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| Cloning,Expression and Fermentation Optimization of Thermobifida fusca Trehalose Synthase Gene in E.coli |
| LUO Feng1,2, DUAN Xu-guo1,2, SU Ling-qia1,2, WU Jing1,2 |
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 214122, China |
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Abstract In the present study, the trehalose synthase gene of Thermobifida fusca was cloned and expressed in Escherichia coli BL21(DE3). The optimized composition of fermentation medium and culture conditions for the recombinant E. coli were investigated in shake flasks. The optimal fermentation medium was as follows: 12 g/L glycerol, 24 g/L yeast extract, 12 g/L peptone, 60 mmol/L PO43-, 2 mmol/L Zn2+. And the optimal culture conditions were that: volume 20 ml in 250 ml shake flask, induction temperature 25℃, induced by 0.4 mmol/L IPTG at 2 h of culture. Under these conditions, the maximal enzyme activity reached 28.6 U/ml, which was 3.4 times as high as that not optimized.
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Received: 02 May 2013
Published: 25 August 2013
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