DNA profiles and (ii) the potential use of multiplex-terminal restriction fragment length polymorphism (M-TRFLP) as a tool for forensic comparison of soil.
Methods and Results:  An M-TRFLP approach was used to profile bacterial, archaeal and fungal DNA profiles from five different soil sites. Air-drying soil significantly reduced the quantity of DNA but the number of operational taxanomic units (OTU) was unaffected. The impact of air-drying on soil DNA profiles was dependent on soil site and microbial primers. Fungal profiles were altered the least by air-drying. For prokaryotic profiles, air-drying altered the relative similarity/dissimilarity between soil sites. The M-TRFLP approach was more discriminatory compared with soil colour and single-taxa profiling, but did not significantly improve resolution between two similar soils.
Conclusions:  Of those tested, soil fungi were potentially the more robust target for application to soil forensic studies as they were altered less by air-drying and provided clear discrimination of soils from different sites. The M-TRFLP method demonstrated potential to achieve greater resolution, discriminating the soil sites based on both bacterial and fungal components.
Significance and Impact of the Study:  Soil DNA profiling has potential as a forensic tool, but sample condition and the appropriate selection of microbial target taxa must be considered.     

聚乙烯塑料的微生物降解

聚乙烯(polyethylene,PE)是产量最大的通用塑料之一,通常被加工成一次性包装材料(包括塑料袋及容器)和农用薄膜等。PE塑料的广泛应用导致大量PE废弃物的累积,对生态环境造成严重的威胁。自20世纪70年代以来,一些研究陆续报道了PE塑料被微生物降解的现象,并从土壤、海洋、垃圾堆置点及昆虫肠道等生境中分离筛选到了若干种具有一定PE塑料降解能力的菌株,而且发现一些单加氧酶、过氧化物酶和漆酶等氧化还原酶对PE塑料具有氧化降解能力。这些研究为发展PE塑料废弃物生物降解处理技术提供了一定的依据。本文总结和分析了PE塑料降解微生物的分离和筛选方法,以及已报道的PE塑料降解微生物和降解酶的研究进展,以期为进一步研究PE塑料的微生物降解机理和处理技术提供参考。     

Degradation of pentachlorophenol by the white rot fungus Phanerochaete chrysosporium grown in ammonium lignosulphonate media

Removal and degradation of pentachlorophenol (PCP) by Phanerochaete chrysosporium in static flask cultures was studied using ammonium lignosulphonates (LS), a waste product of the papermill industry, as a carbon and nitrogen source. After 3 days, cultures of P. chrysosporium grown in either a 2% LS (nitrogen-sufficient) medium or a 0.23% LS and 2% glucose (nitrogen-deficient) medium removed 72 to 75% of PCP, slightly less than the 95% removal seen using nitrogen-deficient glucose and ammonia medium. PCP dehalogenation occurred despite the fact that extracellular enzyme (LiP) activity, measured by a veratryl alcohol oxidation assay and by PCP disappearance in cell-free extracts, was inhibited by LS. This inactivation of LiP likely contributed to the lower percent of PCP dehalogenation observed using the LS media. In order to better understand the relationship between PCP disappearance and dehalogenation, we measured the fate of the chlorine in PCP. After 13 days, only 1.8% of the initial PCP added was recoverable as PCP. The remainder of the PCP was either mineralized or transformed to breakdown intermediates collectively identified as organic halides. The largest fraction of the original chlorine (58%) was recovered as organic (non-PCP) halide, most of which (73%) was associated with the cell mass. Of the remaining chlorine, 40% was released as chloride ion, indicating a level of dehalogenation in agreement with previously reported values.     

Microbial degradation of chlorinated benzenes

Chlorinated benzenes are important industrial intermediates and solvents. Their widespread use has resulted in broad distribution of these compounds in the environment. Chlorobenzenes (CBs) are subject to both aerobic and anaerobic metabolism. Under aerobic conditions, CBs with four or less chlorine groups are susceptible to oxidation by aerobic bacteria, including bacteria (Burkholderia, Pseudomonas, etc.) that grow on such compounds as the sole source of carbon and energy. Sound evidence for the mineralization of CBs has been provided based on stoichiometric release of chloride or mineralization of (14)C-labeled CBs to (14)CO(2). The degradative attack of CBs by these strains is initiated with dioxygenases eventually yielding chlorocatechols as intermediates in a pathway leading to CO(2) and chloride. Higher CBs are readily reductively dehalogenated to lower chlorinated benzenes in anaerobic environments. Halorespiring bacteria from the genus Dehalococcoides are implicated in this conversion. Lower chlorinated benzenes are less readily converted, and mono-chlorinated benzene is recalcitrant to biotransformation under anaerobic conditions.     

Phylogeny of Sphingomonas species that degrade pentachlorophenol

Four pentachlorophenol (PCP)-degrading bacteria isolated from geographically diverse areas have been examined in detail as regards their physiology and phylogeny. According to traditional biochemical methods, these strains had been classified as members of the genera Arthrobacter, Flavobacterium, Pseudomonas, and Sphingomonas. The PCP degradation pathway has been studied extensively in Sphingomonas (Flavobacterium) sp strain ATCC 39723 and the first three degradation steps catalyzed by a PCP-4-monooxygenase (PcpB) and a reductive dehalogenase (PcpC) that functions twice are well established. A fourth step appears to involve ring-fission of the aromatic nucleus (PcpA). Molecular analyses revealed that the PCP degradation pathway in these four strains was rather conserved, leading to a phylogenetic analysis using 16S rDNA. The results revealed a much closer phylogenetic relationship between these organisms than traditional classification indicated, placing them into the more recently established genus Sphingomonas where they may even represent a single species. With 16S rDNA analysis, many bacterial isolates involved in degradation of xenobiotic compounds that were previously classified into diverse genera have been reclassified into the genus Sphingomonas. Received 14 April 1999/ Accepted in revised form 20 July 1999     

Toxicity of pentachlorophenol for micromycetes isolated from freshwater sediments

In order to evaluate the impact of xenobiotics on fungal communities of sediments, lentic ecosystems have been reconstituted in the laboratory and examined before as well as after contamination.Microcosms (2 litres) have been treated with three different concentrations of pentachlorophenol (PCP) (3, 100, and 1000 mg l^-1) at 22 degC. The fungal strains present in the sediments were first isolated and identified before contamination with PCP. Most of the strains belonged to Deuteromycetes (mainly Mucedinaceæ and Dematiaceæ) with a predominance of the genus Penicillium, and Ascomycetes were found in exceptional abundance. New isolations and identifications were made on days 1, 2, 4, 8, and 15 after contamination with PCP. The sensitivity of the various species were determined as a function of the level of PCP and the duration of contamination. Ascomycetes was particularly resistant to high concentrations of PCP. The toxicity of the xenobiotic towards strains belonging to various taxonomic groups is discussed, as well as the possibility that some of them may be useful for biodegradation and/or bioremediation processes, or as bioindicators of polluted sites.Groupe pour l'Étude du Devenir des Xénobiotiques dans l'Environnement (GEDEXE)Laboratoire de Botanique, Cryptogamie, Biologie Cellulaire et Génétique.     

微生物菌剂对秸秆降解及秸秆中微生物变化规律的影响

为明确不同微生物菌剂对秸秆降解率及秸秆降解过程中秸秆周围微生物变化规律的影响，在温室大棚内进行不同微生物菌剂降解秸秆试验。以玉米秸秆为基质，设玉米秸秆（对照组）、玉米秸秆+哈茨木霉（Trichoderma harzianum）（处理1）、玉米秸秆+哈茨木霉+地衣芽胞杆菌(Bacillus licheniformis)（处理2）三个处理。研究结果显示，单独用哈茨木霉处理可显著提高秸秆降解率，前期作用尤其明显；在秸秆降解的前30 d，秸秆降解率与秸秆中可培养真菌、细菌、放线菌呈显著正相关。不同微生物菌剂对秸秆中可培养微生物数量变化有显著影响，单独接种哈茨木霉后，秸秆中可培养真菌、细菌、放线菌数在前30 d显著高于对照组和混合菌剂处理，不同处理可培养活菌数均在90～120 d达到峰值，然后开始下降。研究结果表明，不同微生物菌剂对秸秆降解有显著影响，单独用哈茨木霉处理可显著提高秸秆降解率；不同处理对秸秆中可培养真菌、细菌、放线菌的影响有显著差异。可为秸秆降解与微生物相关性研究提供参考。     

Microbial communities involved in anaerobic degradation of alkanes

Saturated hydrocarbons are quantitatively the most abundant fraction among all petroleum hydrocarbons. Significant advances have been made in the understanding of the anaerobic biodegradability of alkanes in terms of the microorganisms involved and the biochemical pathways over the past two decades. They can be used as carbon and energy sources by diverse physiological groups of microorganisms (isolates or consortia) grown under chlorate-reducing, nitrate-reducing, sufidogenic or methanogenic conditions. Two general biochemical mechanisms have been proposed for the initial activation of alkanes including addition of fumarate and carboxylation. However, glycyl radical enzymes dependent fumarate addition which yields alkyl-substituted succinate appear to be the most commonly shared mechanism for the anaerobic attack of alkanes under various redox conditions by phylogenetically diverse microorganisms. The genes encoding the candidate alkylsuccinate synthase have been recently described in alkane-degrading sulfate- and nitrate-reducers as well as in hydrocarbon-rich environments. Alternative mechanisms may also be available depending on the alkane-degrading microbial community and electron acceptors utilized.