Oxygen consumption in the marine bacterium Pseudomonas nautica predicted from ETS activity and bisubstrate enzyme kinetics

Abstract

The respiratory O2 consumption in aerobic bacterial cultures has been modeled from the time profiles of the in vitro activity of the respiratory electron transfer system (ETS), the bacterial protein and the concentration of the carbon source in the cultures. The model was based on the concept of bisubstrate kinetic control of the ETS throughout the exponential, steady-state and senescent phases of the cultures. In the exponential phase, the measured rates of O2 consumption and the in vitro ETS activity were closely coupled, but in the senescent phase, they were uncoupled. The in vitro ETS activity remained high even after the culture's carbon source was exhausted, while the O2 consumption fell to low levels. Based on the hypothesis that this uncoupling was caused by limitation of the intracellular ETS substrates (NADH and NADPH), a semi-empirical model incorporating a bisubstrate enzyme kinetics algorithm was formulated and fitted to the observations of the experiments. The model predicted the rate of O2 consumption throughout the different phases of the cultures with an r2 > 0.92 (n = 9, P < 0.001) using physiologically realistic Michaelis and dissociation constants. These results suggest that plankton respiration in the field could be assessed more accurately than before by measuring the intracellular ETS substrates (NADH and NADPH), in addition to ETS activity, in plankton.