Expression, purification, crystallization and preliminary X‐ray analysis of carbonyl reductase S1 from
Authors: Suwa, Yoichi; Ohtsuka, Jun; Miyakawa, Takuya; Imai, Fabiana Lica; Okai, Masahiko; Sawano, Yoriko; Yasohara, Yoshihiko; Kataoka, Michihiko; Shimizu, Sakayu; Tanokura, Masaru
Source: Acta Crystallographica Section F, Volume 68, Number 5, 1 May 2012 , pp. 540-542(3)
Abstract:The NADPH‐dependent carbonyl reductase S1 from Candida magnoliae stereoselectively catalyzes the reduction of ethyl 4‐chloro‐3‐oxobutanoate (COBE) to ethyl (S)‐4‐chloro‐3‐hydroxybutanoate (CHBE), which is a chiral compound valuable as a building block for pharmaceuticals. Carbonyl reductase S1 was expressed in Escherichia coli and purified by Ni‐affinity, ion‐exchange and size‐exclusion chromatography. Crystals of carbonyl reductase S1 were obtained by the sitting‐drop vapour‐diffusion method using PEG 400 as a precipitant. X‐ray diffraction data were collected to 1.90 Å resolution using a synchrotron‐radiation source. The crystals belonged to space group P6122 or P6522, with unit‐cell parameters a = b = 77.7, c = 307.5 Å. The asymmetric unit contained two molecules of the protein, with a solvent content of 44.2%.
Document Type: Research Article
Publication date: May 1, 2012