[Asymmetric biosynthesis of d-pseudoephedrine by recombinant Bacillus subtilis].
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 Laboratory of Industrial Biotechnology, Ministry ofEducation, Jiangnan University, Wuxi 214122, China
Yanhong
Y
Peng
Liang
L
Zhang
Zhongyang
Z
Ding
Zhengxiang
Z
Wang
Guiyang
G
Shi
D000429:Alcohol Oxidoreductases / Q000235:genetics; D001412:Bacillus subtilis / Q000235:genetics* / Q000378:metabolism*; D042843:Glucose 1-Dehydrogenase / Q000737:chemistry / Q000378:metabolism; D016254:Mutagenesis, Insertional; D054199:Pseudoephedrine / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D011995:Recombination, Genetic
D011994:Recombinant Proteins; D054199:Pseudoephedrine; D000429:Alcohol Oxidoreductases; D042843:Glucose 1-Dehydrogenase