From shake flasks to bioreactors: survival of E. coli cells harboring pGST-hPTH through auto-induction by controlling initial content of yeast extract
Jia, Lianghui2; Cheng, Hairong3; Wang, Hengwei3; Luo, Huairong2; Yan, Hua1
2011-05-01
发表期刊APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
ISSN0175-7598
卷号90期号:4页码:1419-1428
摘要A high content of yeast extract in complex media can cause auto-induction of phage T7 RNA polymerase and the consequent expression of recombinant protein in Escherichia coli BL21(DE3) during long-term cultivation. Our study demonstrated that the auto-induction of recombinant protein varied in different vectors harboring heterologous genes. Trx, GST, and their fusion proteins such as GST-human parathyroid hormone (hPTH), expressed by pET32a (+), were easily auto-induced by media containing a high content of yeast extract; however, rtPA was not easily auto-induced when using pET22b (+), although both pET systems were under the control of T7lac promoter. Furthermore, the auto-induction of GST-hPTH may start within 1-2 h after inoculation in bioreactors, which is a deficiency in the scale-up from shake flasks to bioreactors. Our results indicated that too much yeast extract in bioreactor cultivations may be responsible for the early auto-induction of target proteins and consequent loss of cell viability and plasmid instability. To achieve a satisfactory yield, host cells with both high cell viability and plasmid stability were necessary for the starter cultures in shake flasks and pre-induction cultures in bioreactors. This could be achieved simply by controlling the initial content of yeast extract and its subsequent supplementation.
关键词Auto-induction E. Coli Pet Systems Yeast Extract Cell Viability Plasmid Stability
资助信息SA-SIBS; Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences[2009KIP501]
学科领域Biotechnology & Applied Microbiology
DOI10.1007/s00253-011-3179-5
收录类别SCI
语种英语
WOS研究方向Biotechnology & Applied Microbiology
WOS类目Biotechnology & Applied Microbiology
WOS记录号WOS:000290574700022
引用统计
被引频次:5[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.kib.ac.cn/handle/151853/5343
专题植物化学与西部植物资源持续利用国家重点实验室
作者单位1.Chinese Acad Sci, Mol Cell Biol Lab, Inst Biochem & Cell Biol, Shanghai Inst Biol Sci, Shanghai 200031, Peoples R China
2.Chinese Acad Sci, State Key Lab Phytochem & Plant Resources W China, Ctr Drug Screening & Res, Kunming Inst Bot, Kunming 650204, Yunnan, Peoples R China
3.Shanghai Jiao Tong Univ, Key Lab Microbial Metab, Minist Educ, Sch Life Sci, Shanghai 200240, Peoples R China
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Jia, Lianghui,Cheng, Hairong,Wang, Hengwei,et al. From shake flasks to bioreactors: survival of E. coli cells harboring pGST-hPTH through auto-induction by controlling initial content of yeast extract[J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,2011,90(4):1419-1428.
APA Jia, Lianghui,Cheng, Hairong,Wang, Hengwei,Luo, Huairong,&Yan, Hua.(2011).From shake flasks to bioreactors: survival of E. coli cells harboring pGST-hPTH through auto-induction by controlling initial content of yeast extract.APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,90(4),1419-1428.
MLA Jia, Lianghui,et al."From shake flasks to bioreactors: survival of E. coli cells harboring pGST-hPTH through auto-induction by controlling initial content of yeast extract".APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 90.4(2011):1419-1428.
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