Abstract The global spread of multidrug-resistant Staphylococcus aureus (MRSA) poses serious threats to public health and food safety. In this study, the antibacterial potential of the anthraquinone compound 13394-2, derived from soil actinomycetes, was systematically evaluated. In vitro, 13394-2 exhibited potent activity (MIC as low as 0.25 μg/mL), rapid bactericidal effects, biofilm clearance, and concentration-dependent killing.