STUDIES ON ACTIVITY OF UPTAKE HYDROGENASE IN FRANKIA ARIS : Optimization of Assay Procedures and Culture Conditions
D'Aloisio, Aimee A.
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The actinomycete Frankia contains an uptake hydrogenase which recycles part of the energy consumed from the evolution of hydrogen during nitrogen fixation by the nitrogenase complex. The uptake hydrogenase catalyzes oxidation of hydrogen to form protons, and the electrons from this reaction are shuttled along the electron transport chain resulting in the eventual production of ATP by the ATPase. A technique for measuring the activity of this enzyme involves using an amperometric assay. Modifications to this techniques were made to improve the precision and reliability of this method. The expression of active hydrogenase is influenced by the growth conditions of the Frankia. Experiments were performed to determine the effect of nickel addition and removal from the growth media and of the presence of carbon source on the growth and expression Qf active hydro~enase. Results of these tests were inconclusive.