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| Expression of Paenibacillus macerans α-cyclodextrin Glycosyltraferase in Bacillus subtilis by Maltose Induction |
| ZHANG Jia-yu1,2, WU Dan1,2, CHEN Sheng1,2, CHEN Jian1,2, 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 The α-cgt gene was obtained from Paenibacillus macerans by PCR, and then it was cloned into Escherichia coli-Bacillus subtilis shuttle vector pGJ103 and transformed into B. subtilis WB600. After cultivation for 48 h with shake flask in 1.5% maltose initial medium, the α-CGTase activity of recombinant B. subtilis was 6.1U/ml. In addition, the experiment optimized the culture conditions of the recombinant B. subtilis strain in shaking flask by means of a single factor synthesis and Box-Behnken design (BBD). The analysis predicted the concentrations of maltose, corn starch and yeast extract were at 15 g/L, 13 g/L, and 20 g/L, respectively. In this condition, the experimental result of 17.6 U/ml could be obtained when the cells were cultured for 36 h in shaking flasks. The CGTase activity reached 20 U/ml at 30 h of culture in a 5 L bioreactor (hydrolysis activity was 1.4×104 IU/ml).
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Received: 19 August 2010
Published: 25 December 2010
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