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.
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Acknowledgments
This work was supported in part by a grant-in-aid for Developmental Research from the New Energy and Industrial Technology Development Organization (NEDO). This work was also supported in part by a grant-in-aid for Scientific Research, no. 20380051 (to MK), from the Japan Society for the Promotion of Science (JSPS), and by the Targeted Proteins Research Program (TPRP) of the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan. The technical assistance of Ms. N. Kitamura and Mr. H. Saito of this laboratory is greatly appreciated.
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Uzura, A., Nomoto, F., Sakoda, A. et al. Stereoselective synthesis of (R)-3-quinuclidinol through asymmetric reduction of 3-quinuclidinone with 3-quinuclidinone reductase of Rhodotorula rubra . Appl Microbiol Biotechnol 83, 617–626 (2009). https://doi.org/10.1007/s00253-009-1902-2
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DOI: https://doi.org/10.1007/s00253-009-1902-2