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.
出处
《化学与生物工程》
CAS
2011年第2期63-67,共5页
Chemistry & Bioengineering
关键词
重组毕赤酵母
猪胰岛素前体
拷贝数
生理特性
recombinant Pichia pastoris
porcine insulin precursor
copy number
physiological characterization
