The long-term goal of this program is to engineer self-replicating bacterial systems to efficiently absorb sunlight and use the energy to split water and produce hydrogen. This crucial subproject is designed to provide fundamental insights required for the design of electron transfer pathways generally as well as the engineering and evolution of a specific photosynthetic hydrogen-producing pathway. The project will develop and apply methods that enable the evolution of a fusion protein that docks at photosystem I on a thylakoid membrane. The fusion protein will also be engineered and evolved to efficiently transfer electrons from the photosystem to an [FeFe] hydrogenase. The specific aims include (i)development of a high-throughput in vitro screen for hydrogen production driven by photosynthesis; (ii)engineering of a ferredoxin-hydrogenase fusion enzyme that produces hydrogen by coupling to photosystem I and (iii) the characterization of the productivity and oxygen sensitivity of the engineered photosynthetic hydrogen production system. The broader impacts are significant both for training and technological applications. It will teach collaboration, teamwork and communication skills to a diverse group of students including minorities. The broader application of the principles revealed by this program could reshape the energy and chemical industries and help to provide a sustainable global socio-economic system.
