菊酯类农药降解酶基因原核表达条件的优化

Optimization of prokaryotic expression conditions of pyrethroid degrading enzyme gene

绘制了菊酯类农药降解酶基因工程菌E.coliBL21(DE3)/pET-28a-est825的生长曲线,优化了诱导起始菌液浓度、诱导剂浓度、诱导温度及诱导时间等条件,得到该基因工程菌诱导表达的最适条件:发酵培养至菌液OD600值为4时,添加终浓度为6g/L的乳糖,37℃诱导8h,酶的表达量最高,酶活为95U/mL,纯化后的重组酶分子量为30.1ku。

In this study,the growth curve of gene engineering bacteria E.coli BL21（DE3）/pET-28a-est825 of pyrethroid degrading enzyme was drawn,and initial induction culture OD600,inducer concentration,induction temperature and induction time were optimized.The results showed that the optimal induction culture OD600 was 4,the optimal inducer concentration was 6 g/L,the optimal induction temperature was 37℃,and the optimal induction time was 8 h.Under the optimal conditions,the enzyme activity reached 95 U/mL.The molecular mass of recombinant enzyme was 30.1 ku.