Reference - Detail
|Author||Isotani K, Kurokawa J, Itoh N.|
|Title||Production of (R)-3-quinuclidinol by E. coli biocatalysts possessing NADH-dependent 3-quinuclidinone reductase (QNR or bacC) from Microbacterium luteolum and Leifsonia alcohol dehydrogenase (LSADH).|
|Journal||Int J Mol Sci|
We found two NADH-dependent reductases (QNR and bacC) in Microbacterium luteolum JCM 9174 (M. luteolum JCM 9174) that can reduce 3-quinuclidinone to optically pure (R)-(-)-3-quinuclidinol. Alcohol dehydrogenase from Leifsonia sp. (LSADH) was combined with these reductases to regenerate NAD+ to NADH in situ in the presence of 2-propanol as a hydrogen donor. The reductase and LSADH genes were efficiently expressed in E. coli cells. A number of constructed E. coli biocatalysts (intact or immobilized) were applied to the resting cell reaction and optimized. Under the optimized conditions, (R)-(-)-3-quinuclidinol was synthesized from 3-quinuclidinone (15% w/v, 939 mM) giving a conversion yield of 100% for immobilized QNR. The optical purity of the (R)-(-)-3-quinuclidinol produced by the enzymatic reactions was >99.9%. Thus, E. coli biocatalysis should be useful for the practical production of the pharmaceutically important intermediate, (R)-(-)-3-quinuclidinol.
|MeSH||Actinomycetales / enzymology* Alcohol Dehydrogenase / genetics Alcohol Dehydrogenase / metabolism* Biocatalysis Escherichia coli / metabolism* Genetic Vectors / genetics Genetic Vectors / metabolism NADH, NADPH Oxidoreductases / genetics NADH, NADPH Oxidoreductases / metabolism* Optical Rotation Quinuclidines / chemistry Quinuclidines / metabolism* Stereoisomerism Time Factors|
|WOS Category||CHEMISTRY, MULTIDISCIPLINARY BIOCHEMISTRY & MOLECULAR BIOLOGY|
|General Microbes||JCM 9174|