优化毕赤酵母X33重组表达球形马拉色菌脂肪酶LIP1

Optimization of production of Malassezia globosa lipase LIP1 in recombinant Pichia pastoris X33

研究人工合成球形马拉色菌脂肪酶LIP1的基因,并成功在毕赤酵母X33中重组表达。利用响应面分析法,构建LIP1的酶活回归方程。由响应面分析,在温度27.0℃,pH值为7.69,酵母提取物浓度1.50%和葡萄糖浓度3.37%时,预测最大酶活为33U/mL。通过实验验证,最后结果为38U/mL。该值与模型预测基本一致。优化后的酶活提高了26.67%。通过响应面的条件优化,可以获得毕赤酵母X33重组表达LIP1的最大酶活。

Heterogeneous expression of Malassezia globosa LIP 1 gene in Pichia pastoris X33 system was studied. The effects of growth parameters on the LIP1 activity were investigate by response surface methodology. Based on ridge max analysis, the optimum LIP1 production conditions were obtained as follows： temperature 27.0℃, pH value 7.69, yeast extract 1.50% and glucose 3.37%. The predicted value of lipolytic activity was 33U/ml, and the actual value was 38U/ml, which was consistent with model prediction. Under optimized fermentation conditions, the lipolytic activity of recombinant LIP1 was 26.67%, which was higher than that of YPD medium.