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Bacteria are the main cause of spoilage for fish and fishery products. Through the inactivation of the quorum sensing (QS) system, quorum quenching (QQ) enzymes can block the synthesis of bacterial virulence factors and effectively inhibit bacteria-induced food spoilage. This study analyzed the changes of microbiota in red sea bream filets during refrigerated storage. The results showed a decrease in microbial diversity with storage time, with Aeromonas veronii becoming the dominant bacteria on day 4. A novel N -acyl homoserine lactones (AHL) acylase PF2571, from the screened QQ bacterium Pseudomonas fluorescens PF08, was identified and expressed in Escherichia coli to evaluate its QQ efficiency and effects on spoilage potential. Spoilage-related QS factors of A. veronii BY-8, including biofilm formation, motility, and protease, lipase, and alginate production, were inhibited by PF2571. Its inhibitory effect on red sea bream spoilage was demonstrated by the lower freshness indicators for PF2571 treated filets. Our study demonstrates the potential of the QQ enzyme for prolonging the shelf life of fish and fishery products. • The dynamics and composition of the microbial communities in red sea bream filets during storage at 4 °C were analyzed. • The quorum quenching enzyme PF2571 was identified and heterologous expressed. • The purified PF2571 enzyme showed good QS inhibitory activity against A. veronii BY-8. • The purified PF2571 enzyme effectively improved the quality of red sea bream filets during storage. [ABSTRACT FROM AUTHOR]