Stereoselective synthesis of (R)-3-quinuclidinol through asymmetric reduction of 3-quinuclidinone with 3-quinuclidinone reductase of Rhodotorula rubra

• Published in: Applied microbiology and biotechnology Vol. 83; no. 4; pp. 617 - 626
• Main Authors: Uzura, Atsuko, Nomoto, Fumiki, Sakoda, Akiko, Nishimoto, Yukifumi, Kataoka, Michihiko, Shimizu, Sakayu
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Berlin/Heidelberg : Springer-Verlag 01.06.2009; Springer Berlin Heidelberg; Springer; Springer Nature B.V
• Subjects: Alcohol Oxidoreductases - genetics; Alcohol Oxidoreductases - isolation & purification; Alcohol Oxidoreductases - metabolism; Amino Acid Sequence; Amino acids; Base Sequence; Biological and medical sciences; Biotechnological Products and Process Engineering; Biotechnology; Biotransformation; Carbonyl compounds; Chemical synthesis; Chromatography; Cloning, Molecular; Dehydrogenase; Dehydrogenases; DNA, Fungal - chemistry; DNA, Fungal - genetics; E coli; Enzymes; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Fungal Proteins - genetics; Fungal Proteins - isolation & purification; Fungal Proteins - metabolism; Gene Expression; Genes; Glucose; Glucose 1-Dehydrogenase - genetics; Glucose 1-Dehydrogenase - metabolism; Life Sciences; Membrane filters; Microbial Genetics and Genomics; Microbiology; Molecular Sequence Data; Molecular weight; Quinuclidines - metabolism; Rhodotorula - enzymology; Rhodotorula - genetics; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Stereoisomerism; Studies; Yeast; Asymmetric reduction; (; 3-Quinuclidinone reductase; 3-Quinuclidinol; Yeast; Enzyme; (R)-3-Quinuclidinol; Rhodotorularubra; Fungi; Stereoselectivity; Reduction; Fungi Imperfecti; Oxidoreductases; Rhodotorula rubra; Reductase
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-009-1902-2

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.