Acyl-Homoserine Lactone Enhances the Resistance of Anammox Consortia under Heavy Metal Stress: Quorum Sensing Regulatory Mechanism

Anaerobic ammonium oxidation (anammox) represents an energy-efficient process for the removal of biological nitrogen from ammonium-rich wastewater. However, the susceptibility of anammox bacteria to coexisting heavy metals considerably restricts their use in engineering practices. Here, we report that acyl-homoserine lactone (AHL), a signaling molecule that mediates quorum sensing (QS), significantly enhances the nitrogen removal rate by 24% under Cu²⁺ stress. A suite of macro-/microanalytical and bioinformatic analyses was exploited to unravel the underlying mechanisms of AHL-induced Cu²⁺ resistance. Macro-/microanalytical evidence indicated that AHL regulations on the production, spatial distribution, and functional groups of extracellular polymeric substances were not significant, ruling out extracellular partitioning and complexation as a principal mechanism. Meanwhile, molecular biological evidence showed that AHL upregulated the transcriptional levels of resistance genes (<i>sod</i>, <i>kat</i>, <i>cysQ</i>, and <i>czcC</i> responsible for antioxidation defense, Cu²⁺ sequestration, and transport) to appreciable extents, indicating intracellular resistance as the primary mechanism. This study yielded a mechanistic understanding of the regulatory roles of AHL in extracellular and intracellular resistance of anammox consortia, providing a fundamental basis for utilizing QS regulation for efficient nitrogen removal in wastewaters with heavy metal stress.