Overview of the results:
1) The project has provided extensive training to the Fellow on innovative infrared techniques (FTIR) applied to metalloenzymes and a productive two-way transfer of knowledge with the host group.
2) Protein Film IR Electrochemistry (PFIRE) has been applied for the first time to [FeFe]-hydrogenases. The technique allows for simultaneous investigation of functional and spectroscopic features during catalytic turnover of redox enzymes. The results obtained show feasibility of this approach and set the basis for future more detailed investigation.
3) Custom-built spectro-electrochemical equipment allowed mechanistic investigation on the role of proton transfer within [FeFe]-hydrogenases, and how this influences the redox equilibria at the active site.
4) FTIR micro-spectroscopy was used to investigate how these enzymes respond when in the crystalline state. This work showed that the enzymes retain their natural reactivity when crystallised and that these features can easily be controlled during the experiment. This provides a powerful tool for controlling and determining the exact redox state of the sample under given conditions and opens completely new perspectives in structural biology.
Exploitation and dissemination:
The results have been presented to the following national meetings and international conferences:
- Hydrogenase workshop, Berlin, Germany, 2018.
- European Biological Inorganic Chemistry Conference (EuroBIC), Birmingham, UK, 2018.
- International Hydrogenase Conference in Lisbon, Portugal, 2019.
- Royal Society of Chemistry Dalton Division Southern Regional Meeting in Oxford, UK, 2019.
The Fellow is now preparing two manuscripts for publication in peer-reviewed scientific journals describing the results of the project.
