Details

Autor(en) / Beteiligte

• Girbal, Laurence
• von Abendroth, Gregory
• Winkler, Martin
• Benton, Paul M. C
• Meynial-Salles, Isabelle
• Croux, Christian
• Peters, John W
• Happe, Thomas
• Soucaille, Philippe

Titel

Homologous and Heterologous Overexpression in Clostridium acetobutylicum and Characterization of Purified Clostridial and Algal Fe-Only Hydrogenases with High Specific Activities

Ist Teil von

• Applied and Environmental Microbiology, 2005-05, Vol.71 (5), p.2777-2781

Ort / Verlag

Washington, DC: American Society for Microbiology

Erscheinungsjahr

2005

Quelle

MEDLINE

Beschreibungen/Notizen

• 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 II-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 [micro]mol H₂ min⁻¹ mg⁻¹, while HydA from C. acetobutylicum (HydA[subscript Ca]) shows the highest activity (5,522 [micro]mol H₂ 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[subscript Ca] protein indicates a characteristic rhombic EPR signal that is typical for the oxidized H cluster of Fe-only hydrogenases.