title: Chemical assembly of multiple metal cofactors: The heterologously expressed multidomain [FeFe]-hydrogenase from Megasphaera elsdenii.
abstract: [FeFe]-hydrogenases are unique and fascinating enzymes catalyzing the reversible reduction of protons into hydrogen. These metalloenzymes display extremely large catalytic reaction rates at very low overpotential values and are, therefore, studied as potential catalysts for bioelectrodes of electrolyzers and fuel cells. Since they contain multiple metal cofactors whose biosynthesis depends on complex protein machineries, their preparation is difficult. As a consequence still few have been purified to homogeneity allowing spectroscopic and structural characterization. As part of a program aiming at getting easy access to new hydrogenases we report here a methodology based on a purely chemical assembly of their metal cofactors. This methodology is applied to the preparation and characterization of the hydrogenase from the fermentative anaerobic rumen bacterium Megasphaera elsdenii, which has only been incompletely characterized in the past.
authors.affiliation: Laboratoire de Chimie des Processus Biologiques, Collège de France, Université Pierre et Marie Curie, CNRS UMR 8229, 11 place Marcelin Berthelot, 75005 Paris, France.
authors.forename: Giorgio
authors.initials: G
authors.lastname: Caserta
authors.affiliation: Max-Planck-Institut für Chemische Energiekonversion, Stiftstrasse 34-36, 45470 Mülheim an der Ruhr, Germany.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Adamska-Venkatesh
authors.affiliation: Laboratoire de Chimie des Processus Biologiques, Collège de France, Université Pierre et Marie Curie, CNRS UMR 8229, 11 place Marcelin Berthelot, 75005 Paris, France.
authors.forename: Ludovic
authors.initials: L
authors.lastname: Pecqueur
authors.affiliation: Laboratoire de Chimie et Biologie des Métaux, Université Grenoble Alpes, CEA/BIG, CNRS, 17 rue des martyrs, 38000 Grenoble, France.
authors.forename: Mohamed
authors.initials: M
authors.lastname: Atta
authors.affiliation: Laboratoire de Chimie et Biologie des Métaux, Université Grenoble Alpes, CEA/BIG, CNRS, 17 rue des martyrs, 38000 Grenoble, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Artero
authors.affiliation: Laboratoire de Chimie et Biologie des Métaux, Université Grenoble Alpes, CEA/BIG, CNRS, 17 rue des martyrs, 38000 Grenoble, France.
authors.forename: Souvik
authors.initials: S
authors.lastname: Roy
authors.affiliation: Max-Planck-Institut für Chemische Energiekonversion, Stiftstrasse 34-36, 45470 Mülheim an der Ruhr, Germany.
authors.forename: Edward
authors.initials: E
authors.lastname: Reijerse
authors.affiliation: Max-Planck-Institut für Chemische Energiekonversion, Stiftstrasse 34-36, 45470 Mülheim an der Ruhr, Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Lubitz
authors.affiliation: Laboratoire de Chimie des Processus Biologiques, Collège de France, Université Pierre et Marie Curie, CNRS UMR 8229, 11 place Marcelin Berthelot, 75005 Paris, France. Electronic address: mfontecave@cea.fr.
authors.forename: Marc
authors.initials: M
authors.lastname: Fontecave
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D003067:Coenzymes / Q000737:chemistry* / Q000378:metabolism; D006864:Hydrogenase / Q000737:chemistry* / Q000378:metabolism; D007501:Iron / Q000737:chemistry / Q000378:metabolism*; D000070101:Megasphaera elsdenii / Q000201:enzymology*; D056004:Molecular Dynamics Simulation; D011485:Protein Binding; D055503:Protein Multimerization
chemical_list: D001426:Bacterial Proteins; D003067:Coenzymes; D007501:Iron; D006864:Hydrogenase
keywords: H(2) production; Megasphaera elsdenii; Synthetic maturation; [FeFe]-hydrogenase