【摘要】 目的：建立一种在150L发酵罐中提高rhIL-11蛋白产出率的控制方法。方法：研究在毕赤酵母表达rhIL-11的发酵过程中,分别采用间歇法（甲醇浓度0．5%,每24h诱导一次）、分批补料法（甲醇补料速度在8h内从2．88ml/min提高至8．64ml/min）以及恒甲醇浓度法（利用甲醇浓度传感器检测并控制甲醇补料速度使甲醇浓度始终保持0．5%左右）的不同甲醇诱导模式,对工程菌生长及目的蛋白表达的影响。结果：采用间歇法的菌浓OD₆₀₀可达到80、rhIL-11表达浓度可达到0．1mg/ml；分批补料法的菌浓OD₆₀₀可达到240、rhIL-11表达浓度可达到1．0mg/ml；恒甲醇浓度法的菌浓OD₆₀₀可达到300、rkIL-11表达浓度可达到1．0mg/ ml。结论：rhIL-11蛋白产出率,恒甲醇浓度法（约0．12g/d·L）比间歇法（约0．01g/d·L）和分批补料法（约0．09g/d·L）都高,分别提高了12～13倍和30～40%。更多还原

【Abstract】 Objective:To obtain a method to increased the yield of rhIL-11 in a 150 L fementor.Methods:Study the influence of different metha- nol feeding modes on the fermentation of rhIL-11 which expressed by Pichia pastoris,such as intermittent mode（methanol concentratian and fre- quency of methanol induction were 0.5% and in every 24h,respectively）,fed-batch cultivation mode（methanol feeding rate were improved 2.88ml/min to 8.64ml/min in 8h）and constant-methanol concentration mode（using a methanol sensor to monitor and control methanol feeding rate to maintain a 0.5% methanol concentration）,on the growth of P/ch/a pastor/s and expression of target protein.Results:A density(OD₆₀₀) and a expression level of intermittent mode were about 80 and 0.lmg/ml,respectively.A density(OD₆₀₀)and a expression level of fed-batch cultivation mode were about 240 and 1.0rag/nil,respectively.A density(OD₆₀₀)and a expression level of constant-methanol concentration mode were about 300 and 1.0mg/ml,respectively.Conclusions:The rhIL-11 productivity level were improved by constant-methanol concentration mode（about 0.12g/d·L）compared with intermittent（about 0.01 g/d·L）and fed-batch cultivation modes（about 0.09g/d·L）.It was showed that the rhIL-11 productivity level was improved up to 12-13 times and 30-40% compared with intermittent and fed-batch cultivation methanol feeding respectively.更多还原