High cell-density fermentation of shark hepatical stimulator analogue in Escherichia coli.
- Author:
Boping YE
1
;
Zheng PAN
;
Huaibiao LI
;
Ying WANG
;
Heng ZHENG
;
Wutong WU
Author Information
1. School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China. yebp2001@yahoo.com.cn
- Publication Type:Journal Article
- MeSH:
Animals;
Cloning, Molecular;
Escherichia coli;
genetics;
metabolism;
Fermentation;
Liver;
chemistry;
Peptides;
genetics;
metabolism;
Recombinant Proteins;
biosynthesis;
genetics;
Sharks;
metabolism
- From:
Chinese Journal of Biotechnology
2009;25(9):1371-1378
- CountryChina
- Language:Chinese
-
Abstract:
The potential effects of recombinant shark hepatical stimulator analogue (r-sHSA) in liver disease have been revealed in our previous studies. In order to further evaluate its clinic application, we carried out high cell-density fermentation in 5 L fermentor to get enough products. Based on the trials in shaking flask, we optimized the parameters for 5 L fermentor, including medium composition, medium supplement, inducer concentration and induction time, etc. In detail, the improved LB medium (0.97% glycerol, 0.91% yeast extract, 0.72% tryptone, 0.782% KH2PO4, 0.267% K2HPO4.3H2O, 0.062% MgSO4.7H2O, 0.5% NaCl, pH 7.0) is chosen to cultivate the engineering bacteria with the constant fermentation condition (pH 7.0, and the dissolved oxygen concentration is about 25%-30%). When bacterial culture reaches exponential phase, the modified feeding medium (620 g/L glycerol, 94.8 g/L tryptone, 3.3 mL/L trace elements, and 7.5 g/L MgSO4.7H2O) is then supplied through the method of exponential fed-batch mode. After the optical density (OD600) of engineering bacterial culture reaches to 23, the ultimately concentration of 0.5 mmol/L IPTG is added to induce the expression of r-sHSA for 6 h. Results show that the amount of r-sHSA production is (2.662 +/- 0.041) g/L, which is about 13.7 folds of the one optimized before.
