邻氯硝基苯降解菌OCNB-1的分离鉴定与降解质粒  

Isolation,Identification and Degradative Plasmid Investigation of o-Chloronitrobenzene-Degrading Strain OCNB-1

作　　者：吴海珍[1] 何勤聪[2] 韦朝海[2] 王菊芳[1] 梁世中[1] 

机构地区：[1]华南理工大学生物科学与工程学院,广东广州510640 [2]华南理工大学环境科学与工程学院,广东广州510640

出　　处：《华南理工大学学报（自然科学版）》2008年第3期83-88,共6页Journal of South China University of Technology(Natural Science Edition)

基　　金：国家863计划项目(2006AA06Z378);国家自然科学基金资助项目(20777018);广东省科技计划项目(2007B03010311)

摘　　要：以邻氯硝基苯为唯一碳源、氮源和能源,从某化工厂废水处理站活性污泥中分离出一株细菌OCNB-1,经微生物自动测定仪WSVTK-R07.02和16S rDNA扩增测序初步鉴定为Pseudomonas putida;在培养基pH值为8.0、培养温度为32℃、摇床转速为120r/min的条件下,考察了该菌株降解邻氯硝基苯的能力,发现菌株42h内将起始浓度为1.07mmol/L的该化合物降解了约80%;经质粒检测,在OCNB-1菌株中发现一条质粒条带,选用7种内切酶对质粒pOCNB-1进行单酶切,得出质粒大小约为32 kbp;抗生素抗性实验表明菌株对红霉素、氨苄青霉素、青霉素钠的抗性与质粒无关,对利福平、氯霉素的抗性与质粒有关;通过细菌接合,质粒转移到无降解邻氯硝基苯能力的Pseudomonas.stutzeri受体菌中,接合子Pseudomonas.stutzeri能利用邻氯硝基苯为唯一碳源、氮源和能源,证明质粒为降解质粒.A bacterial strain OCNB-1 was isolated from the activated sludge in the wastewater treatment plant of a chemical factory, with o-chloronitrobenzene as the sole carbon, nitrogen and energy sources. The strain was then identified as Pseudomonas putida by the automatic biometrical system WSVTK-R07.02 and the 16S rDNA sequence analysis. Afterwards, the strain was applied to the degradation of 1.07 mmol/L o-chloronitrobenzene at a rotation speed of 120r/rain, a pH value of 8.0 and a temperature of 32℃ , finding that about 80% of o-chloronitrobenzene is degraded by the strain after 42 h. The results of plasmid detection indicate that there exists a plasmid strip in OCNB-1 strain, the plasmid size of which is proved to be about 32kbp according to the single cutting results by using 7 kinds of restriction enzymes. The results of antibiotic resistance reveal that the resistances of the strain to erythromycin, ampicillin and benzylpenicillin sodium have nothing to do with the plasmid, while those to rifampicin and chloramphenicol are related to the plasmid. Moreover, as the obtained plasmid can be transferred to Pseudomonas. stutzeri through bacterial conjugation and the conjugant can grow by using o-chloronitrobenzene as the sole carbon, nitrogen and energy sources, it is concluded that the plasmid carries the gene for o-chloronitrobenzene degradation.

关 键 词：邻氯硝基苯 降解 PSEUDOMONAS PUTIDA 鉴定 质粒 

分 类 号：X78[环境科学与工程—环境工程]