title: Expression, purification, crystallization and preliminary X-ray diffraction analysis of carbonyl reductase from Candida parapsilosis ATCC 7330.
abstract: The NAD(P)H-dependent carbonyl reductase from Candida parapsilosis ATCC 7330 catalyses the asymmetric reduction of ethyl 4-phenyl-2-oxobutanoate to ethyl (R)-4-phenyl-2-hydroxybutanoate, a precursor of angiotensin-converting enzyme inhibitors such as Cilazapril and Benazepril. The carbonyl reductase was expressed in Escherichia coli and purified by GST-affinity and size-exclusion chromatography. Crystals were obtained by the hanging-drop vapour-diffusion method and diffracted to 1.86 Å resolution. The asymmetric unit contained two molecules of carbonyl reductase, with a solvent content of 48%. The structure was solved by molecular replacement using cinnamyl alcohol dehydrogenase from Saccharomyces cerevisiae as a search model.
authors.affiliation: Department of Biotechnology, Indian Institute of Technology Madras, Chennai 600 036, India.
authors.forename: Nidhi
authors.initials: N
authors.lastname: Aggarwal
authors.forename: P K
authors.initials: PK
authors.lastname: Mandal
authors.forename: Namasivayam
authors.initials: N
authors.lastname: Gautham
authors.forename: Anju
authors.initials: A
authors.lastname: Chadha
mesh_terms: D000429:Alcohol Oxidoreductases / Q000737:chemistry* / Q000235:genetics; D000449:Aldehyde Reductase; D000074408:Aldo-Keto Reductases; D002175:Candida / Q000737:chemistry* / Q000201:enzymology; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000201:enzymology; D040681:Structural Homology, Protein
chemical_list: D005656:Fungal Proteins; D011994:Recombinant Proteins; D000429:Alcohol Oxidoreductases; D000074408:Aldo-Keto Reductases; C018656:cinnamyl alcohol dehydrogenase; D000449:Aldehyde Reductase
keywords: Candida parapsilosis; alcohol dehydrogenase; carbonyl reductase; enantioselectivity