Nitric oxide is an obligate bacterial nitrification intermediate produced by hydroxylamine oxidoreductase

Ammonia (NH₃)-oxidizing bacteria (AOB) emit substantial amounts of nitric oxide (NO) and nitrous oxide (N₂O), both of which contribute to the harmful environmental side effects of large-scale agriculture. The currently accepted model for AOB metabolism involves NH₃ oxidation to nitrite (NO₂⁻) via a single obligate intermediate, hydroxylamine (NH₂OH). Within this model, the multiheme enzyme hydroxylamine oxidoreductase (HAO) catalyzes the four-electron oxidation of NH₂OH to NO₂⁻ We provide evidence that HAO oxidizes NH₂OH by only three electrons to NO under both anaerobic and aerobic conditions. NO₂⁻ observed in HAO activity assays is a nonenzymatic product resulting from the oxidation of NO by O₂ under aerobic conditions. Our present study implies that aerobic NH₃ oxidation by AOB occurs via two obligate intermediates, NH₂OH and NO, necessitating a mediator of the third enzymatic step.