Microbial spoilage poses challenges to food safety, prompting the development of a redox-driven photodynamic inactivation (PDI) system using vitamin C (VC) and vitamin B2 (VB2) to enhance bactericidal efficiency. This innovation selectively increases hydrogen peroxide (H2O2) production, leading to highly cytotoxic hydroxyl radicals (OH) generation and >99.999% eradication of Pseudomonas fragi. Mechanistic analyses reveal a two-stage bactericidal mechanism disrupting bacterial metabolism and membrane integrity. The system extends beef shelf life by 2 days through delaying microbial growth and preserving physicochemical properties. This non-thermal strategy offers promise in food preservation, emphasizing the importance of redox-mediated antibacterial mechanisms.