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

CORC > 昆明植物研究所 > 中国科学院昆明植物研究所 > 植物化学与西部植物资源持续利用国家重点实验室

	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
刊名	APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
	2011-05-01
卷号	90 期号:4 页码:1419-1428
关键词	Auto-induction E. coli pET systems Yeast extract Cell viability Plasmid stability
ISSN号	0175-7598
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
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.
学科主题	Biotechnology & Applied Microbiology
类目[WOS]	Biotechnology & Applied Microbiology
研究领域[WOS]	Biotechnology & Applied Microbiology
关键词[WOS]	3-(4,5-DIMETHYLTHIAZOL-2-YL)-2,5-DIPHENYLTETRAZOLIUM BROMIDE MTT ; RECOMBINANT ESCHERICHIA-COLI ; ENHANCE PLASMID STABILITY ; TRANSLATION INITIATION ; CELLULAR REDUCTION ; PROTEIN EXPRESSION ; COPY NUMBER ; CODON BIAS ; GLUCOSE ; RNA
收录类别	SCI
资助信息	SA-SIBS; Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences[2009KIP501]
语种	英语
WOS记录号	WOS:000290574700022
公开日期	2012-04-10
内容类型	期刊论文
源URL	[http://ir.kib.ac.cn/handle/151853/5343]
专题	昆明植物研究所_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.