The bacterium Pseudomonas aeruginosa negatively regulates denitrification under anerobic conditions by two acyl-homoserine lactone quorum-sensing (QS) systems called las and rhl. However, it is unknown whether these systems have the same effect on denitrification in aerobic conditions. In this study, we investigated the regulation of las and rhl systems on aerobic denitrification. We showed that the removal of nitrate in P. aeruginosa PAO1 was repressed by both the las and rhl systems. The las and rhl systems had negative effects on activities of denitrifying enzymes NAP, NIR, NOR, and NOS. At the level of transcription, both QS systems inhibited the expression of target genes napA, nirS, norB, norC, and nosZ. Furthermore, the addition of an acylase, which degrades the acyl-homoserine lactone signals (AHLs), to wild type resulted in an increase in the removal of nitrate. Additionally, in aerobic denitrification process, the transcription factor DNR, which controls denitrification, was repressed by both QS systems. The results implied that modulation of QS in denitrifying bacteria, possibly through quorum quenching or QS inhibition, could help to improve the reduction of nitrate in wastewater treatment.