Antibiotics are widely found in nitrogen-containing wastewater, which may affect the operation stability of anaerobic ammonium oxidation (anammox)-based biological treatment systems. Extracellular polymeric substances (EPSs) of anammox sludge play a pivotal role in combining with antibiotics; however, the exact role and how the structure of the leading component of EPSs (i.e., extracellular proteins) changes under antibiotic stress remain to be elucidated. Here, the interaction between sulfamethoxazole and the extracellular proteins of anammox sludge was investigated via multiple spectra and molecular simulation. Results showed that sulfamethoxazole statically quenched the fluorescent components of EPSs, and the quenching constant of the aromatic proteins was the largest, with a value of 1.73 × 104 M–1. The overall binding was an enthalpy-driven process, with ΔH = −75.15 kJ mol–1, ΔS = −0.175 kJ mol–1 K–1, and ΔG = −21.10 kJ mol–1 at 35 °C. The O-P-O and CO groups responded first under the disturbance of sulfamethoxazole. Excessive sulfamethoxazole (20 mg L–1) would decrease the ratio of α-helix/(β-sheet + random coil) of extracellular proteins, resulting in a loose structure. Molecular docking and dynamic simulation revealed that extracellular proteins would provide abundant sites to bind with sulfamethoxazole, through hydrogen bond and Pi-Akyl hydrophobic interaction forces. Once sulfamethoxazole penetrates into the cell surface and combines with the transmembrane ammonium transport domain, it may inhibit the NH4 + transport. Our findings enhance the understanding on the interaction of extracellular proteins and sulfamethoxazole, which may be valuable for deciphering the response property of anammox sludge under the antibiotic stress.

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High-rate contact stabilization is promising to capture organic carbon from municipal wastewater; however, its engineering application is restricted due to its poor capability of removing soluble organic matter (particle size < 0.1 μm). In this work, the carbon capture efficiency and robustness of the coagulation-enhanced high-rate contactstabilized process for treating municipal wastewater with a varied content of soluble organic matter was comprehensively evaluated. Results showed that the high-rate contact stabilization process did not perform well in the removal and capture of influent organic substances with a high proportion of soluble components. Adding coagulants into the above process favored organic carbon capture, whose recovery efficiency could reach up to 60% even under the 90% influent soluble chemical oxygen demand condition. Mechanism analysis revealed that the hydrolyzed product of the coagulant would adsorb dissolved components to form colloid and subcolloid particles, which was favored for further capture by sludge. The fitting result of the neutral community model indicated that both deterministic environmental factors and random processes influence the succession of sludge community. This work implied that the coagulation-enhanced high-rate contact stabilization process is feasible to recover organic carbon from municipal wastewater, which may shed a promising light for sustainable carbon control and management.

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