Kinetics of leaf nitrite reductase with Methyl Viologen and ferredoxin under controlled redox conditions.

Some factors that influence the activity of nitrite reductase (EC 1.7.7.1) were investigated, the enzyme from Curcurbita pepo (vegetable marrow) being used. The activity with ferredoxin or Methyl Viologen as electron donor was inhibited by certain salts, including NaCl. The steady-state kinetic parameters measured in a commonly used open-tube (aerobic) system were compared with a closed-cell (anaerobic) system in which the redox potential, and thus the concentrations of oxidized and reduced donor, could be controlled. This showed that in the open-tube system the apparent Km values determined were overestimated (by a factor of 10 for reduced Methyl Viologen), owing to incomplete mediator reduction and competitive inhibition by the oxidized form of the mediator.