Tailor-Made Modification of a Gold Surface for the Chemical Binding of a High-Activity [FeFe] Hydrogenase

Authors

  • Henning Krassen,

    1. Bielefeld University, Department of Chemistry, Biophysical Chemistry, 33615 Bielefeld, Germany
    2. Uppsala University, Department of Photochemistry andMolecular Science, 75120 Uppsala, Sweden
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  • Sven T. Stripp,

    1. Ruhr-Universität Bochum, Lehrstuhl Biochemie der Pflanzen, AG Photobiotechnologie, 44801 Bochum, Germany
    2. Freie Universität Berlin, Department of Physics, Experimental Molecular Biophysics, 14195 Berlin, Germany
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  • Nadine Böhm,

    1. Universität zu Köln, Department für Chemie, Institut fürOrganische Chemie, 50939 Cologne, Germany
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  • Albrecht Berkessel,

    1. Universität zu Köln, Department für Chemie, Institut fürOrganische Chemie, 50939 Cologne, Germany
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  • Thomas Happe,

    1. Ruhr-Universität Bochum, Lehrstuhl Biochemie der Pflanzen, AG Photobiotechnologie, 44801 Bochum, Germany
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  • Kenichi Ataka,

    1. Bielefeld University, Department of Chemistry, Biophysical Chemistry, 33615 Bielefeld, Germany
    2. Freie Universität Berlin, Department of Physics, Experimental Molecular Biophysics, 14195 Berlin, Germany
    3. Japan Science and Technology Agency, 102-0075, Tokyo, Japan
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  • Joachim Heberle

    1. Bielefeld University, Department of Chemistry, Biophysical Chemistry, 33615 Bielefeld, Germany
    2. Freie Universität Berlin, Department of Physics, Experimental Molecular Biophysics, 14195 Berlin, Germany
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Abstract

Hydrogenases are iron–sulfur proteins that catalyze hydrogen turnover in a wide range of microorganisms. Three different classes have been described, and among these [FeFe] hydrogenases are the most active in H2 evolution. Hydrogenases are redox enzymes that have been shown to exchange electrons with graphite and modified noble metal electrodes. Making use of the latter, diffusible electron carriers are required to enable redox catalysis, as proteins do not specifically bind to the electrode surface. Diffusion-limited electron transfer can be replaced by electron injection into immobilized hydrogenase by binding the redox mediator to the electrode surface. Here, we present the synthesis and spectroelectrochemical characterization of 1-(10-mercaptodecyl)-1′-benzyl-4,4′-bipyridinium dibromide (MBBP), which is used as redox-active linker. CrHydA1, the high-activity [FeFe] hydrogenase from Chlamydomonas reinhardtii, is immobilized on the linker-modified gold electrode. Each surface modification step is controlled in situ by surface-enhanced infrared absorption spectroscopy (SEIRAS). Functionality of the electrode–protein hybrid is demonstrated by recording the linker-supported current. The specificcatalytic rate of hydrogen evolution by CrHydA1 (2.9 μmol H2 min–1 mg–1 hydrogenase) promises a valuable approach for further optimization of this novel bioelectrical interface.

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