Expression, purification, crystallization and preliminary X-ray diffraction analysis of carbonyl reductase from Candida parapsilosis ATCC 7330

被引：12

作者：

Aggarwal, Nidhi ^{[1
]}

Mandal, P. K. ^{[2
]}

Gautham, Namasivayam ^{[2
]}

Chadha, Anju ^{[1
,3
]}

机构：

[1] Indian Inst Technol, Dept Biotechnol, Madras 600036, Tamil Nadu, India

[2] Univ Madras, CAS Crystallog & Biophys, Madras 600025, Tamil Nadu, India

[3] Indian Inst Technol, Natl Ctr Catalysis Res, Madras 600036, Tamil Nadu, India

来源：

ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS | 2013年 / 69卷

关键词：

carbonyl reductase; alcohol dehydrogenase; enantioselectivity; Candida parapsilosis; ALCOHOL-DEHYDROGENASE; ASYMMETRIC REDUCTION;

D O I：

10.1107/S1744309113003667

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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 angstrom 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.

引用

页码：313 / 315

页数：3