摘要
合成opdA基因,构建表达载体pET-28b-opdA,将其转化至大肠杆菌BL21(DE3)中得到重组工程菌株,并对重组菌株的诱导表达条件进行优化。结果表明:诱导前菌液OD_(600)的吸光值为0.5,IPTG诱导浓度为0.1mmol/L,诱导温度为37℃,诱导时间为5 h。opdA酶对甲基对硫磷的K_m为42.6μmol/L,其ν_max为32.669μmol/(L·min),比酶活为24.48U/mg。
The synthetic gene opdA was inserted into vector pET-28b to construct a recombinant plasmid pET-28b-opdA, which .was subsequently transformed into BL21 （DE3）, producing a recombi- nant engineering strain BL21 （DE3）/pET-28b-opdA. And the recombinant strain ＇s induction and ex- pression conditions were optimized. The results showed that the OD600 before induction was 0.5, and the induction was made at 0. 1 mmol/L IPTG and 37℃ for 5 h. The Km of opdA enzyme for parathion-methyl was 42.6 μmol/L,the maximum reaction velocity（ Vmax）was 32. 669 μmol/（L.min）, and the specific activity of opdA enzyme was 24.48 U/mg.
出处
《上海农业学报》
CSCD
北大核心
2013年第5期1-5,共5页
Acta Agriculturae Shanghai
基金
上海市科委基础重点项目(10JC1413800)资助
关键词
有机磷降解酶
基因工程
蛋白表达
蛋白纯化
酶活性
Organophosphate-degrading enzyme
Gene engineering
Protein expression
Protein purification） Enzyme activity
