Addition of 2 mM nitrite or ammonium to aerobically incubated cultures
of Gloeothece rapidly inhibited N2
fixation (measured as acetylene reduction). In contrast, 2 mM nitrate inhibited
N2 fixation less rapidly and less
extensively, and often temporarily stimulated nitrogenase activity. The
inhibitory effects of both nitrate and
ammonium could be prevented by addition of 3 mM L-methionine-DL-sulphoximine,
suggesting that the true
inhibitor of N2 fixation was an assimilatory product of ammonium
rather than either ammonium or nitrate itself.
The inhibition of N2 fixation by nitrite could not, however,
be prevented by addition of L-methionine-DL-
sulphoximine. On the other hand, nitrite (unlike nitrate and ammonium)
did not inhibit N2 fixation in cultures
incubated under a gas phase lacking oxygen. These findings suggest that
the mechanism whereby nitrite inhibits
N2 fixation in Gloeothece differs from that of either
nitrate or ammonium. The inhibitory effect of nitrite on N2
fixation did not involve reduction of nitrite to nitric oxide, though nitric
oxide was a potent inhibitor of nitrogenase
activity in Gloeothece. Nitrate and nitrite inhibited the synthesis
of nitrogenase in Gloeothece, while ammonium not
only inhibited nitrogenase synthesis but also stimulated degradation of
the enzyme. In addition, all three
compounds favoured the appearance of the Fe-protein of nitrogenase in its
larger, presumed inactive, form.