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| Optimization of Fermentation and Induction Conditions of Recombinant E.coli BL21(DE3)/pET30a(+)hrpNEcc |
| ZHANG Shu1,2,WANG Min2,HAN Mei-lin2,MA Rong-cai2,CHEN Qiang1,GAO Jun-lian2 |
1.Department of Microbiology, College of Resources and Environment Science, Sichuan Agricultural University, Ya′an 625014,China
2.Agro-Biotechnology Research Centre, Beijing Academy of Agriculture and Forestry, Beijing 100097,China |
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Abstract In order to obtain high yield of the HrpNEcc protein with a lower total cost, fermentation and lactose induction conditions for recombinant E. coli BL21(DE3)/pET30a(+)hrpN Ecc were optimized in flasks and the recombinant E.coli was fermentated in 7L fermenter. The optimized incoulum concentration was 5% and the optimized nutrient medium was TB medium. The HrpNEcc protein yield reached 417.60mg/L by adding 3g/L exogenous inducer lactose in the growth prophase of log-phase for the recombinant E.coli. The HrpNEcc protein yield was higher 36.73% than that of the case of no any inducer, and was higher 16.85% than that of the case of adding IPTG. The wet weight of cell pellet of the recombinant E.coli reached 57.24g/L after fermentation in 7L fermenter, the HrpNEcc protein reached 3.29g/L, about 50.2% of total cellular protein.
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Received: 25 May 2009
Published: 29 October 2009
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Corresponding Authors:
GAO Jun-Lian
E-mail: gaojunlian@baafs.net.cn;qiangchencq@yahoo.com.cn
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