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 surfacemethod）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，rmHSA-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，rmHSA-GCSF 表达量达到600 mg/L左右，成功实现rmHSA-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 conditions 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 157.5mg/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.