europaea. In contrast to exponential phase, the statistical increase in relative mRNA concentrations with increasing DO concentrations for all four genes during stationary phase VX-661 molecular weight is clearly intriguing. These trends highlight the impact of starvation on responses to different DO concentrations. Although the unique responses of N. europaea to starvation [23] and oxygen concentrations (via Fnr [26]) have been documented, the mechanisms of combined NH3 and DO based gene regulation in N. europaea are not well understood. It is well documented that ammonia oxidizing bacteria, such as N. europaea, are commonly subject to cycling between anoxic and oxic conditions and

a wide range of NH3 concentrations in engineered and natural environments such as wastewater treatment plants or soils [24, 27, 28].

The specific responses observed herein might be part of a coordinated strategy of N. europaea to maintain active or latent substrate metabolic machinery to counter such varying environments and clearly merit further study. The differences in observed transient accumulation of NH2OH could also be explained at the transcription and protein activity levels. The decrease in exponential phase hao relative mRNA concentrations with increasing DO was more rapid than for amoA (Figure 3 A4-C4). This decrease coupled with a decrease in sOUR (a composite measure of AMO and HAO activity) with increasing DO, could have resulted in the observed trends in NH2OH concentrations. Although it has been shown that N. europaea can retain high levels of HAO protein and activity under ammonia starvation [29], the impact of DO concentrations on HAO activity has not been specifically identified. While check details the gene transcript data provide good insights into possible responses of N. europaea to different DO concentrations, protein activity data is crucial to explain profiles of intermediates

such as NH2OH. The AZD1152 concentration parallel profiles of exponential phase enough nirK relative mRNA concentrations and headspace NO concentrations at different DO concentrations (Figure 3) suggest a possible link between nirK transcription and NO generation. However, the loss of this parallel in the presence of added NO2 – (higher nirK gene transcription but lower NO concentrations, Figure 4) suggests the possible presence of NO generation pathways that are distinct from NO2 – reduction, as pointed out previously [26] or even post-transcriptional effects. Indeed, there is still no consensus about the source of NO produced by AOB, such as N. europaea, and the potential roles of nirK, hao and a multicopper oxidase of the nirK operon have all been implicated [26]. Impact of exposure to high nitrite concentrations on gene transcription High NO2 – concentrations have been implicated as the principal trigger for high NirK protein activity in N. europaea [9], which has a fundamental grounding in the similar trends observed in this study at the nirK gene mRNA level during exponential growth (Figure 4 D4).