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Int. J. Mol. Sci. 2012, 13(10), 13542-13553; doi:10.3390/ijms131013542
Article

Production of (R)-3-Quinuclidinol by E. coli Biocatalysts Possessing NADH-Dependent 3-Quinuclidinone Reductase (QNR or bacC) from Microbacterium luteolum and Leifsonia Alcohol Dehydrogenase (LSADH)

,
 and
*
Department of Biotechnology, Faculty of Engineering, Biotechnology Research Center, Toyama Prefectural University, 5180 Kurokawa, Imizu, Toyama 939-0398, Japan
* Author to whom correspondence should be addressed.
Received: 30 August 2012 / Revised: 27 September 2012 / Accepted: 11 October 2012 / Published: 19 October 2012
(This article belongs to the Special Issue Organic Synthesis Using Biocatalyst)
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Abstract

We found two NADH-dependent reductases (QNR and bacC) in Microbacterium luteolum JCM 9174 (M. luteolum JCM 9174) that can reduce 3-quinuclidinone to optically pure (R)-(−)-3-quinuclidinol. Alcohol dehydrogenase from Leifsonia sp. (LSADH) was combined with these reductases to regenerate NAD+ to NADH in situ in the presence of 2-propanol as a hydrogen donor. The reductase and LSADH genes were efficiently expressed in E. coli cells. A number of constructed E. coli biocatalysts (intact or immobilized) were applied to the resting cell reaction and optimized. Under the optimized conditions, (R)-(−)-3-quinuclidinol was synthesized from 3-quinuclidinone (15% w/v, 939 mM) giving a conversion yield of 100% for immobilized QNR. The optical purity of the (R)-(−)-3-quinuclidinol produced by the enzymatic reactions was >99.9%. Thus, E. coli biocatalysis should be useful for the practical production of the pharmaceutically important intermediate, (R)-(−)-3-quinuclidinol.
Keywords: 3-quinuclidinone reductase (QNR and bacC); Microbacterium luteolum JCM 9174; Leifsonia alcohol dehydrogenase (LSADH); (R)-(−)-3-quinuclidinol; optically pure alcohol; immobilized biocatalyst 3-quinuclidinone reductase (QNR and bacC); Microbacterium luteolum JCM 9174; Leifsonia alcohol dehydrogenase (LSADH); (R)-(−)-3-quinuclidinol; optically pure alcohol; immobilized biocatalyst
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).
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Isotani, K.; Kurokawa, J.; Itoh, N. Production of (R)-3-Quinuclidinol by E. coli Biocatalysts Possessing NADH-Dependent 3-Quinuclidinone Reductase (QNR or bacC) from Microbacterium luteolum and Leifsonia Alcohol Dehydrogenase (LSADH). Int. J. Mol. Sci. 2012, 13, 13542-13553.

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