Purification and Conformational Analysis of a Key Exopolysaccharide Component of Mixed Culture Aerobic Sludge Granules

The application of aerobic sludge granules in wastewater treatment could increase the intensity of wastewater treatment processes because of their greater density and size relative to conventional sludge flocs. It has been suggested that granules are distinguished from flocs by gel forming exopolysaccharides. In this study, evidence is presented linking a specific exopolysaccharide component with granule extracellular polymeric substance (EPS) gelation. Granular EPS comprised three components: high-molecular-weight (MW) exopolysaccharide, medium-MW proteins and glycosides, and low-MW proteins and glycosides. The high-MW fraction was separated by fractional precipitation and preparatory-scale gel permeation chromatography (GPC). The MW profile of this fraction appears to be exclusively attributable to high-MW polysaccharide. The exopolysaccharide exists as a gel at normal wastewater treatment operating pH (i.e., 6.0-8.5), whereas the low/medium-MW material does not. Conformational analysis by atomic force microscopy (AFM) of the dried material showed that the polysaccharide forms pearl-necklace-like, intramolecularly condensed structures when dissolved in Milli-Q water and partially relaxed helical aggregates when in alkali solution. Consequently, the gel-forming property of EPS in the aerobic sludge granules tested is probably associated with high-MW polysaccharide components.