This study determined the preservation effects of bacteriocin XJS01 on raw beef during storage at 4 degrees C. An integrated time- and concentration-dependent metabolomic analysis revealed that XJS01 preserved beef by influencing metabolic pathways of spoilage-induced bacteria. These pathways were primarily associated with meat spoilage, cell structure and stress response. The results also demonstrated that the antibacterial activity of XJS01 contributed to preservation by indirectly suppressing the formation of dipeptides. Additionally, 37 metabolites were significantly correlated with meat-quality traits, which were primarily related to valine, leucine and isoleucine degradation and alpha-linolenic acid metabolism. Meanwhile, 1,2,10-trihydroxydihydro-translinalyl oxide7-O-beta-D-glucopyranoside and 1-(beta-D-ribofuranosyl)-1,4-dihydronicotinamide were identified as potential biomarkers for estimating beef storage time under XJS01 protection. These findings provide valuable insights into the potential applications of bacteriocins for enhancing preservation and quality assessment in meat.