This study was carried out to get basic resources for the industrial use of Lactobacillus bulgaricus SP5. The antibacterial activity of the supernatant obtained from Lactobacillus bulgaricus SP5 was tested against the pathogenic bacteria such as Escherichia coli KCCM 11234, Salmonella enteritidis KCCM 3313, Salmonella enteritidis KCCM 12021, and Salmonella typhimurium KCCM 40253. The supernatant of L. bulgaricus SP5 showed antibacterial activity against tested pathogenic bacteria. The antibacterial activity was examined after adjusting pH and heat treatment of supernatant. Heat treatment of supernatant had antibacterial activity against pathogenic bacteria at all temperature. However, pH changes showed no antibacterial activity. Antibacterial activity of the supernatant was confirmed to be due to organic acids (lactic, acetic, phosphoric, succinic, pyroglutamic, citric, malic, and formic acid).