title: Expression, purification, crystallization and X-ray analysis of 3-quinuclidinone reductase from Agrobacterium tumefaciens.
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 P2(1), 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 Å(3) Da(-1)).
authors.affiliation: Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Feng
authors.initials: F
authors.lastname: Hou
authors.forename: Takuya
authors.initials: T
authors.lastname: Miyakawa
authors.forename: Daijiro
authors.initials: D
authors.lastname: Takeshita
authors.forename: Michihiko
authors.initials: M
authors.lastname: Kataoka
authors.forename: Atsuko
authors.initials: A
authors.lastname: Uzura
authors.forename: Koji
authors.initials: K
authors.lastname: Nagata
authors.forename: Sakayu
authors.initials: S
authors.lastname: Shimizu
authors.forename: Masaru
authors.initials: M
authors.lastname: Tanokura
mesh_terms: D016960:Agrobacterium tumefaciens / Q000201:enzymology*; D003460:Crystallization; D018360:Crystallography, X-Ray; D015870:Gene Expression; D011808:Quinone Reductases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011812:Quinuclidines / Q000378:metabolism
chemical_list: D011812:Quinuclidines; D011808:Quinone Reductases