苯系物降解菌Pseudomonas putida SW-3的筛选及其降解苯的研究

【目的】以苯、甲苯和苯乙烯为唯一碳源,从工业石油废水中筛选苯系物降解菌,分析其降解特性,探讨底物间相互作用对降解情况的影响。【方法】经生理生化和16S r RNA基因分析进行菌种鉴定,采用顶空气相色谱法测定苯系物含量,通过细胞的疏水性、乳化能力、排油圈及透射电镜观察分析菌株降解特性。【结果】经鉴定该菌为Pseudomonas putida,命名为SW-3菌株。最适降解条件下,单位菌体对苯、甲苯和苯乙烯的最大降解速率分别为0.072、0.035和0.019 g/(L·h),苯系混合物的总降解率达79.99%。底物降解实验表明,苯可促进甲苯和苯乙烯的降解,而苯乙烯则能抑制甲苯的降解。菌株的吸附、摄取和降解特性的研究发现,菌株SW-3在自身分泌的表面活性剂的协助下以耗能的方式运输苯。【结论】菌株SW-3具有产生表面活性剂和降解苯系物的能力,且底物间的相互作用能够显著影响菌株对不同底物的降解。     

一株海草沉积物菲降解菌的筛选、鉴定和降解特性

【背景】多环芳烃(Polycyclic Aromatic Hydrocarbons,PAHs)是一类高毒性的有机污染物,在海洋环境尤其是沿海环境中广泛分布。海草床生态系统作为沿海环境的重要组成部分,深受环境污染等人类活动的影响而处于严重衰退的状态。微生物修复是修复环境中多环芳烃污染的重要途径,具有经济简便、环境友好和无二次污染等特点。【目的】从深圳市大亚湾的海草床沉积物中筛选获得高效多环芳烃降解菌,并分析其降解特性,从而探究海草床生态系统中多环芳烃污染物的微生物修复可行性。【方法】以多环芳烃菲为唯一碳源从海草床沉积物样品中筛选菌株,再通过形态学观察、生理生化实验和16SrRNA基因序列对筛选的菌株进行鉴定,并利用特定引物扩增多环芳烃降解的功能基因——双加氧酶(nidA)基因,最后通过培养实验分析该菌株对菲的降解特性。【结果】筛选出一株高效降解菲的菌株SCSIO 43702,经鉴定为玫瑰杆菌属(Roseovarius)的潜在新菌,并成功扩增得到双加氧酶相似(nidA like)基因;培养实验结果表明,玫瑰杆菌SCSIO 43702在10 d内对100 mg/L菲的降解率最高可达96%,而且其对菲的最适降解条件为:温度30°C、pH值7.5和8.0、盐度3%。【结论】玫瑰杆菌SCSIO 43702凭借其良好的菲降解能力和较强的环境适应性,具有进一步被开发为微生物菌剂以用于多环芳烃污染修复的巨大潜力,为海草床生态系统中多环芳烃污染的微生物修复研究提供了理论依据和可利用的微生物资源。     

一株苯酚降解菌的筛选、鉴定及其降解特性

本研究采用逐量分批驯化的方法,从造纸废水中分离得到一株能够以苯酚为唯一碳源生长的苯酚降解菌株F5-1.经形态观察、生理生化特性鉴定及16S rDNA序列分析,将该菌株鉴定为克雷伯菌(Klebsie-lla sp.).该菌株能够在7 h时完全降解初始浓度为100 mg/L的苯酚,降解苯酚主要发生在生长对数期;在pH 5.0～9.0,NaCl浓度0～80 g/L,温度20～40℃范围内,菌株F5-1均可有效降解初始浓度为100～1 200 mg/L的苯酚;能够耐受的最大苯酚浓度为1 500 mg/L.本研究结果表明,F5-1菌株对处理环境条件复杂的含酚废水具有潜在的应用前景.     

一株苯系物降解真菌筛选及降解特性

采用逐量分批驯化的方法以污水处理厂污泥作为菌源,苯、甲苯、二甲苯为唯一碳源,驯化、分离、筛选能够有效降解苯系物的真菌,命名为B1。采用单因素以及正交实验方法并对真菌降解环境影响因素及降解效率进行了测定和研究。结果表明:真菌B1对苯系物降解的最佳条件为C:N=5:1,pH5,温度30℃,菌种接种量为5.5ml(50ml培养基)。采用GC对初始液相浓度0~90mg/L范围内的苯系物降解效果进行测定,未发现苯系物对真菌降解活性产生抑制作用。真菌对苯系物的降解效率为:甲苯>苯>二甲苯,最高降解效率分别达到87.39%,85.21%,81.47%。混合物降解效果略高于单一底物的降解效果。     

一株聚乳酸降解菌的筛选、鉴定及其降解特性

【目的】分离可有效降解聚乳酸(polylactic acid,PLA)的好氧细菌,并研究其生长特性和降解特性,为环境中PLA废弃物的生物修复提供依据。【方法】通过16S rRNA基因序列分析对所筛选菌株进行分子生物学鉴定,采用扫描电镜和傅里叶变换红外光谱(Fourier transform infrared spectroscopy,FTIR)分析降解前后PLA膜的形貌和化学结构变化。【结果】从活性污泥中筛选获得一株芽孢杆菌(Bacillus sp.) JA-4,30 d后PLA失重率可达10.6%。pH 8.0、30℃以及接种量20%的条件下,7 d后PLA的失重率可达到5.6%。明胶对PLA的生物降解具有显著促进作用,当明胶浓度为3%时,降解10 d后PLA的失重率达到23.1%,降解速率也大大提高。FTIR分析表明该菌株通过水解酯键来实现PLA的降解。【结论】本研究为PLA的生物降解提供了新的微生物资源,为环境中PLA废弃物的有效降解提供技术支持。     

氯苯降解菌的筛选鉴定及降解特性研究

本文采集化工厂排污口的污泥样品, 在含有氯苯为唯一碳源的基本培养基中, 先后分离筛选出7株能够降解氯苯的微生物菌株。通过对分离菌株的16S rRNA基因序列进行分析, 发现其中5株细菌分别属于放线菌目的考克氏菌属(KD139)、红球菌属(KD140和KD142)和节杆菌属(KD230和KD232), 1株细菌属于杆菌目的芽胞杆菌d属(KD178), 另外1株细菌属于黄色单孢菌目的寡食单胞菌属(KD237); 同时我们构建了系统进化树, 确定分离菌株的相对进化地位。本文还利用气相色谱方法, 对分离菌株降解氯苯的能力进行了初步分析, 其中寡食单胞菌KD237降解氯苯能力最高, 24 h内氯苯分解率达60.78%。     

一株芘降解菌的分离鉴定及其降解效果

以芘为唯一碳源,采用平板升华法,从徐州市卧牛山焦化厂周围污染土壤中分离得到一株芘降解菌SE12.经形态观察、生理生化试验和16S rDNA鉴定,该菌株属于分枝杆菌属(Mycobacterium sp.)菌株,与快速生长型非致病性南非分枝杆菌(M.austroa fricanum ATCC33464)的同源性达到98%.SE12降解芘的最适pH和温度为pH9和30℃.当土壤芘初始含量为100和200mg.kg-1,SE12接种量为107CFU.g-1时,30℃培养28d后土壤芘降解率分别达到97%和99%.利用双加氧酶基因的同源序列引物nidAF/nidAR和nidBF/nidBR进行扩增,得出了该菌株编码双加氧酶大亚基和小亚基的基因片段,它们与已知降解芘的分枝杆菌的双加氧酶基因具有高度同源性.     

苯扎贝特降解菌的筛选及降解特性

【目的】药品苯扎贝特在水环境中频繁检测出,对环境的潜在危害不容忽视。我们筛选分离降解苯扎贝特的细菌,并研究其降解特性。【方法】根据分离菌株的细胞形态结构、生理生化特征及其16S rRNA基因序列分析鉴定降解菌,高效液相色谱法测定苯扎贝特,以判定该菌株的降解能力。【结果】分离菌株B-31属恶臭假单胞菌(Pseudomonas putida),降解机制是共代谢。降解最佳条件为30℃、pH7。此条件下,以1%甲醇为初级基质,30mg/L苯扎贝特的5日降解率为48%。当分别以5g/L葡萄糖、蛋白胨、酵母粉为初级基质时,可使降解率提高到61%,、72.6%、76.67%。【结论】这是国内首次报道恶臭假单胞菌可以通过共代谢降解苯扎贝特,该研究为利用细菌发酵消除水环境中苯扎贝特污染提供基础。     

一株PET降解菌株的筛选鉴定及降解特性

本研究进行了聚对苯二甲酸乙二醇酯(polyethyleneterephthalate,PET)降解菌株的分离、筛选和鉴定,以及降解机制的探究.用"分级筛选"策略,先利用塑料类似物对苯二酸二甲酯(diethyl terephthalate,DET)进行富集培养,在以PET颗粒为唯一营养源的无机盐固体培养基上进行涂布,从垃圾填埋场PET塑料样品中筛选获得具有降解PET颗粒能力的菌株JWG-G2.通过菌株形态观察、生理生化特性及16S rRNA序列分析,鉴定该菌株属于微杆菌属(Microbacterium sp.).菌株JWG-G2在pH 7.0、30℃时生长状态最佳.经菌株JWG-G2处理后,PET颗粒表面酯键特征官能团明显减少;PET颗粒失重率达到1％.菌株JWG-G2能够显著降解PET中间体对苯二酸单羟乙酯钠盐(monohydroxyethyl terephthalate,MHET)和对苯二甲酸双羟乙酯(bishydro-xyethyl terephthalate,BHET)多聚体,其降解率分别为4.5％和11.2％.菌株JWG-G2具有较好的PET颗粒及其中间体降解作用,为降解机制的深入研究提供一定理论基础.     

降解吡啶复合菌系MD1的筛选、降解特性及代谢途径

【目的】为筛选吡啶高效降解复合菌系,促进高浓度吡啶废水的降解。本研究围绕吡啶降解复合菌系的筛选、降解特性及代谢途径,旨在获得吡啶高效降解复合菌系,为高浓度吡啶废水微生物降解及完全矿化提供理论依据和技术支撑。【方法】以吡啶为唯一碳氮源从某农药废水处理系统好氧活性污泥中筛选得到一个吡啶高效降解复合菌系MD1。采用16S rRNA高通量测序技术探究了MD1的群落结构及多样性,通过单因素实验考察了MD1的降解特性,利用气相色谱-质谱联用仪对MD1降解吡啶的代谢产物进行了初步检测与鉴定,推测吡啶可能的降解途径。【结果】结果显示,在温度30 ℃、pH 8.0、NaCl浓度0.1%的最佳条件下培养72 h,MD1对初始浓度1 400 mg/L的吡啶降解率为98.44%±0.27%。在属水平上,MD1主要由副球菌属(Paracoccus sp.)、布鲁氏菌属(unclassified_Brucellaceae)、无色杆菌属(Achromobacter sp.)等组成。由代谢产物检测结果初步推测MD1对吡啶的代谢途径为吡啶→烟酸→6-羟基烟酸→2,5-二羟基吡啶→N-甲酰基马来酰胺酸→马来酰胺酸→马来酸→CO₂+H₂O。【结论】研究筛选得到一个可高效降解吡啶、降解性能稳定的复合菌系MD1。解析了MD1的微生物组成多样性和群落结构,推测了MD1可能的代谢途径,研究结果丰富了吡啶降解微生物资源。     

Is interspecies hydrogen transfer needed for toluene degradation under sulfate-reducing conditions?

Sediments from a hydrocarbon-contaminated aquifer, where periodic shifts between sulfate reduction and methanogenesis occurred, were examined to determine whether the degradation of toluene under sulfate-reducing conditions depended on interspecies hydrogen transfer. Toluene degradation under sulfate-reducing conditions was inhibited by the addition of 5 mM sodium molybdate, but the activity was not restored upon the addition of an actively growing, hydrogen-using methanogen. Toluene degradation was not inhibited in microcosms where hydrogen levels were maintained at a level theoretically sufficient to inhibit toluene degradation if the process proceeded via interspecies hydrogen transfer. Finally, the addition of carbon monoxide, a potent inhibitor of hydrogenase activity, inhibited hydrogen but not toluene consumption in sulfate-reducing microcosms. These results suggest that toluene is degraded directly by sulfate-reducing bacteria without the involvement of interspecies hydrogen transfer. The sequence of experiments used to reach this conclusion could be applied to determine the role of interspecies hydrogen transfer in the degradation of a variety of compounds in different environments or under different terminal electron-accepting conditions.     

Simultaneous determination of benzene, toluene, ethylbenzene, and xylenes in urine by thermal desorption–gas chromatography

The determination of metabolites of benzene, toluene, ethylbenzene, and xylenes in urine has been used to assess human exposure to these compounds. The analyses of urine samples for these metabolites are tedious and time consuming. The determination of unmetabolized individual compounds in urine has been studied previously with some success. A simultaneous determination of several unmetabolized VOC compounds in urine by thermal desorption–gas chromatography was conducted to assess the exposure of smokers and nonsmokers to these compounds. The method of thermal desorption–GC was sensitive enough to detect a significant difference in exposure levels due to the contribution of light smoking in the environmentally-exposed group.     

Isolation and characterization of three <Emphasis Type="Italic">Sphingobium</Emphasis> sp. strains capable of degrading isoproturon and cloning of the catechol 1,2-dioxygenase gene from these strains

Three strains of bacteria (designated as YBL1, YBL2, YBL3 respectively) capable of degrading isoproturon, 3-(4-isopropylphenyl)-1, 1-dimethylurea, were isolated from the soils of two herbicide plants. Based on the comparative analysis of the 16S rRNA gene, and phenotypic and biochemical characterization, these strains were identified as Sphingobium sp. The optimum conditions for isoproturon degradation by these strains were pH 7.0, and temperature 30°C. Mg²⁺ (1 mM) enhanced the isoproturon degradation rate, while Ni²⁺ and Cu²⁺ (1 mmol l⁻¹) inhibited isoproturon degradation significantly. These three strains also showed the ability to remove the residues of other phenylurea herbicides such as chlorotoluron, diuron and fluometuron in mineral salt culture medium. The N-demethylation was the first step of degradation of dimethylurea-substituted herbicides. Strain YBL1 was found capable of degrading both dimethylurea-substituted herbicides and methoxymethylphenyl-urea herbicides i.e. linuron (3-(3,4-dichlorophenyl)-1-methoxy-1-methylurea). Using the PCR method, partial sequences of the catechol 1,2-dioxygenase gene were obtained from these strains.     

Utilization of toluene and xylenes by a nitrate-reducing strain of Pseudomonas maltophilia under low oxygen and anoxic conditions

Abstract A nitrate-reducing strain of Pseudomonas maltophilia isolated from sewage sludge degrades toluene, and at least two isomers of mixed xylenes, either in the presence of 2% oxygen or under anoxic conditions when nitrate is present. When individual isomers of xylene are provided only meta and para -xylene are utilized. When mixed xylenes are provided all three isomers may be utilized. In cultures limited by electron acceptor availability, succinate, when present as the major carbon source, does not prevent hydrocarbon utilization. Toluene and xylenes continued to be utilized either with limiting nitrate alone, or with limiting nitrate and oxygen present simultaneously when a hundred-fold excess of succinate is present in the medium. The data suggest that in groundwater containing low levels of oxygen and nitrate, or nitrate only as the electron acceptor, aromatic hydrocarbons may continue to be utilized even in the presence of an excess of readily-degradable non-hydrocarbon organic substrates. These data have implications for bioremediation studies. The strain of Pseudomonas maltophilia used in this study does not degrade benzene, and the presence of benzene does not affect toluene utilization.     

The role of active-site residues in naphthalene dioxygenase

The three-component naphthalene dioxygenase enzyme system catalyzes the first step in the degradation of naphthalene by Pseudomonas sp. strain NCIB 9816-4. A member of a large family of bacterial Rieske non-heme iron oxygenases, naphthalene dioxygenase is known to oxidize over 60 different aromatic compounds, and many of the products are enantiomerically pure. The crystal structure of the oxygenase component revealed the enzyme to be an α₃β₃ hexamer and identified the amino acids located near the active site. Site-directed mutagenesis studies have identified the residues involved in electron transfer and those responsible for controlling the regioselectivity and enantioselectivity of the enzyme. The results of these studies suggest that naphthalene dioxygenase can be engineered to catalyze a new and extended range of useful reactions.