重组人抗凝血酶发酵条件的优化

Research on Optimum Fermentation Condition of Recombined Human Antithrombin

目的:通过实验室发酵条件优化工作,实现在巴斯德毕赤酵母中高效表达重组人抗凝血酶。方法:在摇瓶培养条件下,应用正交实验方法考查人抗凝血酶重组毕赤酵母菌pPIC9K-AT-02的培养温度、培养基pH值、接种比例、甲醇补加间隔时间及甲醇补加浓度等5种因素对重组人抗凝血酶活性的影响,确定最优发酵条件。结果与结论:筛选出的最终发酵条件为培养温度30℃、培养基pH6.0、接种比例20%、甲醇补加间隔时间24 h、甲醇补加浓度2%,重组人抗凝血酶在巴斯德毕赤酵母中表达活性为4098 U/L,比原始活性提高了150%。

Objective： To express antithrombin gene in Pichia pastoris by the optimum fermentation condition.Methods： The influence of temperature,culture medium pH,inoculation ratio,the time and concentration of methanol addition were studied to optimize fermentation condition of pPIC9K-AT-02 by orthogonal experiment.Results Conclusion： The optimum fermentation conditions of recombined P.pastoris were as follows： 30℃ culti-vation temperature,using culture medium pH6.0,inoculation ratio 20%,adding 2% methanol every 24 h.The opti-mum fermentation condition have determined to express antithrombin highly and its activity have achieved 4098 U / L and increased to 150% than original activity.