响应面法优化纤维堆囊菌So ce 90原生质体的制备条件

Optimization of protoplast formation of Polyangium cellulosum So ce 90 using response surface methodology

采用响应面法优化纤维堆囊菌So ce 90的原生质体制备条件。选择溶菌酶浓度,酶解时间和酶解温度作为考察因子,在单因素试验的基础上,采用Box-Benhnken中心组合设计方法进行三因素三水平试验设计。以原生质体生成率作为响应值,进行多元二次响应回归分析,得出纤维堆囊菌So ce 90原生质体制备的最佳条件为:溶菌酶浓度1.40 mg/mL,酶解时间为160min,酶解温度为30℃,响应模型预测的原生质体最高生成率为87.78%,在最优的条件下进行验证试验,得到原生质体生成率为86.23%,与预测值间的相对误差为1.76%,表明采用响应面法优化纤维堆囊菌So ce 90原生质体的制备条件是可行的。

Response surface methodology （RSM） was applied to optimize the protoplast preparation conditions from Polyangium cellulosum So ce 90. The concentration of lysozyme, enzymatic time and enzymatic temperature were selected as casual factors. Base on the single factor experiments, a three-factor-three-level experiment design was developed by Box-Benhnken central composite design method. The protoplast formation rate was used as the response values and RSM was applied to analysis the results. The optimum protoplast preparation conditions from Polyangium ceUulosum So ce 90 obtained by RSM analysis were as follows： the concentration of lysozyme 1.40 mg/mL, enzymatic time 160 rain and enzymatic temperature 30℃. The best protoplast formation rate obtained by RSM analysis was 87.79 %. The protoplast formation rate in a validation experiment was 86.23 %. The relative error between predictive values and experiment values was 1.77 %. These results demonstrated that the method was feasible.