响应面法优化重组毕赤酵母表达米曲霉碱性蛋白酶诱导条件的研究

为探索重组米曲霉碱性蛋白酶（rAlp）在毕赤酵母中表达的最佳诱导条件，本研究在单因子实验的基础上，运用Box-Behnken设计的响应面试验对诱导条件进行了优化。根据回归分析确定了影响rAlp表达的因子，求得最佳诱导条件为：诱导温度28．53℃，诱导pH6．63，甲醇浓度1．28％。此工艺条件下碱性蛋白酶活力实测值为49．78U／mL，回归模型的预测值与实测值的相对误差〈1％，说明该回归方程与实际情况拟合很好。

Studies on application of response surface method for inducing conditions of expression of recombinant Aspergillus oryzae alkaline protease in Pichia pastoris

In order to optimize the conditions for expressing recombinant Aspergillus oryzae alkaline protease （rAlp） induced by methanol, the response surface methodology （RSM） designed by Box-Behnken was applied based on single factor tests. The factors relating to the expression of rAlp were obtained with method of regression analysis. The optimum inducing conditions were as follows： temperature of 28.53℃, pH 6.63, methanol concentration of 1.28%. The actual activity of alkaline protease was 49.78 U/mL under the above conditions. The relative error was 〈 1% between actual protease activity and the activity calculated according to the regression model, which suggested that regression equation fitted well with actual condition.