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题名: From shake flasks to bioreactors: survival of E. coli cells harboring pGST-hPTH through auto-induction by controlling initial content of yeast extract
作者: Jia, Lianghui2; Cheng, Hairong3; Wang, Hengwei3; null(罗怀容)2; Yan, Hua1
刊名: APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
关键词: Auto-induction ; E. coli ; pET systems ; Yeast extract ; Cell viability ; Plasmid stability
英文摘要: 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.
出版日期: 2011-05-01
卷号: 90, 期号:4, 页码:1419-1428
语种: 英语
收录类别: SCI
学科分类: Biotechnology & Applied Microbiology
ISSN号: 0175-7598
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.kib.ac.cn/handle/151853/5343
Appears in Collections:植物化学与西部植物资源持续利用国家重点实验室_期刊论文

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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

Recommended Citation:
Jia, LH; Cheng, HR; Wang, HW; Luo, HR; Yan, H.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,2011,90(4):1419-1428
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