Fermented foods of Korean temple are vegetable-based foods and their seasonings are limited to five pungent herbs. The aim of this study was to investigate cell-free supernatants (CFS) of lactic acid bacteria isolated from the fermented foods of Korean temples. Three selected CFS significantly inhibited the biofilm formation of Escherichia coli O157:H7 at 24 h, while one of the selected CFS markedly inhibited the biofilm formation at 72 h. However, only one CFS significantly attenuated the biofilm formation of Staphylococcus aureus at 24 h, but these CFS failed to inhibit the biofilm formation. Moreover, selected CFS effectively inhibited the growth of E. coli O157:H7, whereas the growth of S. aureus was not affected in the presence of CFS. These results suggest that CFS of lactic acid bacteria isolated from fermented foods of Korean temples shows antimicrobial activity agasint Gram-negative E. coli O157:H7, but not at Gramd-positive S. aureus .