The antibacterial activity of hydroxylated fatty acids produced by Psuedomonas aeruginosa PR3 using ricinoleic acid, eicosadienoic acid and conjugated linoleic acid as substrates was investigated. The hydroxylated fatty acids showed different antibacterial activity against a range of food-borne pathogenic bacteria. The bioconverted ricinoleic acid exhibited the strong antibacterial activity against four Gram-positive bacteria, Bacillus subtilis ATCC 6633, Listeria monocytogenes ATCC 19166, Staphylococcus aureus ATCC 6538 and S. aureus KCTC 1916 and one Gram-negative bacteria, P. aeruginosa KCTC 2004. The bioconverted conjugated linoleic acid was slightly susceptible against 2 Gram-positive bacteria, B. subtilis ATCC 6633 and S. aureus KCTC 1916 and the bioconverted eicosadienoic acid against 4 Gram-positive bacteria, B. subtilis ATCC 6633, L. monocytogenes ATCC 19166, S. aureus ATCC 6538 and S. aureus KCTC 1916.