人三叶因子3在毕赤酵母中表达条件的研究  

Studies on the Expression Condition of Human Trefoil Factor 3 in Pichia pastoris

作　　者：王艳茹[1] 方敏[1] 安琳[1] 李令媛[1] 茹炳根[1] 

机构地区：[1]北京大学生命科学学院蛋白质工程国家重点实验室,北京100871

出　　处：《生物工程学报》2001年第6期648-651,共4页Chinese Journal of Biotechnology

摘　　要：为提高人三叶因子 3 (HumanTrefoilfactor 3 ,hTFF3 )在毕赤酵母中的表达量 ,研究了转化子生长的培养条件 ,包括不同碳源对转化子生长的影响和接种量、甲醇浓度、pH值、摇瓶转速及不同诱导时间对人三叶因子 3表达的影响。结果表明转化子在生长阶段加入葡萄糖生长旺盛 ,培养 14h后OD6 0 0 就可达到 5 0。在 10 0mL生长培养基上的菌液以 1∶1接入诱导培养基时蛋白表达量最高 ;转化子在 1%的甲醇、pH6 0、摇瓶转速 2 4 0r min的条件下诱导4 8h ,菌体密度OD6 0 0 为 15 ,目的蛋白表达量达到 2 0mg L。用 5L发酵罐进行了高密度发酵 ,经 2 %甲醇 3 2h诱导 ,最终菌体密度OD6 0 0 达到 12 0 ,每升发酵液中含目的蛋白 10 0mg。In order to enhance the expression level of human trefoil factor 3 (hTFF3) in %Pichia pastoris,%we optimized the transformant growth conditions in shake flasks including carbon sources in growth medium,inoculation ratio,methanol conentration,pH rotation speed and inducing time.The transformant could grow on the glucose to %OD% 600 5 0 after 14 hours inoculation.The best inoculation ration of 100mL growth medium to the induction medium was 1∶1.The expression level of dimeric human trefoil factor 3 induction with 1% methanol for 48 hours at pH 6 0,agitation speed 240r/min could reach 20mg/L with OD 600 15.The protein was expressed in 5-liter fermentor with 2% methanol induction for 32 hours,finally the cell density reached %OD% 600 120 100mg/L of recombinant hTFF3 was obtained in the supernatant.

关 键 词：三叶因子3 毕赤酵母 表达 发酵 培养条件 

分 类 号：Q986[生物学—人类学]