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Nitrate reductase from spinach (Spinacia oleracea L.) leaves, which had been inactivated in vitro by incubation with NADH and cyanide, was fully reactivated in minutes when irradiated in anaerobic conditions with red light in the presence of methylene blue. Both the rate and the extent of reactivation increased with light intensity (6 to 100 W·m -2 ) and dye concentration (1 to 10 μM). On the contrary, photoreactivation was completely abolished when NADH or ethylenediaminetetra-acetic acid were present during irradiation. We propose that methylene blue, when photo excited, exhibits a redox potential positive enough to reoxidise the CN - -re-duced molybdenum complex settled in the inactive enzyme, thus causing its reactivation.

Red-Light Effects Sensitized by Methylene Blue on Nitrate Reductase from Spinach (Spinacia oleracea L.) Leaves