Oxidation of ammonia by Nitrosomonas europaea. Definite 18O-tracer evidence that hydroxylamine formation involves a monooxygenase

Abstract

NH2OH, the first intermediate in the oxidation of NH4+ to nitrite by the nitrifying bacterium, Nitrosomonas europaea, was recovered as the oxime of cyclohexanone. 15N, 18O-tracer experiments using highly enriched 15NH4Cl and 18O2 yielded oxime that was correspondingly highly enriched (greater than or equal to 92 atom %) in these isotopes. These results show that the source of NH2OH is largely or entirely NH4+, as opposed to hydrazine, which was added to inhibit the further oxidation of NH2OH to nitrite, and that NH4+ yields NH2OH by way of a monooxygenase reaction involving direct insertion of O from O2. The oxidation of NH4+ and NH2OH must be functionally linked in N. europaea, inasmuch as the reducing equivalents required by the monooxygenase to reduce the second atom of O2 to water can arise only through the concomitant oxidation of NH2OH.