Abstract

When denitrifying bacteria such as Paracoccus denitrificans respire anaerobically they convert nitrate to dinitrogen gas via a pathway which includes the potent greenhouse gas, nitrous oxide (N2O). The copper-dependent enzyme Nitrous Oxide reductase (Nos) catalyzes the reduction of N2O to dinitrogen. In low-copper conditions, recent experiments in chemostats have demonstrated that Nos efficiency decreases resulting in significant N2O emissions. For the first time, a chemostat-based mathematical model is developed that describes the anaerobic denitrification pathway based on Michaelis–Menten kinetics and published kinetic parameters. The model predicts steady-state enzyme levels from experimental data. For low copper concentrations, the predicted Nos level is significantly reduced, whereas the levels for the non copper-dependent reductases in the pathway remain relatively unaffected. The model provides time courses for the pathway metabolites that accurately reflect previously published experimental data. In the absence of experimental data purely predictive analyses can also be readily performed by calculating the relative Nos level directly from the copper concentration. Here, the model quantitatively estimates the increasing level of emitted N2O as the copper level decreases.

Highlights

  • Nitrous oxide (N2O) is the third largest contributor to global warming behind carbon dioxide (CO2) and methane

  • In order to select the mathematical form of the dependence between the copper level and the Nitrous Oxide reductase (Nos) level, we look at N2O production rates

  • The model is calibrated from the nitrate-sufficient carbon-limited P. denitrificans results of (Felgate et al 2012) because they result in the most significant emissions of N2O

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Summary

Introduction

Nitrous oxide (N2O) is the third largest contributor to global warming behind carbon dioxide (CO2) and methane. Specific enzymes in the denitrification pathway have been the subject of detailed biochemical studies (e.g., Field et al 2008) together with the effect exerted by genetic regulation (e.g., Bergaust et al 2012) This wealth of information regarding the enzymes, and their kinetic behavior, has yet to be integrated into a robust mathematical model of the chemical reactions. Where reactions (1)–(4) are catalyzed by Nitrate reductase (Nar), Nitrite reductase (Nir), Nitric Oxide reductase (Nor), and Nitrous Oxide reductase (Nos), respectively These reactions consume a total of 10 electrons and 12 protons when two NOÀ3 ions are converted to a single molecule of N2 gas. By seeking the steady-state solution, expressions that estimate the reductase concentrations are obtained Using these values, time courses for NOÀ3 and other relevant metabolites can be calculated. For a given experiment the model provides a method of predicting the enzyme concentrations and the time courses for the metabolites. The model is available for download from http://www. uea.ac.uk/computing/software/modelling-denitrification

Experimental Procedures
Results
Discussion

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