Abstract The aim of this study was to investigate the biological inhibiting effect of a beef‐derived Latilactobacillus sakei (RS‐25) on the spoilage of beef steaks in overwrapped packaging during the 12 days of storage at 4°C. Beef quality as well as microbial indicators were determined at different intervals during the storage after the inoculation of RS‐25 at the 6 log CFU/g, and the high‐throughput sequencing was applied to investigate the changes of microbial community structure during the storage. The inoculation of RS‐25 on beef had no effect (p > .05) on pH, TBARS, and TVB‐N during storage indicating the weak effect of such strain on the eat quality. Furthermore, the rise of L* and the delayed decline of a* and b* reveal the protection effect of RS‐25 on the meat color. RS‐25 reduced the re‐contaminated Salmonella typhimurium by 1.16 log CFU/g (p
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