Crystallization and preliminary X-ray analysis of the NADPH-dependent 3-quinuclidinone reductase from Rhodotorula rubra

• Published in: Acta crystallographica. Section F, Structural biology and crystallization communications Vol. 65; no. 6; pp. 645 - 647
• Main Authors: Takeshita, Daijiro, Kataoka, Michihiko, Miyakawa, Takuya, Miyazono, Ken-ichi, Uzura, Atsuko, Nagata, Koji, Shimizu, Sakayu, Tanokura, Masaru
• Format: Journal Article
• Language: English
• Published: 5 Abbey Square, Chester, Cheshire CH1 2HU, England International Union of Crystallography 01.06.2009
• Subjects: 3-quinuclidinone reductase; Alcohol Oxidoreductases - genetics; Alcohol Oxidoreductases - isolation & purification; Alcohol Oxidoreductases - metabolism; Aldehyde Reductase; Aldo-Keto Reductases; carbonyl reductases; Catalysis; Cloning, Molecular; Crystallization; Crystallization Communications; Crystallography, X-Ray; Data Collection; Escherichia coli; Escherichia coli - genetics; Genetic Vectors; Hydrogen-Ion Concentration; Plasmids; Quinuclidines - isolation & purification; Quinuclidines - metabolism; Rhodotorula - enzymology; Rhodotorula rubra; short-chain dehydrogenase/reductase family; Statistics as Topic; Substrate Specificity; Temperature; Transformation, Bacterial; X-Ray Diffraction
• ISSN: 1744-3091; 1744-3091
• DOI: 10.1107/S1744309109017588

(R)‐3‐Quinuclidinol is a useful compound that is applicable to the synthesis of various pharmaceuticals. The NADPH‐dependent carbonyl reductase 3‐quinuclidinone reductase from Rhodotorula rubra catalyzes the stereospecific reduction of 3‐quinuclidinone to (R)‐3‐quinuclidinol and is expected to be utilized in industrial production of this alcohol. 3‐Quinuclidinone reductase from R. rubra was expressed in Escherichia coli and purified using Ni‐affinity and ion‐exchange column chromatography. Crystals of the protein were obtained by the sitting‐drop vapour‐diffusion method using PEG 8000 as the precipitant. The crystals belonged to space group P41212, with unit‐cell parameters a = b = 91.3, c = 265.4 Å, and diffracted X‐rays to 2.2 Å resolution. The asymmetric unit contained four molecules of the protein and the solvent content was 48.4%.