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

	From shake flasks to bioreactors: survival of E. coli cells harboring pGST-hPTH through auto-induction by controlling initial content of yeast extract
	Jia, LH; Cheng, HR; Wang, HW; Luo, HR; Yan, H
刊名	APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
	2011
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
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.,yanhua@sibs.ac.cn
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
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
语种	英语
WOS记录号	WOS:000290574700022
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/773]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Jia, LH,Cheng, HR,Wang, HW,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, LH,Cheng, HR,Wang, HW,Luo, HR,&Yan, H.(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, LH,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.