Open AccessExpression, purification, crystallization and preliminary X‐ray analysis of carbonyl reductase S1 from Candida magnoliae
Author(s) -
Suwa Yoichi,
Ohtsuka Jun,
Miyakawa Takuya,
Imai Fabiana Lica,
Okai Masahiko,
Sawano Yoriko,
Yasohara Yoshihiko,
Kataoka Michihiko,
Shimizu Sakayu,
Tanokura Masaru
Publication year - 2012
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309112011645
Subject(s) - reductase , crystallization , solvent , chemistry , aldehyde reductase , crystallography , stereochemistry , molecule , enzyme , nuclear chemistry , organic chemistry
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 P 6 1 22 or P 6 5 22, 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%.