Analysis and Engineering of a novel CO linked O2 tolerant hydrogenase in photosynthetic bacteria

Hydrogen is a clean and renewable energy source that is naturally produced by a number of microorganisms via hydrogenase enzymes. Many hydrogenases are found in photosynthetic organisms and are utilized as energy sinks, but to harness their power as hydrogen producers, we must understand more about their role in the cell and their innate sensitivity to oxygen. Almost 20 years ago, an interesting purple non-sulfur photosynthetic bacterium, Rubrivivax gelatinous CBS (Casa Bonita Strain), was discovered in soil samples and was found to contain a CO-linked hydrogenase that exhibited a half-life of 21 hours in oxygen. Here, we will present data uncovering the intricate CO and hydrogen metabolism of this organism, our engineering of improved hydrogen production, and our attempts to express this hydrogenase and its maturation factors for function in other microbes.