Quorum sensing is a cell-to-cell communication process in bacterial kingdom, in which the expression of genetic information is controlled via the production and detection of “signal molecules” in a population density-dependent manner. Quorum sensing system has been discovered and shown to control virulence factors in several Gram-negative bacterial pathogens in aquaculture, including Vibrio harveyi, a harmful bacterium in shrimp. It has been shown that the ability to inactivate AHL signal molecules may be useful in controlling the AHL-mediated virulence in many Gram-negative pathogenic bacteria. AHL-lactonase, one of the three AHL-disrupting enzyme classes, attracts significant interest of researchers due to its high substrate specificity toward AHL signals. In this study, a recombinant and purified AHL-lactonase (AiiAN26.2 protein) was verified for its ability to inhibit bioluminescence of V. harveyi cultures. AiiAN26.2 protein at a concentration of 15 µg ml-1 could significantly attenuate the luminescence intensity of V. harveyi cultures over a 6-hour period. The findings of the study are important for future research as it was proven that recombinant AHL-lactonase can be used to control infectious diseases in aquaculture which are related to the AHL-mediated quorum sensing system.