A quantified nitrogen metabolic network by reaction kinetics and mathematical model in a single-stage microaerobic system treating low COD/TN wastewater

A single-stage intermittent aeration microaerobic reactor (IAMR) has been developed for the cost-effective nitrogen removal from piggery wastewater with a low ratio of chemical oxygen demand (COD) to total nitrogen (TN). In this study, a quantified nitrogen metabolic network was constructed based on the metagenomics, reaction kinetics and mathematical model to provide a revealing insight into the nitrogen removal mechanism in the IAMR. Metagenomics revealed that a complex nitrogen metabolic network, including aerobic ammonia and nitrite oxidation, anammox, denitrification via nitrate and nitrite, and nitrate respiration, existed in the IAMR. A novel method for solving kinetic parameters with high stability was developed based on a genetic algorithm. Use this method to calculate the kinetics of various reactions involved in nitrogen metabolism. Kinetics revealed that simultaneous partial nitritation-anammox (PN/A) and partial denitrification-anammox (PDN/A) were the dominant approaches to nitrogen removal in the IAMR. Finally, a kinetics-based model was proposed for quantitatively describing the nitrogen metabolic network under the limitation of COD. 58% ∼ 67% of nitrogen was removed via the anammox-based processes (PN/A and PDN/A), but only 7% ∼ 12% and 1% ∼ 2% of nitrogen were removed via heterotrophic denitrification of nitrite and nitrate, respectively. The half-inhibition constant of dissolved oxygen (DO) on anammox was simulated as 0.37 ∼ 0.60 mg L⁻¹, filling the gap in quantifying DO inhibition on anammox. High-frequency intermittent aeration was identified as the crucial measure to suppress nitrite-oxidizing bacteria, although it has a high affinity for DO and NO₂⁻-N. In continuous aeration mode, the simulated NO₃⁻-N in the IAMR would rise by 39.6%. The research provides a novel insight into the nitrogen removal mechanism in single-stage microaerobic systems and provides a reliable approach to practicing PN/A and PDN/A for cost-effective nitrogen removal.