重组肠激酶在甲醇酵母中高效表达的培养条件研究

The influence factors for the high-level expression of recombinant enterokinase in the Pichia pastoris

对重组肠激酶在甲醇酵母中高效表达的发酵条件进行了优化 ,并在 Bioflo 发酵罐中实现了高密度培养和高效表达 .结果表明 :温度 3 0℃ ,培养基 p H6.0 ,当酵母密度达到 2 0 0 A6 0 0 nm以上时 ,加入无水甲醇 ,控制甲醇的流加体积 ,使其终体积分数为 1 .0 %～ 3 .0 % ,继续诱导培养 72 h左右 ,酵母菌密度可达到 1 5 0 g/L,重组肠激酶总活性可达 5 8k

Enterokinase plays a key role in the construction of the expression system of artificial fusion protein because of its highly specific cleavage of the pentapeptide recognition sequence (Asp Asp Asp Asp Lys).In this paper,the influence factors for the high level expression of recombinant enterokinase in the Pichia pastoris yeast,such as the optimum temperature (30℃),pH 6.0,methanol volume concentration (1.0%~3.0%),yeast density and so on,were optimized in the Bioflo Ⅲ fermentor.The total enterokinase activity of 58 kIU/L culture broth and yeast dry weight of 150 g/L were obtained from experiments.