From shake flasks to bioreactors: survival of E. coli cells harboring pGST-hPTH through auto-induction by controlling initial content of yeast extract
文献类型：期刊论文
| 作者 | Jia, Lianghui2; Cheng, Hairong3; Wang, Hengwei3; Luo, Huairong2 ; Yan, Hua1
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| 刊名 | APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
 |
| 出版日期 | 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 |
| 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. |
入库方式： OAI收割
来源：昆明植物研究所
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