氯氰菊酯高效降解菌GF31的生长特性及其对污染土壤的修复能力

利用高效降解菌对受污土壤进行生物强化修复是提高修复效果的可行途径之一,其中降解菌的竞争力与适应性是决定强化修复过程成败的关键因素.以实验室保藏的1株氯氰菊酯高效降解菌--铜绿假单胞菌CF31为对象,开展菌株生长特性及其对污染土壤修复的实验研究.结果表明,菌株GF31生物活性高,环境适应力强,在温度15～35℃,pH值5...     

高效氯氰菊酯降解菌群的结构分析及其降解特性研究

以连作10年以上的棉田土壤为材料,以高效氯氰菊酯为唯一碳源,驯化获得能稳定传代并持续降解高效氯氰菊酯的LZ1菌群,对菌群中可培养的细菌进行分离鉴定,最后对其降解高效氯氰菊酯的特性进行分析。结果表明,LZ1菌群中可分离、纯化优势菌株12株,经16SrRNA序列分析,其中10株与铜绿假单胞菌(Pseudomonas aeruginosa)相似性达99%,1株与无色杆菌(Achromobacter mucicolens)相似性达99%。高效氯氰菊酯最佳反应条件为高效氯氰菊酯初始质量浓度250mg/L、温度27℃、pH7.0、装样量200mL。在最佳反应条件下培养的LZ1菌群24h时对高效氯氰菊酯的降解率可达68.81%,132h时的降解率可达92.39%,且LZ1菌群对高效氯氰菊酯的4种异构体没有明显的降解特异性。     

氯氰菊酯降解菌CY22-7的分离鉴定及降解特性研究

从采集的多种土壤样品中.获得了19株在3种筛选平板上均生长良好的候选氯氰菊酯降解菌.通过生理生化特性鉴定和16S rDNA序列的测定及比对.将其中的氯氰菊酯降解菌CY22-7鉴定为中华根瘤菌属(Sinorhizobium sp.).同时,对氯氰菊酯降解菌CY22-7的降解特性研究结果表明:(1)氯氰菊酯降解菌CY22-7以5%(体积分数)的接种鼍接种到氯氰菊酯起始质量浓度为100 mg/L的氯氰菊酯乙醇培养基后,于200 r/min、30℃摇床中培养.经过6 d的培养,氯氰菊酯降解菌CY22-7降解了约60%的氯氰菊酯.(2)加人外源营养物质有利于促进氯氰菊酯的降解.其中,葡萄糖和酵母提取物的促进作用最为明显.(3)氯氰菊酯降解菌CY22-7降解氯氰菊酯的最适温度为30℃,最佳pH为6.0.     

氯氰菊酯的酶促降解研究

从氯氰菊酯高效降解真菌镰孢霉属（Fusarium）菌株TS-203中提取了降解酶，研究了降解酶对氯氰菊酯的降解特性。结果表明，胞内酶对氯氰菊酯的降解率高达59．8％，细胞碎片对氯氰菊酯的降解率为47．6％，而由（NH。）：sO。沉淀法提取到的胞外酶对氯氰菊酯的降解率仅为10．3％，由此确定菌株TS-203产生的降解酶为胞内酶。以牛血清白蛋白为标准蛋白测得胞内粗酶液中可溶性蛋白质含量为3．24mg／mL；该酶对氯氰菊酯酶促降解的最适pH为7．0，最适温度为30℃，降解酶的米氏常数K。为6．8120×10^-4mmol／mL，最大反应速度Vmax为1．1799×10^-4mmol／min。研究结果表明，该酶具有较好的热稳定性和pH稳定性，对热和pH均具有较好的耐受力，对氯氰菊酯降解效果较好。     

Rhodobacter sp. NP25b菌株缺氧降解壬基酚聚氧乙烯醚的研究

从城市污水处理厂活性污泥中分离得到一株能够在缺氧条件下以壬基酚聚氧乙烯醚(NPEOs)为惟一碳源和能源生长的菌株NP25b.经生理生化鉴定和16S rRNA基因序列分析,该菌株属于红细菌属(Rhodobacter sp.),对该菌株降解NPEOs的特性进行了研究.结果表明,在缺氧条件下,菌株NP25b在7 d内对初始底物浓度为400 mg/L NPEOs的降解率可达84%.利用液相色谱-质谱(LC-MS)和气相色谱-质谱(GC-MS)对NPEOs降解中间产物进行了分析,结果表明,主要降解产物为短链NPEOs和壬基酚聚氧乙烯醚乙酸(NPECs),其中包括具有较强内分泌干扰效应的NP1EO.该菌株能够代谢含有疏水基团的聚氧乙烯醚类表面活性剂,例如辛基酚聚氧乙烯醚和脂肪醇聚氧乙烯醚.推测菌株NP25b降解NPEOs是通过乙氧基(EO)链末端氧化后逐步切割完成的.     

石油降解菌Pseudomonas stutzeri TH-31的分离与降解条件优化

使用稀释富集法,从大港油田采油废水处理站生化池中定向快速的分离出多株高效石油降解菌株。对分离获得的1株优势菌株进行生理生化和分子生物学鉴定,显示属于Pseudomonas stutzeri的1个新菌株,命名为Pseudomonas stutzeri TH-31。通过批次实验,对菌株TH-31的生长条件和石油降解条件进行了优化,经过条件优化,P.stutzeri TH-31在初始p H 7,原油投加量为300 mg/L,35℃培养5 d后,获得最高石油烃降解率92.7%。     

嗜热栖热菌降解氟喹诺酮类抗生素

氟喹诺酮类抗生素在各种环境基质中积累造成的生态和耐药基因污染等问题已引起广泛的关注。为了能够有效去除环境中氟喹诺酮类抗生素污染并且探究其生物代谢途径,利用嗜热菌Thermus sp. C419在高温(70℃)条件下降解2种典型的氟喹诺酮类抗生素(诺氟沙星和恩诺沙星),分析了菌株C419对这2种药物在单一和混合添加时的降解特性;通过UPLC-MS/MS检测了其相关的降解产物,并推测了可能的代谢途径;利用平板扩散法对生物降解后的氟喹诺酮类药物进行抑菌活性测定。结果表明:氟喹诺酮类化合物可被菌株C419有效降解,降解率为60%～80%;该生物降解过程符合一级动力学模型,培养基中氟喹诺酮类化合物浓度越高,降解率越高,降解半衰期越短;菌株C419对诺氟沙星的生物降解有3条可能的降解途径和7种降解产物,对恩诺沙星的生物降解有4条可能的降解途径和6种降解产物。此外,与2种药物的母体化合物相比,生物降解后药物对不同细菌的抗菌活性均有一定程度的降低,这说明嗜热菌株C419在热环境中去除氟喹诺酮类污染物方面可能会具有良好的实用性和应用前景。     

染料结晶紫降解菌分离及其特性研究

从印染废水中筛选分离得到结晶紫(CV)降解菌株H,并对其脱色条件和机理进行了研究.结果表明,在外加生长基质条件下,菌株H不仅对CV降解脱色而且能降解中间产物.在pH 7.0、35℃、摇床转速180 r/min条件下,菌株H有最佳的脱色效果.对于20 mg/L CV溶液,生长细胞降解0.5 h,脱色率可达95%.休眠细胞对CV的降解规律也进行了对比探讨.菌株H对CV脱色过程遵循一级动力学方程,脱色速率常数k随CV浓度升高而降低,生长细胞的k值约为休眠细胞的6～7倍.     

白腐菌P.sordida YK-624及其锰过氧化物酶对2-氯酚的转化

利用白腐菌P.sordida YK-624和产自该菌株的锰过氧化物酶(MnP)处理2-氯酚(2-CP).结果表明,当2-CP质量浓度为50mg/L时,培养5d, P.sordida YK-624能降解50.42%的2-CP;采用300 U的MnP处理150mg/L的2-CP, 24h时2-CP降解率可达75.00%.利用紫外-可见分光光度计、红外光谱仪和高效液相色谱仪对MnP处理过程中的产物进行了分析,表明MnP氧化2-CP过程中有多种产物生成,可能的产物包括醌类和聚合产物.     

氯氰菊酯降解真菌的筛选及其降解特性研究

用富集培养的方法从农药厂污水排放口的污泥中分离筛选到3株以氯氰菊酯为唯一碳源进行生长的真菌,分别命名为TS-203、TS-205和TS-306。经鉴定3株真菌分别属于土生曲霉组(Aspergillus terreus)、毛链孢属(M onilochaetes)和镰孢霉属(Fusarium)。分别测定了不同碳源、pH、培养温度及氯氰菊酯浓度对真菌生长量和降解能力的影响。结果表明,以氯氰菊酯为唯一碳源且浓度为50~150 mg/L,pH 6.0~8.0,培养温度30~40℃时,真菌的生长量较大,降解效果较好。本研究对氯氰菊酯农药残留的生物修复研究和应用具有十分重要的意义,可为今后治理氯氰菊酯残留污染提供参考。     

Naphthalene metabolism in Nocardia otitidiscaviarum strain TSH1, a moderately thermophilic microorganism

The thermophilic bacterium Nocardia otitidiscaviarum strain TSH1, originally isolated in our laboratory from a petroindustrial wastewater contaminated soil in Iran, grows at 50 degrees C on a broad range of hydrocarbons. Transformation of naphthalene by strain TSH1 which is able to use this two ring-polycyclic aromatic hydrocarbon (PAH) as a sole source of carbon and energy was investigated. The metabolic pathway was elucidated by identifying metabolites, biotransformation studies and monitoring enzyme activities in cell-free extracts. The identification of metabolites suggests that strain TSH1 initiates its attack on naphthalene by dioxygenation at its C-1 and C-2 positions to give 1,2-dihydro-1,2-dihydroxynaphthalene. The intermediate 2-hydroxycinnamic acid, characteristic of the meta-cleavage of the resulting diol was identified in the acidic extract. Apart from typical metabolites of naphthalene degradation known from mesophiles, benzoic acid was identified as an intermediate for the naphthalene pathway of this Nocardia strain. Neither phthalic acid nor salicylic acid metabolites were detected in culture extracts. Enzymatic experiments with cell extract showed the catechol 1,2-dioxygenase activity while transformation of phthalic acid and protocatechuic acid was not observed. The results of enzyme activity assays and identification of benzoic acid in culture extract provide strong indications that further degradation goes through benzoate and beta-ketoadipate pathway. Our results indicate that naphthalene degradation by thermophilic N. otitidiscaviarum strain TSH1 differs from the known pathways found for the thermophilic Bacillus thermoleovorans Hamburg 2 and mesophilic bacteria.     

Complete degradation of butyl benzyl phthalate by a defined bacterial consortium: role of individual isolates in the assimilation pathway

Two bacterial strains, in consortium, were isolated by enrichment techniques from municipal waste-contaminated soil, which utilized butyl benzyl phthalate (BBP) as the sole carbon source. One of the isolates was identified as Arthrobacter sp. strain WY and the other one as Acinetobacter sp. strain FW based on the morphological, nutritional and biochemical characteristics and 16S rRNA sequence analysis. Various metabolites of BBP engendered by Arthrobacter sp. strain WY were isolated and identified by a combination of chromatographic and spectrophotometric analyses, which revealed a pathway involving monobutylphthalate (MBuP), monobenzyl phthalate (MBzP), phthalic acid and protocatechuic acid. The protocatechuic acid in turn was processed by ortho-cleavage dioxygenase to form beta-carboxy-cis,cis-muconate, ultimately leading to the TCA cycle. The Arthrobacter sp. strain WY could not utilize the hydrolyzed alcohols of BBP. On the other hand, the Acinetobacter sp. strain FW, which by itself could not utilize BBP as the sole carbon source, is capable of utilizing the hydrolyzed alcohols of BBP. Benzyl alcohol was found to be metabolized by the Acinetobacter sp. strain FW via benzaldehyde, benzoic acid and catechol. Catechol was further degraded by ortho-cleavage dioxygenase to cis,cis-muconic acid and subsequently to muconolactone leading to beta-ketoadipate pathway. Moreover, the Acinetobacter sp. strain FW metabolized 1-butanol through butyraldehyde and butyric acid leading to the tricarboxylic acid cycle via beta-oxidation pathway. This is the first report on the complete degradation of BBP by a defined consortium describing the role of its individual constituents in the BBP assimilation pathway.     

Biodegradation of beta-cypermethrin by a novel Azoarcus indigens strain HZ5

A novel strain HZ5 was isolated from the activated sludge of a pesticide manufacturer in Hangzhou, which was capable of degrading beta-cypermethrin (beta-CP). Based on its physiological characteristics and analysis of 16S rDNA gene, strain HZ5 was identified as Azoarcus indigens, which was a new genus that can degrade beta-CP effectively. Strain HZ5 could degrade beta-CP over a wide range of temperature (20 to 40°C) and pH (5.5 to 9.0), and the optimal temperature and pH were 30°C and 7.0. The highest degradation rate was approximately 70% of 50 mg/L beta-CP within 144 h at pH 7.0 and 30°C in MSM. An additional carbon source could enhance the biodegradation of beta-CP. Studies on biodegradation of the beta-CP showed no significant enantioselectivity. During the process, two main intermediate metabolites were produced by strain HZ5 and determined as 3-phenoxybenzaldehyde and 3-phenoxybenzoic acid by gas chromatography-mass spectrometry (GC-MS) analysis. The results indicated that strain HZ5 may have potential application in bioremediation of beta-CP polluted environment.     

Biodegradation of malathion, α- and β-endosulfan by bacterial strains isolated from agricultural soil in Veracruz,Mexico

The objective of this study was to evaluate the capacity of two bacterial strains isolated, cultivated, and purified from agricultural soils of Veracruz, Mexico, for biodegradation and mineralisation of malathion (diethyl 2-(dimethoxyphosphorothioyl) succinate) and α- and β-endosulfan (6,7,8,9,10,10-hexachloro-1,5,5a,6,9,9a-hexahydro-6-9-methano-2,4,3-benzodioxathiepine-3-oxide). The isolated bacterial strains were identified using biochemical and morphological characterization and the analysis of their 16S rDNA gene, as Enterobacter cloacae strain PMM16 (E1) and E. amnigenus strain XGL214 (M1). The E1 strain was able to degrade endosulfan, whereas the M1 strain was capable of degrading both pesticides. The E1 strain degraded 71.32% of α-endosulfan and 100% of β-endosulfan within 24 days. The absence of metabolites, such as endosulfan sulfate, endosulfan lactone, or endosulfan diol, would suggest degradation of endosulfan isomers through non-oxidative pathways. Malathion was completely eliminated by the M1 strain. The major metabolite was butanedioic acid. There was a time-dependent increase in bacterial biomass, typical of bacterial growth, correlated with the decrease in pesticide concentration. The CO₂ production also increased significantly with the addition of pesticides to the bacterial growth media, demonstrating that, under aerobic conditions, the bacteria utilized endosulfan and malathion as a carbon source. Here, two bacterial strains are shown to metabolize two toxic pesticides into non-toxic intermediates.     

Isolation of mesotrione-degrading bacteria from aquatic environments in Brazil

Mesotrione is a benzoylcyclohexane-1,3-dione herbicide that inhibits 4-hydroxyphenyl pyruvate dioxygenase in target plants. Although it has been used since 2000, only a limited number of degrading microorganisms have been reported. Mesotrione-degrading bacteria were selected among strains isolated from Brazilian aquatic environments, located near corn fields treated with this herbicide. Pantoea ananatis was found to rapidly and completely degrade mesotrione. Mesotrione did not serve as a sole C, N, or S source for growth of P. ananatis, and mesotrione catabolism required glucose supplementation to minimal media. LC-MS/MS analyses indicated that mesotrione degradation produced intermediates other than 2-amino-4-methylsulfonyl benzoic acid or 4-methylsulfonyl-2-nitrobenzoic acid, two metabolites previously identified in a mesotrione-degrading Bacillus strain. Since P. ananatis rapidly degraded mesotrione, this strain might be useful for bioremediation purposes.     

Isolation and characterization of <Emphasis Type="Italic">Bradyrhizobium</Emphasis> sp. SR1 degrading two β-triketone herbicides

In this study, a bacterial strain able to use sulcotrione, a β-triketone herbicide, as sole source of carbon and energy was isolated from soil samples previously treated with this herbicide. Phylogenetic study based on16S rRNA gene sequence showed that the isolate has 100 % of similarity with several Bradyrhizobium and was accordingly designated as Bradyrhizobium sp. SR1. Plasmid profiling revealed the presence of a large plasmid (>50 kb) in SR1 not cured under nonselective conditions. Its transfer to Escherichia coli by electroporation failed to induce β-triketone degrading capacity, suggesting that degrading genes possibly located on this plasmid cannot be expressed in E. coli or that they are not plasmid borne. The evaluation of the SR1 ability to degrade various synthetic (mesotrione and tembotrione) and natural (leptospermone) triketones showed that this strain was also able to degrade mesotrione. Although SR1 was able to entirely dissipate both herbicides, degradation rate of sulcotrione was ten times higher than that of mesotrione, showing a greater affinity of degrading-enzyme system to sulcotrione. Degradation pathway of sulcotrione involved the formation of 2-chloro-4-mesylbenzoic acid (CMBA), previously identified in sulcotrione degradation, and of a new metabolite identified as hydroxy-sulcotrione. Mesotrione degradation pathway leads to the accumulation of 4-methylsulfonyl-2-nitrobenzoic acid (MNBA) and 2-amino-4 methylsulfonylbenzoic acid (AMBA), two well-known metabolites of this herbicide. Along with the dissipation of β-triketones, one could observe the decrease in 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibition, indicating that toxicity was due to parent molecules, and not to the formed metabolites. This is the first report of the isolation of bacterial strain able to transform two β-triketones.     

Study of metabolites from the degradation of polycyclic aromatic hydrocarbons (PAHs) by bacterial consortium enriched from mangrove sediments

The PAH metabolites produced during degradation of fluorene, phenanthrene and pyrene by a bacterial consortium enriched from mangrove sediments were analyzed using the on-fiber silylation solid-phase microextraction (SPME) combining with gas chromatography–mass spectrometry (GC–MS) method. Seventeen metabolites at trace levels were identified in different PAH degradation cultures based on the full scan mass spectra. In fluorene degradation cultures, 1-, 2-, 3- and 9-hydroxyfluorene, fluorenone, and phthalic acid were detected. In phenanthrene and pyrene degradation cultures, various common metabolites such as phenanthrene and pyrene dihydrodiols, mono-hydroxy phenanthrene, dihydroxy pyrene, lactone and 4-hydroxyphenanthrene, methyl ester, and phthalic acid were found. The detection of various common and novel metabolites demonstrates that SPME combining with GC–MS is a quick and convenient method for identification as well as monitoring the real time changes of metabolite concentrations throughout the degradation processes. The knowledge of PAH metabolic pathways and kinetics within indigenous bacterial consortium enriched from mangrove sediments contributes to enhance the bioremediation efficiency of PAH in real environment.     

Biodegradation of chlorsulfuron and metsulfuron-methyl by Aspergillus niger in laboratory conditions

Two sulfonylurea herbicides, chlorsulfuron and metsulfuron-methyl, were studied under laboratory conditions, in order to elucidate the biodegradation pathway operated by Aspergillus niger, a common soil fungus, which is often involved in the degradation of xenobiotics. HPLC-UV was used to study the kinetic of degradation, whereas LC-MS was used to identify the metabolites structure. In order to avoid the chemical degradation induced by a decrease in pH, due to the production of citric acid by the fungus, the experiments were performed in a buffered neutral medium. No significant degradation for both compounds was observed in mineral medium with 0.2% sodium acetate. On the contrary, in a rich medium, after 28 days the degradations, chemical degradation excluded, were about 30% for chlorsulfuron and 33% for metsulfuron-methyl. The main microbial metabolites were obtained via cleavage of the sulfonylurea bridge. In addition the fungus seems to be able to hydroxylate the aromatic ring of chlorsulfuron. In the case of metsulfuron-methyl the only detected metabolite was the triazine derivative, while the aromatic portion was completely degraded. Finally, the demethylation of the methoxy group on the triazine ring, previously observed with a Pseudomonas fluorescens strain, was not observed with A. niger.