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Towards the optimization of hydrogen production by nitrogenase (TOHPN)
Start date: 01 Oct 2008, End date: 30 Sep 2014 PROJECT  FINISHED 

In nature, molecular hydrogen is produced by the hydrogenase and the nitrogenase enzymes. Nitrogenase reduces dinitrogen to ammonia and, in this process, it evolves hydrogen. Nitrogenase and hydrogenase are oxygen-sensitive enzymes. We chose to optimize a hydrogen production system based on nitrogenase for four reasons: some organisms carrying nitrogenase simultaneously perform photosynthesis and hydrogen evolution by nitrogenase (direct biophotolysis), thus harvesting solar energy and autonomously converting it into chemical energy in a continuous process; cellular mechanisms exist to protect nitrogenase from oxygen but do not appear to exist for hydrogenase; because nitrogenase couples ATP hydrolysis to hydrogen evolution, this enzyme is able to generate hydrogen against a substantial gas pressure; finally, the biochemistry of the nitrogenase system is well known. The objective of our proposal is to provide new eco-efficient strategies for the biological production of hydrogen. Energy research is a priority theme under the Seventh Research Framework (FP7) cooperation program. The objective of energy research under FP7 is to adapt the current energy system into a more sustainable, competitive and secure one, with emphasis and support given to hydrogen research and renewable fuel production. Our proposal has three major components: (i) in vitro evolution of nitrogenase, in which we generate new nitrogenase variants by metagenomic gene shuffling and random mutagenesis, and select those with increased hydrogen production activity; (ii) the development of a genetic system to select for hydrogen overproducers; and (iii) a biochemical element designed to understand the biochemical requisites for efficient hydrogen production by the molybdenum nitrogenase as a basis for its re-engineering.
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