Comparison of Nitrogen Oxide Metabolism among Diverse Ammonia-Oxidizing Bacteria

Ammonia-oxidizing bacteria (AOB) have well characterized genes that encode and express nitrite reductases (NIR) and nitric oxide reductases (NOR). However, the connection between presence or absence of these and other genes for nitrogen transformations with the physiological production of nitric oxide (NO) and nitrous oxide (N(2)O) has not been tested across AOB isolated from various trophic states, with diverse phylogeny, and with closed genomes. It is therefore unclear if genomic content for nitrogen oxide metabolism is predictive of net N(2)O production. Instantaneous microrespirometry experiments were utilized to measure NO and N(2)O emitted by AOB during active oxidation of ammonia (NH(3)) or hydroxylamine (NH(2)OH) and through a period of anoxia. This data was used in concert with genomic content and phylogeny to assess whether taxonomic factors were predictive of nitrogen oxide metabolism. Results showed that two oligotrophic AOB strains lacking annotated NOR-encoding genes released large quantities of NO and produced N(2)O abiologically at the onset of anoxia following NH(3)-oxidation. Furthermore, high concentrations of N(2)O were measured during active O(2)-dependent NH(2)OH oxidation by the two oligotrophic AOB in contrast to non-oligotrophic strains that only produced N(2)O at the onset of anoxia. Therefore, complete nitrifier denitrification did not occur in the two oligotrophic strains, but did occur in meso- and eutrophic strains, even in Nitrosomonas communis Nm2 that lacks an annotated NIR-encoding gene. Regardless of mechanism, all AOB strains produced measureable N(2)O under tested conditions. This work further confirms that AOB require NOR activity to enzymatically reduce NO to N(2)O in the nitrifier denitrification pathway, and also that abiotic reactions play an important role in N(2)O formation, in oligotrophic AOB lacking NOR activity.