Immobilization of the [FeFe]-hydrogenase CrHydA1 on a gold electrode: Design of a catalytic surface for the production of molecular hydrogen

• Published in: Journal of biotechnology 2009-06, Vol.142 (1), p.3-9
• Main Authors: Krassen, Henning, Stripp, Sven, von Abendroth, Gregory, Ataka, Kenichi, Happe, Thomas, Heberle, Joachim
• Format: Article
• Language: English
• Subjects: [FeFe]-hydrogenase; Adsorption; Animals; biofuels; Biological hydrogen production; Bioreactors; Chlamydomonas reinhardtii; Chlamydomonas reinhardtii - enzymology; Clostridium acetobutylicum - genetics; Electrodes; Electron transfer; enzyme activity; Enzyme-modified electrode; Enzymes, Immobilized - chemistry; Enzymes, Immobilized - genetics; Enzymes, Immobilized - metabolism; ferredoxin hydrogenase; Freshwater; FTIR; gold; Gold - chemistry; Hydrogen - metabolism; hydrogen production; Hydrogenase - chemistry; Hydrogenase - genetics; Hydrogenase - metabolism; immobilized enzymes; Iron-Sulfur Proteins - chemistry; Iron-Sulfur Proteins - genetics; Iron-Sulfur Proteins - metabolism; Kinetics; Monolayer; Recombinant Proteins - chemistry; Recombinant Proteins - metabolism; SEIRA; Spectroscopy, Fourier Transform Infrared; Surface Plasmon Resonance; Surface Properties
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/j.jbiotec.2009.01.018

Hydrogenase-modified electrodes are a promising catalytic surface for the electrolysis of water with an overpotential close to zero. The [FeFe]-hydrogenase CrHydA1 from the photosynthetic green alga Chlamydomonas reinhardtii is the smallest [FeFe]-hydrogenase known and exhibits an extraordinary high hydrogen evolution activity. For the first time, we immobilized CrHydA1 on a gold surface which was modified by different carboxy-terminated self-assembled monolayers. The immobilization was in situ monitored by surface-enhanced infrared spectroscopy. In the presence of the electron mediator methyl viologen the electron transfer from the electrode to the hydrogenase was detected by cyclic voltammetry. The hydrogen evolution potential (−290 mV vs NHE, pH 6.8) of this protein modified electrode is close to the value for bare platinum (−270 mV vs NHE). The surface coverage by CrHydA1 was determined to 2.25 ng mm −2 by surface plasmon resonance, which is consistent with the formation of a protein monolayer. Hydrogen evolution was quantified by gas chromatography and the specific hydrogen evolution activity of surface-bound CrHydA1 was calculated to 1.3 μmol H 2 min −1 mg −1 (or 85 mol H 2 min −1 mol −1). In conclusion, a viable hydrogen-evolving surface was developed that may be employed in combination with immobilized photosystems to provide a platform for hydrogen production from water and solar energy with enzymes as catalysts.