Bacillus velezensis GH1‐13 was recently isolated and showed to promote plant growth and strong antagonistic effects against phytopathogenic fungi including Gibberella moniliformis, Fusarium fujikuroi, Bipolaris oryzae, and Colletotrichum gloeosporioides. In this study, the antifungal substances produced by the GH1‐13 strain were purified by ethyl acetate solvent partitioning and solid phase extraction using Sep‐Pak C18 cartridge. The pure antifungal substances were finally purified by preparative reversed phase‐high performance liquid chromatography. We investigated the structure of the purified antifungal compounds using liquid chromatography mass spectrometry and nuclear magnetic resonance spectroscopy. These compounds were identified as surfactin‐type cyclic lipopeptides having an ELLVDLL amino acid sequence with different lengths of lipid chains. The surfactin lipopeptides effectively inhibited the growth of fungal mycelium, suggesting that the GH1‐13 strain will be a potential biocontrol agent against plant pathogenic fungi.
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