双表达解毒酶基因工程菌的酶活性分析及其应用  

Application and Activity Assay of Co-expressed Organophosphorus Hydrolase and Carboxylesterase(B1) Produced by Genetic Engineering Bacteria

作　　者：郑玉芝[1] 郭成君[2] 张建亮[1] 郭新民[2] 乔传令[1] 

机构地区：[1]中国科学院动物研究所,农业虫鼠害综合治理国家重点实验室,中国科学院研究生院,中国科学院研究生院北京100080 [2]中国人民大学环境学院,北京100872

出　　处：《食品科学》2006年第12期33-37,共5页Food Science

基　　金：国家"863"计划项目(2002AA601160;2005AA601020)

摘　　要：羧酸酯酶和有机磷水解酶可以水解不同的底物,为了获得降解谱广的解毒酶系,将两种酶共表达在同一载体。采用正交法对有机磷水解酶酶活性的分析条件进行了优化,以对硫磷为底物,底物浓度2×10-4mol/L、反应温度25℃、反应时间10min、pH8.0是测定酶活性最佳条件。在pH7.0～8.0的体系中存放一年活力可以保持50%以上。用于蚊子解毒实验,95%以上的对硫磷中毒蚊幼虫可以救活。在生物传感器等其它领域也都有应用。Carboxylesterase and organophosphorus hydrolase （OPH） are capable of hydrolyzing and detoxffying different pesticides. In order to get the widest substrate spectrum organophosphorus hydrolase and carboxylestcrase （B1） were coexpressed in one genetic engineering bacteria. The activity of OPH was assayed by orthogonal test, and the optimum condition were substrate concentration 2 × 10^-4mool/L, temperature 25℃, reaction time 10min and pH8.0. In the detoxification of parathion on mosquitoes, 95% of the mosquitoes can be survived. And it has been used in biosensor for pesticide detection.

关 键 词：羧酸酯酶 有机磷水解酶 酶活性测定 

分 类 号：Q78[生物学—分子生物学]