The aim of this study was to identify probiotic possibility of lactic acid bacteria (LAB) from mustard leaf kimchi and investigate synergy effects of combination with LAB and fructooligosaccharide (FOS) on antimicrobial activity. Lactobacillus acidophilus GK20, L. paracasei GK74, and L. plantarum GK81 have high survival ratio under gastric and intestinal fluids and were resistant to ampicillin, streptomycin, or tetracycline. The pH of LAB gradually decreased but the titratable acidity increased owing to organic acids production. Especially, L. acidophilus GK20 and L. paracasei GK74 produced hydrogen peroxide and bacteriocin. When LAB was co-cultured with Listeria monocytogenes or Staphylococcus aureus in broth without FOS, the inhibition ratio of L. monocytogenes and S. aureus by L. acidophilus GK20 was 36 and 57%, but L. paracasei GK74 showed higher inhibitory effect against L. monocytogenes than S. aureus. Meanwhile, the cell growth, culture pH, titratable acidity, and bacteriocin activity of L. acidophilus GK20 and L. paracasei GK74 were significantly (p<0.05) increased in MRS with FOS than without prebiotic. In conclusion, L. acidophilus GK20 and L. paracasei GK74 could be potentially used as novel probiotic strains, moreover, the combination between L. acidophilus GK20 or L. paracasei GK74 and FOS has great potential as a synbiotic.