ABSTRACTThe threat of antibiotic resistant bacteria has called for alternative antimicrobial strategies that would mitigate the increase of classical resistance mechanism. Many bacteria employ quorum sensing (QS) to govern the production of virulence genes and formation of drug-resistance biofilms. Blocking QS mechanisms have proven to be a functional alternative to conventional antibiotic control of infections. The concepts of quorum sensing inhibitors (QSI) and quorum quenching enzymes (QQ) have been investigated separately. In this study however, we simulated the synergistic effect of QQ and QSI in blocking bacterial QS. This effect was validated by experiments using AiiA and G1 as QQ and QSI respectively onPseudomonas aeruginosaLasR/I and RhlR/I QS circuits. The combination of a QQ and a QSI almost completely blocked theP. aeruginosaQSlasandrhlsystem. Our findings provided a potential application strategy for bacterial QS disruption.