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首页> 外文期刊>Acta crystallographica, Section F. Structural biology and crystallization communications >Expression, purification, crystallization and preliminary X-ray diffraction analysis of carbonyl reductase from Candida parapsilosis ATCC 7330
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Expression, purification, crystallization and preliminary X-ray diffraction analysis of carbonyl reductase from Candida parapsilosis ATCC 7330

机译：副念珠菌ATCC 7330中羰基还原酶的表达，纯化，结晶和初步X射线衍射分析

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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.
机译：来自副念珠菌ATCC 7330的NAD（P）H依赖性羰基还原酶催化4-苯基-2-氧代丁酸乙酯不对称还原为（R）-4-苯基-2-羟基丁酸乙酯（血管紧张素转换酶抑制剂的前体）如西拉普利和贝那普利。羰基还原酶在大肠杆菌中表达，并通过GST亲和力和尺寸排阻色谱法纯化。通过悬滴蒸气扩散法获得晶体并衍射至1.86埃的分辨率。不对称单元包含两个分子的羰基还原酶，溶剂含量为48％。通过使用来自酿酒酵母的肉桂醇脱氢酶作为搜索模型，通过分子置换来解决结构。

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