基于蛋白酶活性控制的重组人β干扰素毕赤酵母表达体系优化  

Optimization of Pichia pastoris Expression System Related to Recombinant Human Interferon β Based on Control of Protease Activity

作　　者：张旦旦 窦文芳 许正宏 史劲松 

机构地区：[1]江南大学医药学院,江苏无锡214122

出　　处：《中国医药工业杂志》2012年第9期743-747,共5页Chinese Journal of Pharmaceuticals

基　　金：国家"重大新药创制"科技重大专项(2009ZX09102-221);国家自然基金(31000016);江苏省自然科学基金(企业博士创新项目BK2009518);江苏省科技支撑计划(BE2009629)

摘　　要：以毕赤酵母人β干扰素表达体系为研究对象,考察与蛋白酶活性相关的营养和环境因素对外源蛋白hIFN-HAS表达/降解的影响。结果表明,pH、谷氨酸和EDTA是显著影响hIFN-HAS表达的3个因素。通过响应面分析得到hIFNβ-HAS表达的最优条件为pH 5.5、谷氨酸0.18%、EDTA 12 mmol/L,目的蛋白表达量为24.46 mg/L。发酵终产物的电泳结果表明:条件优化后,hIFNβ-HSA降解带明显变淡,说明营养和环境因素的优化一定程度上抑制了蛋白酶的活性,由此增加了目的蛋白的积累。The effect of nutrition and environmental factors related to protease activity on the expression and degradation of exogenous protein （IFN β-HAS） was investigated in Pichia pastoris expression system. The results suggested that pH, glutamic acid and EDTA were three key factors significantly affected the expression of IFN β-HAS. Moreover, the optimal condition was determined as pH 5.5, glutamic acid 0.18% and EDTA 12 mmol/L by response surface analysis of three factors and three levels. The production of IFN β-HSA reached 24.46 mg/L. The electrophoresis results of the final fermentation product showed that the degradation of IFNβ-HSA reduced significantly and the production of target protein increased due to the activity of protease was inhibited after the optimization of nutrition and environmental factors.

关 键 词：毕赤酵母 Β干扰素 融合蛋白 响应面分析 优化 蛋白酶 酶活性 

分 类 号：Q786[生物学—分子生物学]