转解毒酶基因工程菌的构建及酶活性分析

Construction of Detoxifying Gene Recombinant Bacterium and Enzyme Activity Analysis

实验构建了能高效降解有机磷、有机氯等四大类农药的转解毒酶基因工程菌。将抗性库蚊解毒酶酯酶B1基因片段引入融合表达载体pThioHisA中,转化入大肠杆菌DH5α,在IPTG诱导下,经过8 h,酯酶B1在大肠杆菌中的获得融合高效表达。研究了工程菌的酶酯活性,重组质粒pThioHisA-B1表达的酯酶融合蛋白具有较高的酯酶B1活性,能高效降解酯酶的特异性底物α-乙酸萘酯(-αNA)和β-乙酸萘酯(-βNA),该工程菌将为农药污染的生物治理提供新手段。

A detoxifying gene esterase B1 was cloned into fusion expression vector pThioHisA. The recombinant vector pThioHisA-B1 was constructed and transformed into E. coli DH5a. High level of expression of the target protein was obtained 8 hours after induced by IPTG. The protein expressed by pThioHisA-B1 possessed high esterase activity and it can degrade the specific substrate α-NA and β-NA of esterase effectively. A novel engineering bacterium containing recombinant esterase B1 gene was constructed that can degrade four kinds of insecticides including organophosphate and organochlorine. The use of the engineered bacterium could provide a novel way in bioremediation of pesticide contamination.