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Crystallization and preliminary X‐ray diffraction analysis of ( R )‐carbonyl reductase from Candida parapsilosis
Author(s) -
Wang Shanshan,
Nie Yao,
Yan Xu,
Ko TzuPing,
Huang ChunHsiang,
Chan HsiuChien,
Guo ReyTing,
Xiao Rong
Publication year - 2014
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x1400908x
Subject(s) - candida parapsilosis , orthorhombic crystal system , nad+ kinase , reductase , chemistry , stereochemistry , homotetramer , crystallization , crystallography , candida albicans , enzyme , crystal structure , biochemistry , biology , organic chemistry , microbiology and biotechnology , protein subunit , gene
The NADH‐dependent ( R )‐carbonyl reductase from Candida parapsilosis (RCR) catalyzes the asymmetric reduction of 2‐hydroxyacetophenone (HAP) to produce ( R )‐1‐phenyl‐1,2‐ethanediol [( R )‐PED], which is used as a versatile building block for the synthesis of pharmaceuticals and fine chemicals. To gain insight into the catalytic mechanism, the structures of complexes of RCR with ligands, including the coenzyme, are important. Here, the recombinant RCR protein was expressed and purified in Escherichia coli and was crystallized in the presence of NAD + . The crystals, which belonged to the orthorhombic space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 85.64, b = 106.11, c = 145.55 Å, were obtained by the sitting‐drop vapour‐diffusion method and diffracted to 2.15 Å resolution. Initial model building indicates that RCR forms a homotetramer, consistent with previous reports of medium‐chain‐type alcohol dehydrogenases.