An antimicrobial peptide produced by Bacillus subtilis KU25, KU43, and KU44 was isolated from traditional Korean fermented foods and characterized. It was named as subtilin KU25, KU43, and KU44. Subtilin KU25, KU43, and KU44 were sensitive against ${alpha}$-chymotrypsin, protease XIII, and various proteinase enzymes, respectively. B. subtilis KU43 was selected as the producer with the broadest antimicrobial spectrum. Subtilin KU43 was stable at a pH range of 3 to 9 for 4 h, and withstood exposure to temperatures of $50-90^{circ}C$ for 30 min. The mode of inhibition against Listeria monocytogenes ATCC 15313 involved a bactericidal effect by a reduction in the cell numbers and breakage of the indicator cell membranes. The molecular mass of subtilin KU43 was measured at approximately 3.5 kDa. These results demonstrate the development of novel strains from traditional Korean fermented foods, and illustrate the possibility that some of these strains might generate a natural preservative compound.