重组毕赤酵母发酵生产扩展青霉碱性脂肪酶的优化

Optimization of fermentation condition for Penicillium expansum alkaline lipase production using recombinant Pichia pastoris

通过毕赤酵母工程菌表达碱性脂肪酶,对表达条件进行了优化。先进行单因素实验,筛选出对碱性脂肪酶表达影响较大的4个因素:诱导时间、初始pH、甲醇添加量和接种量;在单因素实验的基础上设计四元线性回归实验,利用SPSS18数据处理软件进行数据优化,结果表明,诱导时间为96h、初始pH为6.0、甲醇添加量为0.5%、接种量为150mL时有最佳的表达效果,碱性脂肪酶活力为242.7U/mL。

In this study, the recombinant Pichia pastoris engineering bacteria was used to produce alkaline lipase （the engineering bacteria was constructed by our lab previously）, and expression condition was improved. At first single factor experiments were performed to select factors significant influence at expression, four factors was selected： inducing time, initialize pH, methanol concentration and inoculate. Four-liner regression with orthogonal design methods were constructed based on the single factor experiments, date was tested with SPSS 18 software. The results indicated that the optimization fermentation condition was that inducing 96 h, pH6.0, 0.5% methanol, inoculation 150 mL, the alkaline lipase activity was 242.7 U/mL.