在大肠杆菌中表达重组CotA蛋白的培养条件优化

Optimization of Culture Conditions of Recombinant CotA Protein Expressed in Colon Bacillus

为了提高大肠杆菌对重组CotA蛋白的表达量，对本实验室构建的含CotA基因表达载体的工程菌株的培养条件进行优化。通过单因素试验及正交试验L18（3 7）优化了产酶的最佳培养条件为：摇床转速为180r／min、装液量为100mL／500mL锥形瓶、接种量为5％、诱导温度25℃、诱导剂IPTG浓度为1．0mmol／L、诱导时间为12h，以及加诱导剂时菌体OD600值为1．0，其中诱导剂IPTG浓度和菌体OD600值对CotA蛋白表达影响显著。

Objective： This study is to optimize the culture conditions of recombinant Escherichia. eoli BL21（DE3）- pET-22b（＋）-CotA which is available to express the CotA protein. Through single factor and orthogonal design L18 （3 7）experiments to make sure of the optimum conditions for culture conditions. The result shows that 180 r/min of rotational speed, 100 mL/500 mL of Erlenmeycr flask, 5% of inoculum volume, temperature 25℃, 1.0 mmol/L IPTG, cell density OD660 of 1.0, cultivation 12 h are the optimum conditions. The concentration of IPTG and cell are the induced expression conditions which has significant influence on the CotA protein.