基于响应面分析方法的有机磷水解酶全细胞活性表达条件的优化

Expression Condition Optimization of Whole Cell OPH Activity Based on Surface Response Analysis

为了提高有机磷水解酶(OPH)全细胞活性,对其表达条件进行了优化。首先采用Plackett-Burman试验设计对8个影响因素进行筛选,结果表明,诱导时间、IPTG浓度和诱导温度3个因素对OPH全细胞活性影响较大。在此基础上采用Box-Behnken设计,利用Design Expert软件进行二次回归分析,得到该基因工程菌的最佳表达条件为诱导温度18.86℃,IPTG浓度0.09mmol/L,诱导时间8.74h。OPH全细胞活性的最大预测值为27.829U/mL,试验验证值为27.8U/mL。

In order to obtain high expression of whole cell OPH activity,we optimized the expression conditions.Plackett-Burman design was undertaken to evaluate the effects of eight factors.With statistic regression analysis,the significant factors affecting whole cell OPH activity were determined as follows： induction time,IPTG concentration and induction temperature.Box-Behnken design was used to optimize these three critical internal factors,and determine the optimum levels and the relationships among these factors.By quadratic regression model equation with Design Expert statistic methods,the optimal levels of the variables were determined as： IPTG-induction temperature 18.86 ℃,IPTG concentration 0.09mmol/L and induction time 8.74h.Under this condition,the predicted whole cell OPH activity was 27.829U/mL,and the experimental whole cell OPH activity was 27.8U/mL.