Characterization of EPS subfractions from a mixed culture predominated by partial-denitrification functional bacteria

Extracellular polymeric substances (EPS) play a crucial role in the aggregation of partial denitrification (PD) consortia, as EPS is closely linked to bioreactor performance. However, the structural and compositional properties of EPS from PD consortia have not yet been investigated. In this study, photometric measurements indicated that PD consortia contained significantly more EPS (168.81 ± 2.10 mg/g VSS) compared to conventional activated sludge (79.79 mg/g VSS). The EPS of PD consortia exhibited a significant predominance of proteins over polysaccharides, with a protein/polysaccharide ratio of 1.43 ± 0.10. FTIR analysis revealed that the EPS of PD consortia contained fewer hydrophilic functional groups, particularly carboxyl and carbonyl groups, indicating a high aggregation potential. The content comparison of EPS and functional groups across three stratified EPS subfractions from PD consortia consistently followed the sequence: TB-EPS > LB-EPS > S-EPS. XPS results corroborated the FTIR findings and the protein/polysaccharide ratio determined by photometric measurements, all of which suggested that the EPS of PD consortia exhibited a higher abundance of hydrophobic functional groups. However, the higher α-helix/(β-sheet + random coil) ratio (0.99) suggested that the proteins in PD consortia had a compact structure, making inner hydrophobic groups difficult to expose. This compact protein structure could limit aggregation among bacterial cells, indicating the need for process optimization to enhance sludge aggregation in PD-related processes. Overall, understanding the aggregation characteristics of PD consortia could improve the application of PD-based processes.