RRC ID 27480
著者 Isotani K, Kurokawa J, Suzuki F, Nomoto S, Negishi T, Matsuda M, Itoh N.
タイトル Gene cloning and characterization of two NADH-dependent 3-quinuclidinone reductases from Microbacterium luteolum JCM 9174.
ジャーナル Appl Environ Microbiol
Abstract We used the resting-cell reaction to screen approximately 200 microorganisms for biocatalysts which reduce 3-quinuclidinone to optically pure (R)-(-)-3-quinuclidinol. Microbacterium luteolum JCM 9174 was selected as the most suitable organism. The genes encoding the protein products that reduced 3-quinuclidinone were isolated from M. luteolum JCM 9174. The bacC gene, which consists of 768 nucleotides corresponding to 255 amino acid residues and is a constituent of the bacilysin synthetic gene cluster, was amplified by PCR based on homology to known genes. The qnr gene consisted of 759 nucleotides corresponding to 252 amino acid residues. Both enzymes belong to the short-chain alcohol dehydrogenase/reductase (SDR) family. The genes were expressed in Escherichia coli as proteins which were His tagged at the N terminus, and the recombinant enzymes were purified and characterized. Both enzymes showed narrow substrate specificity and high stereoselectivity for the reduction of 3-quinuclidinone to (R)-(-)-3-quinuclidinol.
巻・号 79(4)
ページ 1378-84
公開日 2013-2-1
DOI 10.1128/AEM.03099-12
PII AEM.03099-12
PMID 23263947
PMC PMC3568593
MeSH Actinomycetales / enzymology* Actinomycetales / genetics Amino Acid Sequence Cloning, Molecular Coenzymes / metabolism* DNA, Bacterial / chemistry DNA, Bacterial / genetics Escherichia coli / genetics Molecular Sequence Data NAD / metabolism* Oxidoreductases / genetics* Oxidoreductases / metabolism* Quinuclidines / metabolism* Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / isolation & purification Sequence Alignment Sequence Analysis, DNA Substrate Specificity
IF 4.016
引用数 10
WOS 分野 BIOTECHNOLOGY & APPLIED MICROBIOLOGY MICROBIOLOGY
リソース情報
一般微生物 JCM 3705 JCM 9171 JCM 9174