TY  - THES
AB  - Hydrogenases catalyze the reduction of protons to molecular hydrogen with outstanding efficiency. An electrode surface which is covered with active hydrogenase molecules becomes a promising alternative to platinum for electrochemical hydrogen production.
To immobilize the hydrogenase on the electrode, the gold surface was modified by heterobifunctional molecules. A thiol headgroup on one side allowed the binding to the gold surface and the formation of a self-assembled monolayer. The other side of the molecules provided a surface with a high affinity for the hydrogenase CrHydA1 from Chlamydomonas reinhardtii. With methylviologen as a soluble energy carrier, electrons were transferred from carboxy-terminated electrodes to CrHydA1 and conducted to the active site (H-cluster), where they reduce protons to molecular hydrogen. A combined approach of surface-enhanced infrared absorption spectroscopy, gas chromatography, and surface plasmon resonance allowed quantifying the hydrogen production on a molecular level. Hydrogen was produced with a rate of 85 mol H2 min-1 mol-1. On a 1'-benzyl-4,4'-bipyridinum (BBP)-terminated surface, the electrons were mediated by the monolayer and no soluble electron carrier was necessary to achieve a comparable hydrogen production rate (approximately 50 percent of the former system). The hydrogen evolution potential was determined to be -335 mV for the BBP-bound hydrogenase and -290 mV for the hydrogenase which was immobilized on a carboxy-terminated mercaptopropionic acid SAM. Therefore, both systems significantly reduce the hydrogen production overpotential and allow electrochemical hydrogen production at an energy level which is close to the commercially applied platinum electrodes (hydrogen evolution potential of -270 mV).
In order to couple hydrogen production and photosynthesis, photosystem I (PS1) from Synechocystis PCC 6803 and membrane-bound hydrogenase (MBH) from Ralstonia eutropha were bound to each other. To accomplish tight binding of both proteins the PS1 subunit PsaE was genetically fused to the C-terminal end of the small subunit of MBH, i.e. close to the electron acceptor site of MBH. This fusion protein spontaneously assembled with the PsaE-deletion mutant of PS1. Crucial for a high hydrogen evolution rate of the system is an efficient electron transfer between both proteins. To allow this measurement, the PsaE-deletion mutant of PS1 was immobilized on a Ni-NTA terminated monolayer via a genetically introduced His-tag. The specificity of the assembly of fusion protein and deletion mutant was verified by SEIRAS. Surface plasmon resonance, gas chromatography and electrochemistry complemented this measurement and yielded the specific activity of the functional hybrid complex: 4500 mol H2 min-1 mol-1. The investigated complex allowed hydrogen evolution at potentials up to 85 mV, i.e. hydrogen production at a lower energy level than on a platinum electrode. In addition, the hydrogen production rate was higher than for hydrogenase-modified electrodes without PS1.
Beyond these specific results, the experimental setup can be used to quantify the hydrogen evolution rate on a molecular level for variable hydrogenases and hybrid complexes. This information will be used to choose the most efficient catalysts for introduction into the native system for in vivo hydrogen production.
DA  - 2009
KW  - Wasserstofferzeugung , Biokonversion , Wasserstoff , Hydrogenase , Infrarotspektroskopie , SEIRAS , Hydrogen , Infrared spectroscopy
LA  - eng
PY  - 2009
TI  - Biomimetic hydrogen production
UR  - https://nbn-resolving.org/urn:nbn:de:hbz:361-15245
Y2  - 2024-12-25T07:24:14
ER  -