To develop a lactic starter to produce antimicrobial substance for inhibiting the growth of a variety of foodborne spoilage bacteria in fermented foods, we investigated the antibacterial effect of the antibacterial substance, produced by Lactobacillus amylovorus IMC-1, against foodborne spoilage strains, and its sensitivity on the treatment of proteolytic enzymes. L. amylovorus IMC-1, which was isolated from a traditional cheese in Inner Mongolia, produced a maximum amount of antibacterial substance in the skim milk medium after 72 h incubation at 37℃, and further incubation resulted in the same activity. The substance obtained from gel filtration inhibited all strains used such as Bacillus subtilis IFO 3025, Staphylococcus aureus IAM 1011, Listeria monocytogenes VTU 206, Escherichia coli RB, and Pseudomonas fragi IFO 3458 at the concentration of 20 units/ml. This substance was found to show bactericidal action against B. subtilis, E. coli, and Ps. fragi, and bacteriostatic activity against both Staph. aureus and L. monocytogenes. The bactericidal action was due to cellular lysis. The substance is not organic acid, hydrogen peroxide and proteinaceous compound.