Two catalytically distinct molybdenum-free dissimilatory nitrate reductases, a soluble periplasmic and a membranebound one, were isolated from the vanadate-reducing facultatively anaerobic bacterium Pseudomonas isachenkovii and purified to electrophoretic homogeneity. The enzymes did not contain molybdenum, the periplasmic enzyme contained vanadium, whereas the membrane-bound enzyme was vanadium-free. Both nitrate reductases lacked molybdenum cofactor. This fact was proved by reconstitution of the apoprotein of the nitrate reductase of Neurospora crassa nit-1 mutant. This is the first demonstration of molybdenum-free and molybdenum cofactorfree nitrate reductases.z 1998 Federation of European Biochemical Societies.
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