Identification of partial denitrification granulation enhanced by low C/N ratio in the aspect of metabolomics and quorum sensing

Partial denitrification granular sludge (PDGS) and denitrification granular sludge (DGS) play an important role in nitrogen removal from wastewater. However, the inherent cause of aggregation capacity related to the ratio of COD to nitrogen (COD/N) is still unclear. In this study, metabolomics analysis was combined with microbiological analyses, granular performance and extracellular polymeric substances (EPS) structure to explore the granulation mechanism at different influent COD/N ratios. The results showed that the higher COD/N ratio selectively enhanced the gluconeogenesis pathway, purine and pyrimidine metabolism pathway, resulting in more extracellular polysaccharide (PS) excretion and floc sludge. The absence of carbon source weakened tricarboxylic acid cycle (TCA) reaction, resulting in NAD+ and ADP decrease, nitrite accumulation and change of microbial community structure. The amino acids biosynthesis pathway was enhanced under low COD/N ratio, which promoted the hydrophobicity of EPS. PDGS had stronger Acyl-homoserine lactones (AHLs)-based quorum sensing (QS) than DGS during the operational period. CO8-HSL, C8-HSL and C6-HSL, as the main form of AHLs, played a dominating role in DGS and PDGS. Batch tests illustrated that adding AHLs obviously improved the synthesis of the amino acids, threonine (Thr), tryptophan (Trp), methionine (Met) and glycine (Gly). Dosing AHLs regulated PS synthesis only at a high COD/N ratio. The glucose-6P, glycerate-3p and UDP-Glc were up-regulated only in DSG, which increased the hydrophilic groups in EPS. The results not only provided the new insights into the metabolism of denitrifying granular sludge, but also indicated the application potential of the technologies regarding start-up and operation of granule sludge.