Abstract: This study focused on fresh-keeping effect of antimicrobial peptides from Bacillus methylotrophilus in Tilapia fillet preservation. The total plate count, sensory quality, physical and chemical indicators of Tilapia fillets which were soaked in antimicrobial peptides from Bacillus methylotrophilus were detected in the experiment. The results showed that the shelf life of the samples test group could be prolonged at least 4d and the optimum concentration was 1.5 to 2.0 percent. As a biological preservative of chilled meat the antimicrobial peptides from Bacillus methylotrophilus F35 had broad application prospects.