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- Title
Stereoselective synthesis of ( R)-3-quinuclidinol through asymmetric reduction of 3-quinuclidinone with 3-quinuclidinone reductase of Rhodotorula rubra.
- Authors
Uzura, Atsuko; Nomoto, Fumiki; Sakoda, Akiko; Nishimoto, Yukifumi; Kataoka, Michihiko; Shimizu, Sakayu
- Abstract
A novel nicotinamide adenine dinucleotide phosphate-dependent carbonyl reductase, 3-quinuclidinone reductase, was isolated from Rhodotorula rubra JCM3782. The enzyme catalyzes the asymmetric reduction of 3-quinuclidinone to ( R)-3-quinuclidinol. The gene encoding the enzyme was also cloned and sequenced. A 819-bp nucleotide fragment was confirmed to be the gene encoding the 3-quinuclidinone reductase by agreement of the internal amino acid sequences of the purified enzyme. The gene encodes a total of 272 amino acid residues, and the deduced amino acid sequence shows similarity to those of several short-chain dehydrogenase/reductase family proteins. An expression vector, pWKLQ, which contains the full length 3-quinuclidinone reductase gene was constructed. Using Escherichia coli cells coexpressing the 3-quinuclidinone reductase and glucose dehydrogenase (cofactor regeneration enzyme) genes, 618 mM 3-quinuclidinone was almost stiochiometrically converted to ( R)-3-quinuclidinol with an >99.9% enantiomeric excess within 21 h of reaction.
- Subjects
CARBONYL reductase; RHODOTORULA rubra; CATALYSIS; ENZYMES; AMINO acid sequence; GENETIC vectors; ESCHERICHIA coli; DEHYDROGENASES
- Publication
Applied Microbiology & Biotechnology, 2009, Vol 83, Issue 4, p617
- ISSN
0175-7598
- Publication type
Article
- DOI
10.1007/s00253-009-1902-2