Enhancement of aerobic sludge granulation by quorum sensing signaling molecules mediated by biomimetic bacterial extracellular vesicles

The addition of exogenous quorum sensing signaling molecules to the activated sludge system enables rapid sludge granulation. However, signaling molecules exposed to the environment are easily degraded, and their quorum sensing effects cannot be maintained in the long term. Therefore, they must be frequently added, which leads to an increase in operational costs. In this study, signaling molecules, acylated homoserine lactones (AHLs), were packaged into biomimetic bacterial extracellular vesicles-lipid vesicles (AHLs@ZZT), and the mediating role of lipid vesicles for AHLs was utilized to improve the utilization efficiency of signaling molecules. The results show that packaging of AHLs into vesicles slowed down the release of AHLs and protected them from degradation by porcine renal acylase. Moreover, the chemical properties of lipid vesicles are similar to the structure of bacterial cell walls, making it easier for the packaged AHLs to bind to target bacteria, thereby generating more EPS and accelerating the aggregation of microorganisms. After adding the lipid vesicles loaded with AHLs into the activated sludge system, the sludge granulation process was accelerated. Furthermore, the frequency of AHLs@ZZT addition was reduced by three times compared to that of unpackaged AHLs, resulting in a granular sludge with a smoother and denser surface. High-throughput sequencing of the sludge genome in the system indicated that adding AHLs@ZZT could promote microbial diversity and enrich functional microorganisms. Three-dimensional fluorescence spectra further revealed that the tryptophan and tyrosine proteins in extracellular polymeric substances secreted by these microorganisms facilitated sludge granulation. In summary, the lipid vesicle-mediated signaling molecular strategy can promote the rapid formation of granular sludge and maintain the long-term stability of the granular sludge structure, representing an economic and feasible regulation strategy.