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).
abstract: 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.
authors.affiliation: Department of Biotechnology, Faculty of Engineering, Biotechnology Research Center, Toyama Prefectural University, 5180 Kurokawa, Imizu, Toyama 939-0398, Japan. nbito@pu-toyama.ac.jp.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Isotani
authors.forename: Junji
authors.initials: J
authors.lastname: Kurokawa
authors.forename: Nobuya
authors.initials: N
authors.lastname: Itoh
mesh_terms: D000192:Actinomycetales / Q000201:enzymology*; D000426:Alcohol Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D055162:Biocatalysis; D004926:Escherichia coli / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D009247:NADH, NADPH Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D009904:Optical Rotation; D011812:Quinuclidines / Q000737:chemistry / Q000378:metabolism*; D013237:Stereoisomerism; D013997:Time Factors
chemical_list: D011812:Quinuclidines; C474540:3-quinuclidinol; D000426:Alcohol Dehydrogenase; D009247:NADH, NADPH Oxidoreductases