The hydrogen cycle in nitrogen-fixing Azotobacter chroococcum.

H2 will support nitrogenase activity (C2H2 reduction) in Azotobacter chroococcum with or without added carbon substrate. Results show that H2 is metabolised to transfer electrons to nitrogenase and to the respiratory chain to produce ATP. H2-supported nitrogenase activity is most significant at low carbon substrate concentrations, but also occurs at saturating concentration. Continuous cultures of N2-fixing A. chroococcum evolved H2 from nitrogenase under O2-N2- and C-limited conditions. This H2 represented a significant proportion of nitrogenase activity. Hydrogenase activity was consistently high under C-limited conditions, but low or undetectable under O2- and N2-limitations. Pre-treatment with 40 per cent C2H2 inhibited hydrogenase activity in C-limited cultures, and H2 evolution increased under air and under Ar:O2 (4:1) mixtures. We deduce that hydrogenase : I, recycles H2 produced by nitrogenase to provide electrons and energy for N2 reduction: II, supports respiratory protection for nitrogenase under C-limited conditions, and III, does not act to prevent any inhibition of N2 reduction by H2 produced by nitrogenase. A scheme for the H2 cycle in N2-fixing A. chroococcum is proposed.