Bacillus subtilis KU201 was isolated from kimchi and characterized for probiotic use. B. subtilis KU201 was stable in artificial gastric condition, and adhered strongly against intestinal cell. B. subtilis KU201 was not produce a carcinogenic enzyme. B. subtilis KU201 showed a broad-spectrum antimicrobial effect. Bacteriocin KU201 was sensitive against protease XIV, proteinase K, and ${alpha}$-chymotrypsin, stable over a pH range of 3 to 9 for 4 h, and withstood exposure to temperatures of $50-90^{circ}C$ for 30 min. The molecular weight of bacteriocin KU201 was 3.5 kDa. The bactericidal effect of bacteriocin KU201 was observed by morphological changes in cell membrane. These results show that B. subtilis KU201 may be used as a potential probiotic strain, also bacteriocin KU201 can be used as a natural food preservative.