In order to study the antibacterial properties of antibacterial peptide Brevilaterin secreted by Brevibacillus laterosporus, effect of different external factors of food on its antibacterial activity were investigated, and its MIC and MBC values in varied internal environment of food were further checked. The results showed that Brevilaterin exhibited good stability towards those selected metal ions, emulsifiers (except soybean phospholipid), thickeners, enzymes, high concentration of sugar and repeated treatment of freeze-thaw. Furthermore, MIC and MBC values of Brevilaterin against S. aureus, L. monocytogenes, M. luteus, P. aeruginosa, L. lactis and H. alkalicola could be significantly changed with variation of the temperature, pH and osmotic pressure of food. Compared with the effect at the optimal temperature (37℃) of those foodborne pathogens mentioned above, the MIC and MBC values of Brevilaterin on M. luteus decreased to 1/8 at 4℃, the MIC and MBC values of Brevilaterin on P. aeruginosa, L. monocytogenes and S. aureus were decreased to 1/4~1/2 at 10℃. Compared with the effect when strains were grown at the optimum pH (6~7), the MIC and MBC of Brevilaterin on L. lactis were reduced to 1/2 at pH 2, and the MIC and MBC of Brevilaterin on S. aureus were reduced to 1/2 at pH 8~9. Similarly, the MIC and MBC of Brevilaterin for H. kalicola at pH 7 were reduced to 1/4 of that at pH 11 (optimal pH). The MIC and MBC of Brevilaterin for S.aureus under hypertonic condition were reduced to 1/2 of that under normal pressure. Therefore, when the growth conditions of foodborne pathogens deviated from their optimal conditions, Brevilaterin showed stronger antibacterial effects, while both of MIC and MBC values were reduced to 1/8~1/2 of that at optimal conditions. All of these properties exhibited by Brevilaterin indicated its scientific guideline on the food application in the future.