培养条件对重组毕赤酵母高密度表达猪胰岛素前体的影响  

Culture Conditions Affecting High Level Expression of Porcine Insulin Precursor(PIP) by Recombinant Pichia pastoris in High Cell Density Fermentation

作　　者：姚钰舜[1] 储炬[1] 杭海峰[1] 庄英萍[1] 张嗣良[1] 

机构地区：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

出　　处：《华东理工大学学报（自然科学版）》2006年第4期397-401,共5页Journal of East China University of Science and Technology

基　　金：国家高技术研究发展计划(863计划)(2002AA217021);上海市重点学科资助项目

摘　　要：研究了基因工程菌P ich ia p astotis高密度、高表达的培养条件。在全合成摇瓶培养基的基础上,考察了诱导过程中甲醇浓度、温度、pH、(NH4)2SO4及磷酸盐浓度等因素对猪胰岛素前体(P IP)表达的影响。研究表明:甲醇的最佳诱导浓度为6 g/L,过高或过低的甲醇浓度都不利于菌体生长和蛋白表达;在菌体生长和蛋白表达阶段,pH应分别控制在5.5和4.0;诱导阶段,培养温度下降到28°C,可以提高重组蛋白质的表达量。上述结果用于15 L全自动发酵罐实验,P IP表达水平达到了1.69 g/L,是摇瓶结果的17倍。The cultivation conditions for porcine insulin precursor （PIP） production by recombinant Pichia pastoris were optimized in shake flasks with synthetic fermentation media. The influences of pH, temperature, the concentrations of methanol, （NH4）2SO4, and phosphate on the expression of PIP and the cell growth were investigated. The optimum concentration of methanol is 6 g/L. The optimal pH value is 5.5 at cell growth phase and 4. 0 in expression phase. Meanwhile, reducing the culture temperature to 28 ℃ in induction phase can increase the PIP expression level. The above results are successfully applied to a 15 L fermenter, where the PIP concentration reach 1.69 g/L, which is 17 times as high as that in the shake flasks.

关 键 词：毕赤酵母 猪胰岛素前体 高密度培养 高表达 

分 类 号：Q939.9[生物学—微生物学]