重组SOD在毕赤酵母中表达的发酵工艺研究

Research on the fermentation technology of recombinant SOD expression in Pichia pastoris

以实验室构建的产SOD重组菌为出发菌株,研究其摇瓶发酵工艺条件以提高表达水平,进而在5 L高密度发酵.以确定接种后诱导时间(16 h)、诱导剂浓度(2%)、温度(30℃)、初始pH值(6.2)和诱导时间72 h为条件,摇瓶水平酶活比初始时提高了64%;经5 L罐发酵实验,酶活达到40 864 U·mL-1,是摇瓶的86倍.

In this article, study fermentation conditions of the recombinant strains in shake culture, aiming at improving the protein expression and fermenting in 5 L fermentation tank. The result indi- cates that the suitable fermentation conditions for the recombinant cell are as below： the temperature of cell growth and protein expression is 30 ℃, the methanol feeding is 2% （volume ratio of the medium） per day, the initial pH is 6.2, the induction time is 72 h. According to these conditions, the enzyme activity of shake culture is increased by 64% comparing with before optimization. In 5 L fermentation cultivate, the enzyme activity reaches 40 864 U. mL-1, 86 times of the shake culture.