Expression, purification, crystallization and X-ray analysis of 3-quinuclidinone reductase from Agrobacterium tumefaciens

• Published in: Acta crystallographica. Section F, Structural biology and crystallization communications Vol. 68; no. 10; pp. 1237 - 1239
• Main Authors: Hou, Feng, Miyakawa, Takuya, Takeshita, Daijiro, Kataoka, Michihiko, 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 01.10.2012
• Subjects: 3-quinuclidinone reductase; Agrobacterium tumefaciens; Agrobacterium tumefaciens - enzymology; Asymmetry; Crystallization; Crystallization Communications; Crystallography, X-Ray; Crystals; Escherichia coli; Gene Expression; NADH; Quinone Reductases - chemistry; Quinone Reductases - genetics; Quinone Reductases - isolation & purification; Quinone Reductases - metabolism; Quinuclidines - metabolism; Reductases; Reduction; X-rays
• ISSN: 1744-3091; 1744-3091
• DOI: 10.1107/S1744309112034951

(R)‐3‐Quinuclidinol is a useful chiral building block for the synthesis of various pharmaceuticals and can be produced from 3‐quinuclidinone by asymmetric reduction. A novel 3‐quinuclidinone reductase from Agrobacterium tumefaciens (AtQR) catalyzes the stereospecific reduction of 3‐quinuclidinone to (R)‐3‐quinuclidinol with NADH as a cofactor. Recombinant AtQR was overexpressed in Escherichia coli, purified and crystallized with NADH using the sitting‐drop vapour‐diffusion method at 293 K. Crystals were obtained using a reservoir solution containing PEG 3350 as a precipitant. X‐ray diffraction data were collected to 1.72 Å resolution on beamline BL‐5A at the Photon Factory. The crystal belonged to space group P21, with unit‐cell parameters a = 62.0, b = 126.4, c = 62.0 Å, β = 110.5°, and was suggested to contain four molecules in the asymmetric unit (VM = 2.08 Å3 Da−1).