In this study we isolated and identified the lipopeptides produced by Bacillus amyloliquefaciens strains 1/6k and 4/9, which were isolated from Adriatic Sea. Lipopeptides belong to the family of cyclic biosurfactants representing a group of surface active compounds produced by various microorganisms. Studied Bacillus spp. strains were able to produce cyclic lipopeptides surfactin, iturin A and fengycin. The characterisation of lipopeptides was studied by various analytical techniques (high-performance liquid chromatography – HPLC and matrix-assisted laser desorption/ionization time-of-flight analysis for molecular mass determination – MALDI-TOF/MS). These methods confirmed co-production of three lipopeptides – surfactin, iturin A and fengycin by Bacillus amyloliquefaciens.