A simple hydrogenase-linked assay for ferredoxin and flavodoxin.

Ferredoxin or flavodoxin mediates electron flow from H₂-hydrogenase to metronidazole1-(2-hydroxyethyl)-2-methyl-5-nitroimidazole] to cause the reduction of the latter compound. The reduction of metronidazole in solution is irreversible because the reduced compound further decomposes. Since metronidazole loses its absorption peak at 320 nm upon reduction, the rate of reduction can be monitored spectrophotometrically. When a solution of metronidazole at 0.1 to 0.5 mm is supplemented with ferredoxin- andflavodoxin-free hydrogenase and placed under H₂, the rate of metronidazole reduction is proportional to the amount of ferredoxin or flavodoxin added. This forms the basis for an assay that can measure 10 to 1000 ng of ferredoxin or 100–1000 ng of flavodoxin/ml of assay mixture.