Correlation of Denitrification-Accepted Fraction of Electrons with NAD(P)H Fluorescence for Pseudomonas aeruginosa Performing Simultaneous Denitrification and Respiration at Extremely Low Dissolved Oxygen Conditions

In cystic fibrosis airway infection, Pseudomonas aeruginosa forms a microaerobic biofilm and undergoes significant physiological changes. It is important to understand the bacteriumʹs metabolism at microaerobic conditions. In this work, the culture properties and two indicators (the denitrification-accepted e- fraction and an NAD(P)H fluorescence fraction) for the cultureʹs "fractional approach" to a fully anaerobic denitrifying state were examined in continuous cultures with practically zero DO but different aeration rates. With decreasing aeration, specific OUR decreased while specific NAR and NIR increased and kept YATP/S relatively constant. P. aeruginosa thus appeared to effectively compensate for energy generation at microaerobic conditions with denitrification. At the studied dilution rate of 0.06 h^-1, the maximum specific OUR was 2.8 mmol O2/g cells-h and the Monod constant for DO, in the presence of nitrate, was extremely low (<0.001 mg/L). The cell yield YX/S increased significantly (from 0.24 to 0.34) with increasing aeration, attributed to a roughly opposite trend of YATP/X (ATP generation required for cell growth). As for the denitrification-accepted e- fraction and the fluorescence fraction, both decreased with increasing aeration as expected. The two fractions, however, were not directly proportional. The fluorescence fraction changed more rapidly than the e- fraction at very low aeration rates, whereas the opposite was true at higher aeration. The results demonstrated the feasibility of using online NAD(P)H fluorescence to monitor sensitive changes of cellular physiology and provided insights to the shift of e--accepting mechanisms of P. aeruginosa under microaerobic conditions.