Abstract

In this study, activated sludge (AS) presented high aggregation ability of 56.9% to form flocs. To elucidate the effects of extracellular polymeric substances (EPS) on the AS flocs formation, four EPS fractions, including soluble EPS (S-EPS), loosely bound EPS (LB-EPS), ionic tightly bound EPS (ionic TB-EPS) and hydrophobic tightly bound EPS (hydrophobic TB-EPS), were extracted and characterized. The ionic TB-EPS were the largest fraction, accounting for 51.3% of the total EPS. By extracting EPS fractions, it was shown that ionic TB-EPS followed by hydrophobic TB-EPS played positive contribution to the formation of AS flocs, while both S-EPS and LB-EPS had negative effects. Furthermore, chemical reagents (Urea, SDS and EDTA) assays, zeta potential and spectroscopic assays were conducted to explore the role of hydrogen bonds, electrostatic interaction and hydrophobic interaction in the formation of AS flocs. The result showed that ionic TB-EPS contained sufficient cation-binding sites, such as -OH and CO-O- groups, and promoted AS aggregation through ion bridging. Hydrophobic TB-EPS contained hydrophobic groups and promoted AS aggregation through hydrophobic action. A high proportion of uronic acids resulted in high surface negative charge, which might be the reason that LB-EPS and S-EPS inhibited AS aggregation through electrostatic repulsion. From the perspective of molecule structure, the proteins in ionic TB-EPS had a loose structure, which fully exposed the inner cation-binding sites. This might be the critical reason why ion bridging acted as the main force for the formation of AS flocs. This study provided a new sight for the extraction of EPS fractions and expanded the understanding of EPS in the formation of AS flocs.

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