Cyclic lipopeptides were produced by Bacillus amyloliquefaciens subsp. plantarum strain BC32-1 that was isolated from yellow loess soil in the Jeonnam province of South Korea. Several lipopeptides were isolated from the bacteria using organic solvent extraction and reverse-phase high-performance liquid chromatography (RP-HPLC). Purified iturin-, surfactin-, and fengycin-type lipopeptides were identified using liquid chromatographymass spectrometry (LC-MS) analysis. Among the lipopeptides, C 17 -fengycin B showed strong antifungal activity against the phytopathogenic fungus, Fusarium oxysporum f. sp. radicis-lycopersici, and then the fengycin was further characterized by UV, Fourier transform-infrared spectroscopy (FT-IR), and LC-MS/MS analyses. C 17 -fengycin B was highly produced at quantities of up to 15 µg/mL at 37 o C, whereas little amount of the fengycin was produced at 25 o C. Purified C 17 -fengycin B inhibited mycelial growth of F. oxysporum with a minimal inhibitory concentration of 50 µg/mL. This study suggests that C 17 -fengycin B is a major antifungal component produced by the BC32-1 strain that could be used as an environmentally friendly agent to control the phytopathogenic F. oxysporum.