Enterotoxins produced by Staphylococcus aureus are the key pathogenicity factors that can cause a variety of illnesses in humans, including staphylococcal gastroenteritis and food poisoning. It has been proven that licochalcone A is a potentially effective antimicrobial agent against S. aureus. In this study, Western blot assays, tumour necrosis factor release assays, murine T-cell proliferation assays, and real-time reverse transcriptase-PCR were performed to evaluate the effect of subinhibitory concentrations of licochalcone A on the secretion of two major enterotoxins (SEA and SEB) by S. aureus. The results show that licochalcone A significantly decreased, in a dose-dependent manner, the secretion of SEA and SEB by both methicillin-sensitive S. aureus and methicillin-resistant S. aureus. These results may increase the desirability of using licochalcone A as a lead compound for the design of more potent antibacterial agents based on the chalcone template.