Metabolomic insights into the inhibition mechanism of methyl N-methylanthranilate: A novel quorum sensing inhibitor and antibiofilm agent against Pseudomonas aeruginosa

The quorum sensing (QS) inhibition effect of methyl N-methylanthranilate (MMA) from Pericarpium Citri Reticulatae Chachiensis against foodborne pathogen Pseudomonas aeruginosa was reported for the first time. MMA effectively attenuated QS related virulence factors production and biofilm formation, while suppressed expression of a dozen of QS related genes. Untargeted LC-MS metabolomics revealed 108 significantly altered metabolites after MMA treatment. They indicated that MMA addition reduced the efficiency of TCA cycle and antioxidant systems, disturbed amino acid and nucleotide metabolism, increased unsaturated fatty acid and decreased peptidoglycan components, which might ultimately attenuate P. aeruginosa pathogenicity and restrain biofilm formation. Physiological characterization confirmed the compromised membrane integrity and increased intracellular oxidative stress after MMA treatment. Furthermore, metabolomics data implied that MMA inhibition on QS might exert through disrupting QS autoinducer PQS biosynthesis, which was supported by molecular docking. Our data indicated that MMA could be used as a novel QS inhibitor and anti-biofilm agent to improve food safety. It also provided new insight in the possible underlying inhibition mechanism of MMA and the response of P. aeruginosa to MMA.