Insights into simultaneous anammox and denitrification system with short-term pyridine exposure: Process capability,inhibition kinetics and metabolic pathways

• Short-term effect of the pyridine exposure on the SAD process was investigated. • The SAA at 150 mg/L pyridine reduced by 56.7% of the maximum value. • Inhibition kinetics models and inhibitory parameters were indicated. • Collaboration of AnAOB, HDB and PDB promoted the SAD. • Possible metabolic pathways of nitrogen and pyridine were proposed. In-depth knowledge on the role of pyridine as a bottleneck restricting the successful application of anammox-based process treating refractory coking wastewater remains unknown. In this study, the effect of short-term pyridine addition on a simultaneous anammox and denitrification (SAD) system fed with 25–150 mg/L pyridine was explored. The short-term operation showed that the highest total nitrogen (TN) removal efficiency was achieved at 25–50 mg/L of pyridine. As the pyridine addition increased, the contribution of the anammox pathway in nitrogen removal decreased from 99.3% to 79.1%, while the denitrification capability gradually improved. The specific anammox activity (SAA) at 150 mg/L pyridine decreased by 56.7% of the maximum SAA. The modified non-competitive inhibition model indicated that the 50% inhibitory concentration (IC₅₀) of pyridine on anammox was 84.18 mg/L and the substrate inhibition constant (K_i) of pyridine for self-degradation was 135.19 mg/L according to the Haldane model. Moreover, high-throughput sequencing confirmed the abundance of Candidatus Kuenenia as the amount of anammox species decreased, while the amounts of denitrifiers and pyridine degraders significantly increased as the pyridine stress increased. Finally, the possible pathways of nitrogen bioconversion and pyridine biodegradation in the SAD system were elucidated through metagenomic analysis and gas chromatography/mass spectrometry results. The findings of this study enlarge the understanding of the removal mechanisms of complex nitrogenous pyridine-containing wastewater treated by the SAD process.