Low Probability of Initiating nirS Transcription Explains Observed Gas Kinetics and Growth of Bacteria Switching from Aerobic Respiration to Denitrification

In response to impending anoxic conditions, denitrifying bacteria sustain respiratory metabolism by producing enzymes for reducing nitrogen oxyanions/-oxides (NOx) to N2 (denitrification). Since denitrifying bacteria are non-fermentative, the initial production of denitrification proteome depends on energy from aerobic respiration. Thus, if a cell fails to synthesise a minimum of denitrification proteome before O2 is completely exhausted, it will be unable to produce it later due to energy-limitation. Such entrapment in anoxia is recently claimed to be a major phenomenon in batch cultures of the model organism Paracoccus denitrificans on the basis of measured e−-flow rates to O2 and NOx. Here we constructed a dynamic model and explicitly simulated actual kinetics of recruitment of the cells to denitrification to directly and more accurately estimate the recruited fraction (). Transcription of nirS is pivotal for denitrification, for it triggers a cascade of events leading to the synthesis of a full-fledged denitrification proteome. The model is based on the hypothesis that nirS has a low probability (, h−1) of initial transcription, but once initiated, the transcription is greatly enhanced through positive feedback by NO, resulting in the recruitment of the transcribing cell to denitrification. We assume that the recruitment is initiated as [O2] falls below a critical threshold and terminates (assuming energy-limitation) as [O2] exhausts. With = 0.005 h−1, the model robustly simulates observed denitrification kinetics for a range of culture conditions. The resulting (fraction of the cells recruited to denitrification) falls within 0.038–0.161. In contrast, if the recruitment of the entire population is assumed, the simulated denitrification kinetics deviate grossly from those observed. The phenomenon can be understood as a ‘bet-hedging strategy’: switching to denitrification is a gain if anoxic spell lasts long but is a waste of energy if anoxia turns out to be a ‘false alarm’.Author Summary: In response to oxygen-limiting conditions, denitrifying bacteria produce a set of enzymes to convert / to N2 via NO and N2O. The process (denitrification) helps generate energy for survival and growth during anoxia. Denitrification is imperative for the nitrogen cycle and has far-reaching consequences including contribution to global warming and destruction of stratospheric ozone. Recent experiments provide circumstantial evidence for a previously unknown phenomenon in the model denitrifying bacterium Paracoccus denitrificans: as O2 depletes, only a marginal fraction of its population appears to switch to denitrification. We hypothesise that the low success rate is due to a) low probability for the cells to initiate the transcription of genes (nirS) encoding a key denitrification enzyme (NirS), and b) a limited time-window in which NirS must be produced. Based on this hypothesis, we constructed a dynamic model of denitrification in Pa. denitrificans. The simulation results show that, within the limited time available, a probability of 0.005 h−1 for each cell to initiate nirS transcription (resulting in the recruitment of 3.8–16.1% cells to denitrification) is sufficient to adequately simulate experimental data. The result challenges conventional outlook on the regulation of denitrification in general and that of Pa. denitrificans in particular.