Abstract: The present disclosure discloses Lactobacillus sakei ZFM225, with a preservation number of CCTCC NO:M 2016669. The present disclosure further discloses an isolation and purification method of an Lactobacillus sakei ZFM225 bacteriocin, including the steps of: precipitating fermentation supernatant of the Lactobacillus sakei ZFM225 by an ammonium sulfate precipitation method to obtain crude protein; desalting the crude protein to obtain crude protein extract; isolating the crude protein extract by cation exchange chromatography, performing elution with a gradient eluent containing 0.5 M to 1.0 M NaCl, and desalting the obtained eluate to obtain a preliminary protein purified solution; and then purifying the preliminary protein purified solution by reversed-phase high-performance liquid chromatography to obtain the Lactobacillus sakei ZFM225 bacteriocin. The Lactobacillus sakei ZFM225 bacteriocin has antibacterial activity on both gram-positive bacteria and gram-negative bacteria.