甲基对硫磷水解酶基因的高效表达及酶活性分析  

Heterogenous Gene Expression of Methyl Parathion Hydrolase and Analysis of the Enzyme Activity

作　　者：刘璐璐[1] 周亚凤[1] 张治平[1] 刘虹[1] 张先恩[1] 

机构地区：[1]中国科学院武汉病毒研究所,武汉430071

出　　处：《微生物学通报》2004年第6期77-82,共6页Microbiology China

摘　　要：假单胞菌WBC 3的甲基对硫磷水解酶基因mph在大肠杆菌系统AD494(DE3) pET32a ( + )中实现了高效可溶性融合表达。表达的重组酶能够有效地被亲合层析纯化。酶学性质分析表明 ,重组酶对底物乙基对硫磷水解能力较野生酶相比有近 4倍的提高 ,对甲基对硫磷和杀螟松的水解活力无明显变化。以融合蛋白形式存在的重组酶在室温及Methyl parathion hydrolase (MPH, E.C.3.1.8.1) coding gene mph from Pseudomonas sp. WBC-3, isolated and identified by our lab, was successfully expressed in E. coli AD494 (DE3)/ pET32a(+) system as soluble fusion form at high level. The recombinant MPH showed nearly 4～5 fold higher specific activity to parathion than the enzyme from Pseudomonas sp. WBC-3. In addition, the thermal stability of the recombinant enzyme was improved comparing with the wild type enzyme.

关 键 词：有机磷水解酶 硫氧还蛋白 融合表达 

分 类 号：Q786[生物学—分子生物学]