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Journal of Bacteriology, February 2008, p. 843-850, Vol. 190, No. 3
0021-9193/08/$08.00+0     doi:10.1128/JB.01417-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Coupled Ferredoxin and Crotonyl Coenzyme A (CoA) Reduction with NADH Catalyzed by the Butyryl-CoA Dehydrogenase/Etf Complex from Clostridium kluyveri{triangledown} ,{dagger}

Fuli Li,1,{ddagger} Julia Hinderberger,1,{ddagger} Henning Seedorf,1 Jin Zhang,2 Wolfgang Buckel,2 and Rudolf K. Thauer1*

Max Planck Institute for Terrestrial Microbiology, D-35043 Marburg, Germany,1 Laboratorium für Mikrobiologie, Fachbereich Biologie, Philipps-Universität, D-35032 Marburg, Germany2

Received 31 August 2007/ Accepted 26 October 2007

Cell extracts of butyrate-forming clostridia have been shown to catalyze acetyl-coenzyme A (acetyl-CoA)- and ferredoxin-dependent formation of H2 from NADH. It has been proposed that these bacteria contain an NADH:ferredoxin oxidoreductase which is allosterically regulated by acetyl-CoA. We report here that ferredoxin reduction with NADH in cell extracts from Clostridium kluyveri is catalyzed by the butyryl-CoA dehydrogenase/Etf complex and that the acetyl-CoA dependence previously observed is due to the fact that the cell extracts catalyze the reduction of acetyl-CoA with NADH via crotonyl-CoA to butyryl-CoA. The cytoplasmic butyryl-CoA dehydrogenase complex was purified and is shown to couple the endergonic reduction of ferredoxin (E0' = –410 mV) with NADH (E0' = –320 mV) to the exergonic reduction of crotonyl-CoA to butyryl-CoA (E0' = –10 mV) with NADH. The stoichiometry of the fully coupled reaction is extrapolated to be as follows: 2 NADH + 1 oxidized ferredoxin + 1 crotonyl-CoA = 2 NAD+ + 1 ferredoxin reduced by two electrons + 1 butyryl-CoA. The implications of this finding for the energy metabolism of butyrate-forming anaerobes are discussed in the accompanying paper.

* Corresponding author. Mailing address: Max Planck Institute for Terrestrial Microbiology, Karl-von-Frisch-Straβe, D-35043 Marburg, Germany. Phone: 49 6421 178 101. Fax: 49 6421 178 109. E-mail: thauer{at}mpi-marburg.mpg.de

{triangledown} Published ahead of print on 9 November 2007.

{dagger} Dedicated to Karl Decker, Emeritus Professor of Biochemistry, University Freiburg, Freiburg, Germany.

{ddagger} F.L. and J.H. contributed equally to this work.

Journal of Bacteriology, February 2008, p. 843-850, Vol. 190, No. 3
0021-9193/08/$08.00+0     doi:10.1128/JB.01417-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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