Crystallization and preliminary X-ray analysis of the NADPH-dependent 3-quinuclidinone reductase from Rhodotorula rubra.
(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 P4(1)2(1)2, with unit-cell parameters a = b = 91.3, c = 265.4 A, and diffracted X-rays to 2.2 A resolution. The asymmetric unit contained four molecules of the protein and the solvent content was 48.4%.
Takeshita
Daijiro
D
Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
Kataoka
Michihiko
M
Miyakawa
Takuya
T
Miyazono
Ken-ichi
K
Uzura
Atsuko
A
Nagata
Koji
K
Shimizu
Sakayu
S
Tanokura
Masaru
M
D000429:Alcohol Oxidoreductases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000449:Aldehyde Reductase; D000074408:Aldo-Keto Reductases; D002384:Catalysis; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D003625:Data Collection; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D006863:Hydrogen-Ion Concentration; D010957:Plasmids; D011812:Quinuclidines / Q000302:isolation & purification / Q000378:metabolism*; D012248:Rhodotorula / Q000201:enzymology*; D013223:Statistics as Topic; D013379:Substrate Specificity; D013696:Temperature; D014169:Transformation, Bacterial; D014961:X-Ray Diffraction
D011812:Quinuclidines; C090998:3-quinuclidinone; D000429:Alcohol Oxidoreductases; D000074408:Aldo-Keto Reductases; D000449:Aldehyde Reductase