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 |