重组枯草芽胞杆菌不对称还原产d-伪麻黄碱

Asymmetric biosynthesis of d-pseudoephedrine by recombinant Bacillus subtilis

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

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

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.

Key words:Bacillus subtilis, carbonyl reductase, whole-cell biotransformation, d-pseudoephedrine