From shake flasks to bioreactors: survival of E. coli cells harboring pGST-hPTH through auto-induction by controlling initial content of yeast extract

doi:10.1007/s00253-011-3179-5

	From shake flasks to bioreactors: survival of E. coli cells harboring pGST-hPTH through auto-induction by controlling initial content of yeast extract
	Jia, Lianghui 2; Cheng, Hairong 3; Wang, Hengwei 3; Luo, Huairong2 ; Yan, Hua 1
通讯作者	Yan, H (reprint author), Chinese Acad Sci, Mol Cell Biol Lab, Inst Biochem & Cell Biol, Shanghai Inst Biol Sci, 320 Yueyang Rd, Shanghai 200031, Peoples R China
	2011-05-01
发表期刊	APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
ISSN	0175-7598
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
学科领域	Biotechnology & Applied Microbiology
DOI	10.1007/s00253-011-3179-5
收录类别	SCI
语种	英语
WOS记录号	WOS:000290574700022
引用统计
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
推荐引用方式 GB/T 7714	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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