The aim of this study is to produce biosurfactants by Yarrowia lipolytica, Saccharomyces cerevisiae and Rhodotorula glutinis in different growth mediums and to evaluate antimicrobial and anti adhesive potential of these biosurfactants. Inhibition effect of biosurfactants were tested on Escherichia colt, Staphylococcus aureus and Pseudomonas aeruginosa. This study achieved the biosurfactant production at different amounts with a low-cost fermentative medium (frying oil waste and corn maceration liquid) by yeast species. The biosurfactant production yield and inhibitory effect of these biosurfactants were found dependent on composition of growth medium, species of yeast and pathogenic bacteria. The most biomass yield was obtained by Y. lipolytica in medium containing food waste. All biosurfactants had a certain inhibitory activity against pathogens at concentrations of 5 mg/mL and 10 mg/mL. Additionally, biosurfactants (10 mg/mL) indicated antiadhesion effect against S. aureus and P. aeruginosa, except E. colt. Despite the several advantages of biosurfactants, there is a few reports related to their use on food products and food processing. Biosurfactants from this study have economically the potential to be used as antibacterial substances in food industry for the purpose of cleaning of solid surfaces, tool and equipment, process media etc.