Homologous and heterologous overexpression in Clostridium acetobutylicum and characterization of purified clostridial and algal Fe-only hydrogenases with high specific activities

被引：102

作者：

Girbal, L

von Abendroth, G

Winkler, M

Benton, PMC

Meynial-Salles, I

Croux, C

Peters, JW

Happe, T

Soucaille, P

机构：

[1] CNRS, UMR 5504, Lab Biotechnol Bioproc, UMR 792,INSA, F-31077 Toulouse, France

[2] INSA, CRT, CRITT Bioind, DGBA, F-31077 Toulouse, France

[3] Montana State Univ, Dept Chem & Biochem, Bozeman, MT 59717 USA

[4] Ruhr Univ Bochum, AG Photobiotechnol, Lehrstuhl Biochem Pflanzen, D-44780 Bochum, Germany

来源：

APPLIED AND ENVIRONMENTAL MICROBIOLOGY | 2005年 / 71卷 / 05期

关键词：

D O I：

10.1128/AEM.71.5.2777-2781.2005

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Clostridium acetobutylicum ATCC 824 was selected for the homologous overexpression of its Fe-only hydrogenase and for the heterologous expressions of the Chlamydomonas reinhardtii and Scenedesmus obliquus HydA1 Fe-only hydrogenases. The three Strep tag H-tagged Fe-only hydrogenases were isolated with high specific activities by two-step column chromatography. The purified algal hydrogenases evolve hydrogen with rates of around 700 mu mol H-2 min(-1) mg(-1), while HydA from C. acetobutylicum (HydA(Ca)) shows the highest activity (5,522 mu mol H-2 min(-1) mg(-1)) in the direction of hydrogen uptake. Further, kinetic parameters and substrate specificity were reported. An electron paramagnetic resonance (EPR) analysis of the thionin-oxidized HydA(Ca) protein indicates a characteristic rhombic EPR signal that is typical for the oxidized H cluster of Fe-only hydrogenases.

引用

页码：2777 / 2781

页数：5

共 29 条

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