拷贝数对毕赤酵母重组菌表达猪胰岛素前体的影响  

Effects of Copy Number on Expression of Porcine Insulin Precursor (PIP) by Recombinant Pichia Pastoris

作　　者：陈丽[1] 郭美锦[1] 储炬[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国家生化工程技术研究中心,上海200237

出　　处：《化学与生物工程》2011年第2期63-67,共5页Chemistry & Bioengineering

摘　　要：通过5 L发酵罐补料分批培养,分析了猪胰岛素前体(PIP)基因拷贝数对毕赤酵母生长、细胞存活率和基因稳定性的影响。结果表明,在甘油阶段和甲醇诱导初期拷贝数对毕赤酵母的生长无显著影响,但在甲醇诱导24 h后高拷贝数毕赤酵母菌株(12拷贝重组菌A2和18拷贝重组菌A3)的比生长速率明显比低拷贝数菌株(1拷贝重组菌G1和6拷贝重组菌G6)的比生长速率慢,A3菌株细胞存活率仅为78%,且其外源基因丢失率高达19.4%;4种拷贝数毕赤酵母重组菌中,12拷贝重组菌A2的PIP蛋白产量最高,为702 mg.L-1。The effects of porcine insulin precursor（PIP） gene copy number on the growth,cell viability and genetic stability of multi-copy Pichia pastoris cells were studied in 5 L fermenter fed-batch culture.The results showed that there were no significant changes in cell growth of multi-copy Pichia pastoris from the glycerol phase to the early methanol induction phase.However,after 24 h of methanol induction,the specific growth rates of both recombinant strain A2 harboring 12-copy PIP gene and strain A3 harboring 18-copy PIP gene remarkably decreased as compared with the low multi-copy strains G1（1-copy PIP gene） and G6（6-copy PIP gene）.Also,the cell viability of the PIP gene strain A3 was only 78% and its PIP gene loss rate was up to 19.4% in genetic stability experiments.The maximum PIP production of 702 mg·L-1 was achieved by the strain A2 grown in fed-batch culture among four multiple PIP copies Pichia pastoris strains tested here.

关 键 词：重组毕赤酵母 猪胰岛素前体 拷贝数 生理特性 

分 类 号：TQ920.6[轻工技术与工程—发酵工程]