甲基对硫磷降解菌假单胞菌WBC-3的筛选及其降解性能的研究  

Study on Pseudomonas sp. WBC-3 Capable of Complete Degradation of Methylparathion

作　　者：陈亚丽[1] 张先恩[1] 刘虹[1] 王银善[1] 夏祥明[1] 

机构地区：[1]中国科学院武汉病毒研究所,武汉430071

出　　处：《微生物学报》2002年第4期490-497,共8页Acta Microbiologica Sinica

基　　金：国家"86 3"生物技术项目 (SZ 0 3 0 1 0 3)

摘　　要：从农药污染土样中分离出的一株细菌具有彻底降解甲基对硫磷的能力。该菌经生理生化特性分析和 1 6SrDNA序列同源性分析 ,鉴定为假单胞菌属 ,命名为Pseudomonassp .WBC 3。该菌在pH7～ 8、温度 2 3℃～ 3 0℃范围内均生长良好 ,对甲基对硫磷的耐受浓度在单纯无机盐培养基中可达到 80 0mg L ,在含有 0 1 %葡萄糖的培养基中可达到 2 0 0 0mg L。该菌能够以甲基对硫磷作为唯一碳源、氮源 ,将其彻底降解作为生长基质 ,对于 3 0 0mg L甲基对硫磷的降解速度达 1 5mg L·h ,于 2 2h后达到其稳定生长期。该菌对于多种有机磷农药及部分芳烃类化合物具有生化代谢能力。从该菌的细胞周质组分中纯化出的有机磷水解酶在SDS PAGE胶上显示为分子量约为 3 3 5× 1 0 3 的条带。A bacterial strain was isolated from the polluted soil around Shashi Pesticides Factory, which was capable of complete degradation of methylparathion. Through chemotaxonomic characterizations and phylogenetic inference based on 16S rDNA sequence analysis, the strain was identified as a member of the genus and was named as \%Pseudomonas\% sp. WBC\|3. It can tolerate high\|concentration methylparathion up to 800mg/L in basic medium and up to 2000mg/L in 0\^1% glucose medium. Using 300mg/L methylparathion as its sole carbon and nitrogen sources, the strain was able to degrade 15 mg of parathion per liter per hour and reached its stationary phase in about 22 hours. The strain possessed broad\|spectrum degradative capability to kinds of organophosphorus pesticides and aromatic compounds. Its organophosphate hydrolase was purified from the periplasm of WBC\|3 to homogeneity, through a whole process consisting of ammonium sulfate precipitation, Sephadex G\|75 gel filtration, Q Sepharose FF ionexchange column chromatography. The purified enzyme showed a single band on SDS\|PAGE gel with an approximate molecular weight of 33\^5 kD.

关 键 词：甲基对硫磷降解菌 假单胞菌 WBC-3 筛选 降解性能 有机磷水解酶 农药 

分 类 号：Q939[生物学—微生物学]