Nitrite Control over Dissimilatory Nitrate/Nitrite Reduction Pathways in Shewanella loihica Strain PV-4

Shewanella loihica strain PV-4 harbors both a functional denitrification (NO3 (-)→N2) and a respiratory ammonification (NO3 (-)→NH4 (+)) pathway. Batch and chemostat experiments revealed that NO2 (-) affects pathway selection and the formation of reduced products. Strain PV-4 cells grown with NO2 (-) as the sole electron acceptor produced exclusively NH4 (+). With NO3 (-) as the electron acceptor, denitrification predominated and N2O accounted for ∼90% of reduced products in the presence of acetylene. Chemostat experiments demonstrated that the NO2 (-):NO3 (-) ratio affected the distribution of reduced products, and respiratory ammonification dominated at high NO2 (-):NO3 (-) ratios, whereas low NO2 (-):NO3 (-) ratios favored denitrification. The NO2 (-):NO3 (-) ratios affected nirK transcript abundance, a measure of denitrification activity, in the chemostat experiments, and cells grown at a NO2 (-):NO3 (-) ratio of 3 had ∼37-fold fewer nirK transcripts per cell than cells grown with NO3 (-) as the sole electron acceptor. In contrast, the transcription of nrfA, implicated in NO2 (-)-to-NH4 (+) reduction, remained statistically unchanged under continuous cultivation conditions at NO2 (-):NO3 (-) ratios below 3. At NO2 (-):NO3 (-) ratios above 3, both nirK and nrfA transcript numbers decreased and the chemostat culture washed out, presumably due to NO2 (-) toxicity. These findings implicate NO2 (-) as a relevant modulator of NO3 (-) fate in S. loihica strain PV-4, and, by extension, suggest that NO2 (-) is a relevant determinant for N retention (i.e., ammonification) versus N loss and greenhouse gas emission (i.e., denitrification).