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中国生物工程杂志

China Biotechnology
China Biotechnology  2011, Vol. 31 Issue (9): 55-61    DOI:
    
Fermentation Optimization on Flask-scale and Secretional Expression of Recombinant Cutinase-CBM in E.coli
GUO Sen1,2, WU Dan1,2, CHEN Sheng1,2, WU Jing1,2, CHEN Jian1,2
1. State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China;
2. School of Biotechnology and Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 2141221, China
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Abstract  

On the basis of TB fermentation medium, an optimized fermentation strategy of cutinase-CBM by E.coli was developed through single factor analyzing and orthogonal design, which can be illustrated as follows: glycerol 5g/L, peptone 16 g/L, MgSO4·7H2O 2.5 mmol/L, K2HPO4 13.7 g/L, KH2PO4 1.53 g/L, 1 g/L lactose and 0.75 g/L glycine (final concentration) were added in the prometaphase of logarithmic phase of growth curve, and the fermentation period lasted 48h under 30℃. An extracellular enzyme activity of 63 U/ml was reached, about 3 times that of the control (20 U/ml). Further efforts were made to investigate the influence of other factors, such as heat-shock, osmotic agent, and temperature, on the secretional expression of cutinase-CBM. It is found that with the addition of 75 mmol/L L-proline, heat shocked for 1h at 37℃ or 0.5h at 47℃, then shifted to 25℃, the final extracellular production of cutinase-CBM can reach 90U/ml, 4 times that of the control cultivated at constant temperature.



Key wordsCutinase-CBM      Orthogonal design      Heat-shock      Osmotic agent      Temperature-shifted fermentation     
Received: 26 May 2011      Published: 25 September 2011
ZTFLH:  Q786  
Cite this article:

GUO Sen, WU Dan, CHEN Sheng, WU Jing, CHEN Jian. Fermentation Optimization on Flask-scale and Secretional Expression of Recombinant Cutinase-CBM in E.coli. China Biotechnology, 2011, 31(9): 55-61.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2011/V31/I9/55


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