Genetic and biochemical characterization of a short-chain alcohol dehydrogenase from the hyperthermophilic archaeon Pyrococcus furiosus

被引：41

作者：

van der Oost, J ^{[1
]}

Voorhorst, WGB ^{[1
]}

Kengen, SWM ^{[1
]}

Geerling, ACM ^{[1
]}

Wittenhorst, V ^{[1
]}

Gueguen, Y ^{[1
]}

de Vos, WM ^{[1
]}

机构：

[1] Univ Wageningen & Res Ctr, Microbiol Lab, NL-6703 CT Wageningen, Netherlands

来源：

EUROPEAN JOURNAL OF BIOCHEMISTRY | 2001年 / 268卷 / 10期

关键词：

alcohol dehydrogenase; Pyrococcus furiosus; short-chain; thermophilic;

D O I：

10.1046/j.1432-1327.2001.02201.x

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The gene encoding a short-chain alcohol dehydrogenase, AdhA, has been identified in the hyperthermophilic archaeon Pyrococcus furiosus, as part of an operon that encodes two glycosyl hydrolases, the beta -glucosidase CelB and the endoglucanase LamA. The adhA gene was functionally expressed in Escherichia coli, and AdhA was subsequently purified to homogeneity. The quaternary structure of AdhA is a dimer of identical 26-kDa subunits. AdhA is an NADPH-dependent oxidoreductase that converts alcohols to the corresponding aldehydes/ketones and vice versa, with a rather broad substrate specificity. Maximal specific activities were observed with 2-pentanol (46 U.mg(-1)) and pyruvaldehyde (32 U.mg(-1)) in the oxidative and reductive reaction, respectively. AdhA has an optimal activity at 90 degreesC, at which temperature it has a half life of 22.5 h. The expression of the adhA gene in P. furiosus was demonstrated by activity measurements and immunoblot analysis of cell extracts. A role of this novel type of archaeal alcohol dehydrogenase in carbohydrate fermentation is discussed.

引用

页码：3062 / 3068

页数：7

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