Full metadata
Title
Engineering Metalloproteins for Solar Driven Hydrogen Production
Description
Hydrogenase enzymes capable of catalyzing proton reduction to produce H2 have generated a considerable interest due to increasing motivation in finding sustainable carbon free energy sources. A considerable amount of research has been focused on producing synthetic structures mimicking the hydrogenase catalytic site, but the activity seen in hydrogenase enzymes in aqueous near neutral pH has yet to be replicated. It is now clear that the protein structure surrounding the H-cluster enables the high activity by fine tuning characteristics of the catalyst, but the structure and complexity of hydrogenase enzymes makes it difficult to predict exactly how the secondary coordination sphere affects catalysis. This work looks at incorporating both synthetic molecular catalysts and hydrogenase mimics into peptide scaffolds to improve the activity for photo-driven H2 production in aqueous solutions. The first chapter of this dissertation shows a de novo heme binding peptide improving the activity of cobalt protoporphyrin IX (CoPPIX) upon coordination inside a four-helix bundle. The peptide bound CoPPIX exhibited a 5.5-fold increase in anaerobic and an 8.3-fold increase in aerobic activity compared to free CoPPIX, while also showing dramatic increases to stability and solubility. In the second chapter, this work is expanded by using a randomly mutated cytochrome b562 library to identify beneficial attributes for downstream implementation of an ideal coordination site. A high-throughput assay was developed to measure H2 production using WO3/Pd deposited on a glass plate for a colorimetric first-pass screen. This assay successfully measured H2 production from CoPPIX bound cytochrome b562 in the periplasm of cells and identified a possible mutant showing 70% more H2 production compared to the wildtype. The third chapter incorporated a hydrogenase mimic into a four-helix bundle using a semi-synthetic strategy yielding a 3-fold increase in activity due to catalyst encapsulation. The method created will allow for easy modifications to the synthetic catalyst or peptide sequence in future work. The systems developed in this work were designed to facilitate the identification and implementation of beneficial characteristics for future development of an optimal secondary coordination sphere for a peptide bound molecular catalyst.
Date Created
2021
Contributors
- Halloran, Nicholas Ryan (Author)
- Ghirlanda, Giovanna (Thesis advisor)
- Mills, Jeremy H (Committee member)
- Moore, Gary F (Committee member)
- Arizona State University (Publisher)
Topical Subject
Resource Type
Extent
125 pages
Language
eng
Copyright Statement
In Copyright
Primary Member of
Peer-reviewed
No
Open Access
No
Handle
https://hdl.handle.net/2286/R.2.N.161254
Level of coding
minimal
Cataloging Standards
Note
Partial requirement for: Ph.D., Arizona State University, 2021
Field of study: Biochemistry
System Created
- 2021-11-16 11:32:55
System Modified
- 2021-11-30 12:51:28
- 2 years 11 months ago
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