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[NiFe] hydrogenases from the hyperthermophilic bacterium Aquifex aeolicus: properties, function, and phylogenetics

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Abstract

Genes potentially coding for three distinct [NiFe] hydrogenases are present in the genome of Aquifex aeolicus. We have demonstrated that all three hydrogenases are expressed under standard growth conditions of the organism. Two hydrogenases were further purified to homogeneity. A periplasmically oriented hydrogenase was obtained in two forms, i.e., as a soluble enzyme containing only the two essential subunits and as a detergent-solubilized complex additionally containing a membrane-integral b-type cytochrome. The second hydrogenase purified was identified as a soluble cytoplasmic enzyme. The isolated enzymes were characterized with respect to biochemical/biophysical parameters, activity, thermostability, and substrate specificity. The phylogenetic positioning of all three hydrogenases was analyzed. A model for the metabolic roles of the three enzymes is proposed on the basis of the obtained results.

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Acknowledgements

We would like to thank Prof. T. Friedrich (Freiburg, Germany) for stimulating discussions, Dr. F. Baymann and Dr. C. Aubert (Marseille, France) for critical reading of the manuscript, as well as Dr. M. Rousset (Marseille, France) for valuable comments, D. Moinier (Protein sequencing Unit, B.I.P., Marseilles, France) for proteomic experiments, and R. Lebrun (Protein sequencing Unit, B.I.P., Marseilles, France) for performing the N-terminal sequence determination. The work of KOS was financially supported by the Fonds der Chemischen Industrie, and the project on Aquifex at the BIP in Marseille benefited from financial support by the PCV program of the CNRS and by the CNES.

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Authors and Affiliations

  1. Bioénergétique et Ingénierie des Protéines, CNRS, IBSM, 31 chemin Joseph Aiguier, 13402 , Marseille Cedex 20, France

    Marianne Brugna-Guiral, Pascale Tron, Wolfgang Nitschke, Benedicte Burlat, Bruno Guigliarelli, Mireille Bruschi & Marie Thérèse Giudici-Orticoni

  2. Lehrstuhl für Mikrobiologie, Universität Regensburg, Regensburg, Germany

    Karl-Otto Stetter

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  1. Marianne Brugna-Guiral
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  2. Pascale Tron
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  3. Wolfgang Nitschke
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  5. Benedicte Burlat
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Correspondence to Marie Thérèse Giudici-Orticoni.

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Brugna-Guiral, M., Tron, P., Nitschke, W. et al. [NiFe] hydrogenases from the hyperthermophilic bacterium Aquifex aeolicus: properties, function, and phylogenetics. Extremophiles 7, 145–157 (2003). https://doi.org/10.1007/s00792-002-0306-3

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