| 重组枯草芽胞杆菌不对称还原产d-伪麻黄碱 |
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| 引用本文： | 彭艳红,张梁,丁重阳,王正祥,石贵阳.重组枯草芽胞杆菌不对称还原产d-伪麻黄碱[J].生物工程学报,2011,27(7):1082-1091. |
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| 作者姓名： | 彭艳红 张梁 丁重阳 王正祥 石贵阳 |
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| 作者单位： | 江南大学工业生物技术教育部重点实验室，无锡，214122;江南大学生物工程学院生物资源与生物能源研究中心，无锡，214122 |
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| 基金项目： | 国家自然科学基金 (No. 30770054)，2008年度江苏省高校“青蓝工程”科技创新团队资助。 |
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| 摘 要： | 为了实现羰基还原酶基因mldh在枯草芽胞杆菌Bacillus subtilis中的表达并通过细胞内的葡萄糖脱氢酶完成辅酶的再生，以枯草芽胞杆菌rpsD基因的启动子PrpsD和终止子TrpsD为表达元件，将羰基还原酶基因mldh连接至构建好的质粒(pHY300plk-PrpsD-TrpsD上，得到质粒pHY300plk-PrpsD-mldh-TrpsD；进一步将重组质粒转化入B. subtilis Wb600中获得重组菌B. subtilis Wb600 (pHY300plk-PrpsD-mldh-Trps
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| 关 键 词： | 枯草芽胞杆菌，羰基还原酶，全细胞生物转化，d-伪麻黄碱 |
| 收稿时间： | 2010/10/8 0:00:00 |
Asymmetric biosynthesis of d-pseudoephedrine by recombinant Bacillus subtilis |
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| Yanhong Peng,Liang Zhang,Zhongyang Ding,Zhengxiang Wang and Guiyang Shi.Asymmetric biosynthesis of d-pseudoephedrine by recombinant Bacillus subtilis[J].Chinese Journal of Biotechnology,2011,27(7):1082-1091. |
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| Authors: | Yanhong Peng Liang Zhang Zhongyang Ding Zhengxiang Wang and Guiyang Shi |
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| Institution: | Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China; Center for Bioresource & Bioenergy, School of Biotechnology, Jiangnan University, Wuxi 214122, China;Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China; Center for Bioresource & Bioenergy, School of Biotechnology, Jiangnan University, Wuxi 214122, China;Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China; Center for Bioresource & Bioenergy, School of Biotechnology, Jiangnan University, Wuxi 214122, China;Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China; Center for Bioresource & Bioenergy, School of Biotechnology, Jiangnan University, Wuxi 214122, China;Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China; Center for Bioresource & Bioenergy, School of Biotechnology, Jiangnan University, Wuxi 214122, China |
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| Abstract: | In order to successfully express the carbonyl reductase gene mldh in Bacillus subtilis and complete coenzyme regeneration by B. subtilis glucose dehydrogenase, the promoter PrpsD and the terminator TrpsD from B. subtilis rpsD gene were used as the expression cassette to be a recombinant plasmid pHY300plk-PrpsD-TrpsD. After that, the carbonyl reductase gene mldh was inserted into the previous plasmid and a plasmid pHY300plk-PrpsD-mldh-TrpsD was achieved, followed by transformed into B. subtilis Wb600 to obtain a recombinant B. subtilis Wb600 (pHY300plk-PrpsD-mldh-TrpsD). Subsequently, the results for whole-cell biotransformation from recombinant B. subtilis showed that it could be used to catalyze MAK (1-phenyl-1-keto-2-methylaminopropane) to d-pseudoephedrine in the presence of glucose. The yield of d-pseudoephedrine could be up to 97.5 mg/L and the conversion rate of MAK was 24.1%. This study indicates the possibility of biotransformation production of d-pseudoephedrine from recombinant B. subtilis. |
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| Keywords: | Bacillus subtilis carbonyl reductase whole-cell biotransformation d-pseudoephedrine |
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