The current study prepared, characterized, and evaluated a bitter apple seed ethanolic extract (BASEE) for its antioxidant, anticancer, antibacterial, and antifungal properties, as well as its use in preserving minced beef. The bitter apple seed ethanolic extract had a total phenolic content of 32 mg GAE g^-1 extract. BASEE contains dominant compounds, including ellagic (0.9 mg g^-1), cinnamic (0.8 mg g^-1), or gallic (0.6 mg g^-1) acids. The bitter apple seed ethanolic extract exhibited antioxidant activity by inhibiting DPPH free radicals. It exhibited activity towards Staphylococcus aureus and Escherichia coli, with effectiveness depending on the concentration. BASEE (400 μg mL^-1) significantly inhibited F. oxysporum mycelial growth, resulting in a 94% reduction in development. HCT116 cells treated with BASEE for 48 h stopped their growth, with IC₅₀ values of 55 μg mL^-1 for HCT116 cells and 100 μg mL^-1 for A549 cells. However, IC₅₀ values were significantly higher than those of Vero normal cells (1000 μg mL^-1). Results indicate that HCT116 cells showed greater sensitivity to the bitter apple seed ethanolic extract compared to the A549 cells and normal cells (Vero). The findings indicate that BASEE may function as an alternative antioxidant, antibacterial, antifungal, and anticancer agent. Bitter apple seed ethanolic extract can be effectively used as a natural preservative and a better alternative to synthetic compounds.