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砖红镰孢菌氰化物水合酶在大肠杆菌中的表达及一个必需半胱氨酸残基的鉴定

Expression of the cyanide hydratase enzyme from Fusarium lateritium in Escherichia coli and identification of an essential cysteine residue.

作者信息

Brown D T, Turner P D, O'Reilly C

机构信息

School of Health Sciences, University of Sunderland, UK.

出版信息

FEMS Microbiol Lett. 1995 Dec 15;134(2-3):143-6. doi: 10.1111/j.1574-6968.1995.tb07928.x.

Abstract

The filamentous fungus Fusarium lateritium is cyanide tolerant, due partly to the induction of the enzyme cyanide hydratase in the presence of cyanide. This enzyme catalyses the hydration of cyanide to formamide. The expression in Escherichia coli of a cDNA clone encoding cyanide hydratase is described. The cDNA cloned was expressed as a transcriptional fusion in the expression vector pKK233-2 and a high level of activity of cyanide hydratase was detected in E. coli. Site-directed mutagenesis of the cys-163 residue inactivated the enzyme.

摘要

丝状真菌砖红镰孢菌对氰化物具有耐受性,部分原因是在氰化物存在的情况下会诱导氰化物水合酶的产生。这种酶催化氰化物水合形成甲酰胺。本文描述了编码氰化物水合酶的cDNA克隆在大肠杆菌中的表达。克隆的cDNA在表达载体pKK233-2中作为转录融合体表达,并且在大肠杆菌中检测到高水平的氰化物水合酶活性。对cys-163残基进行定点诱变使该酶失活。

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