RRC ID 5738
著者 Uzura A, Nomoto F, Sakoda A, Nishimoto Y, Kataoka M, Shimizu S.
タイトル Stereoselective synthesis of (R)-3-quinuclidinol through asymmetric reduction of 3-quinuclidinone with 3-quinuclidinone reductase of Rhodotorula rubra.
ジャーナル Appl Microbiol Biotechnol
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.
巻・号 83(4)
ページ 617-26
公開日 2009-6-1
DOI 10.1007/s00253-009-1902-2
PMID 19234697
MeSH Alcohol Oxidoreductases / genetics Alcohol Oxidoreductases / isolation & purification Alcohol Oxidoreductases / metabolism* Amino Acid Sequence Base Sequence Biotransformation Cloning, Molecular DNA, Fungal / chemistry DNA, Fungal / genetics Escherichia coli / genetics Fungal Proteins / genetics Fungal Proteins / isolation & purification Fungal Proteins / metabolism* Gene Expression Glucose 1-Dehydrogenase / genetics Glucose 1-Dehydrogenase / metabolism Molecular Sequence Data Quinuclidines / metabolism* Rhodotorula / enzymology* Rhodotorula / genetics Sequence Analysis, DNA Sequence Homology, Amino Acid Stereoisomerism
IF 3.53
引用数 26
WOS 分野 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
リソース情報
一般微生物 JCM3782