文献类型：期刊文献

中文题名：毕赤酵母高密度表达重组猪胰岛素前体的研究

英文题名：High-density Expression of Recombinant Porcine Insulin Precursor by Pichia pastoris

作者：刘玉伟[1];黄明志[1];庄英萍[1];储炬[1];张嗣良[1]

机构：[1]华东理工大学生物反应器工程国家重点实验室国家生化工程技术研究中心(上海),上海200237

年份：2007

卷号：34

期号：1

起止页码：75

中文期刊名：微生物学通报

外文期刊名：Microbiology China

收录：CSTPCD;;北大核心:【北大核心2004】；CSCD:【CSCD2011_2012】；

语种：中文

中文关键词：巴斯德毕赤酵母;高密度;重组猪胰岛素前体

外文关键词：Pichia pastoris, High-density fermentation, Recombinant porcine insulin precursor

摘要：对摇瓶和50L罐上的重组菌毕赤酵母(Pichia pastoris)表达猪胰岛素前体(PIP)的发酵过程进行了研究。摇瓶发酵中,最佳诱导周期为60 h左右,诱导期甲醇的最佳加入量为每日2．0%～2．5%。50L发酵过程分为批发酵、补料和诱导表达3个阶段。生长期(批发酵和补料阶段)细胞干重与培养时间的关系可用模型y= 0．6525e^(0．1909t)来描述。在批发酵阶段和补料阶段,流加的氨水和甘油几乎全部用来合成菌体和维持,没有其他副产物产生。诱导表达阶段流加的氨水和甲醇分别约有80%和70%被菌体利用。将摇瓶与发酵罐的实验结果进行了比较,发现摇瓶发酵的限制因子很可能是溶氧,而罐发酵的限制因子为碳源,因此,将摇瓶实验的结果放大到发酵罐时调整了控制策略,加大了甲醇的补料速率,最终PIP浓度达到1．72g/L。
The recombinant porcine insulin precursor （PIP） produced by Pichia pastoris in shake-flask and 50L fermenter was investigated respectively. The results indicated that 60h induction time length and 2.0% -2.5% methanol addition every day was optimum in shake-flask. The process in 50L fermenter was consisted of batch, feed-batch and induction phases. The relationship between dry cell weight （y） and culture time （t） in growth phase （batch and feed-batch phase） could be described by model y = 0. 6525e^0.1907t. Glycerol and ammonia were almost used for cell growth and maintain, and no by-preduct was observed in batch and fed-batch phase. Only 80% ammonia and 70% methanol were used by cell in induction phase. By comparison the results of shake-flask and 50L fermenter, it was concluded that the limiting factor in the fermentation of shake-flask and 50L fermenter was dissolved oxygen （DO） and. carbon source, respectively. When scaling the result of shake-flask to 50L fermenter, the control strategy was adapted for 50L fermenter by increasing the feed rate of methanol and the maximum PIP concentration reached 1.72 g/L.