Expression, purification, crystallization and X‐ray analysis of 3‐quinuclidinone reductase from
Abstract:(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 (V M = 2.08 Å3 Da−1).
Document Type: Research Article
Publication date: October 1, 2012