BioChem - DOS

Gene cloning and characterization of two NADH-dependent 3-quinuclidinone reductases from Microbacterium luteolum JCM 9174

Applied and environmental microbiology

Isotani, Kentaro; Kurokawa, Junji; Suzuki, Fumiko; Nomoto, Syunsuke; Negishi, Takashi; Matsuda, Michiko; Itoh, Nobuya

<P><B>ABSTRACT</B><P> We used the resting-cell reaction to screen approximately 200 microorganisms for biocatalysts which reduce 3-quinuclidinone to optically pure ( <I>R</I> )-(&#x2212;)-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 <I>bacC</I> 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 <I>qnr</I> 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 ( <I>R</I> )-(&#x2212;)-3-quinuclidinol. </P></P>

2013

American Society for Microbiology

0099-2240

1098-5336

79

4

pp.1378-1384

10.1128/aem.03099-12

http://click.ndsl.kr/servlet/OpenAPIDetailView?keyValue=05787966&target=NART&cn=NART65576286

https://aem.asm.org/content/aem/79/4/1378.full.pdf