Crystallization and preliminary X-ray diffraction analysis of (R)-carbonyl reductase from Candida parapsilosis.

ABSTRACT: 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 P2?2?2?, 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.

SUBMITTER: Wang S 

PROVIDER: S-EPMC4051541 | biostudies-literature | 2014 Jun

REPOSITORIES: biostudies-literature