摘要
采用Plackett-Burman设计法对影响巴斯德毕赤酵母(Pichia pastoris)表达重组人血清白蛋白-粒细胞集落刺激因子突变体(rmHSA-GCSF)的主要参数进行了摇瓶试验,选取到3种显著效应的因素:诱导pH、甲醇添加浓度和诱导时间。再利用响应曲面法(Response surface method)对这3种因素进行进一步研究,得到最佳培养条件:甲醇诱导浓度为2.45%,诱导pH为5.54,诱导时间为66.4 h。采用此优化参数,rm HSA-GCSF表达量的最大预测值达157.5 mg/L。此后,根据优化的培养条件,进行150 L发酵罐放大试验,rm HSA-GCSF表达量达到600 mg/L左右,成功实现rm HSA-GCSF的高效表达。
A Plackett-Burman design was used to evaluate the factors related to rmHSA-GCSF expression by pichiapastoris in shake flask. Induction pH, methanol concentration and induction time with significant influence on rmHSA-GCSF fermentation were selected. The response surface methodology was used to obtain the optimal region of the three factors. The optimal process condi- tions were induction pH 5.54, methanol concentration 2.45% and induction time 66.4 h. Under these conditions, the maximum predicted yield of rmHSA-GCSF was 157.Smg/L. According to the optimization of culture conditions in shake flask, the rmHSA-GCSF yield was increased up to about 600 mg/L in 150 L fermentor. High-level expression of rmHSA-GCSF was successfully achieved.
出处
《发酵科技通讯》
CAS
2015年第2期29-33,共5页
Bulletin of Fermentation Science and Technology
