In recent years functional foods have become increasingly popular. Products which include viable probiotic culture (for example lactic acid bacteria) belong to functional foods. Lactic acid bacteria play a main role in the natural preservation of food products. This microorganisms produce several metabolic compounds with antagonistic properties against spoilage, pathogenic bacteria, yeasts and moulds. Production ability of antagonistic compounds depends on several factors among other things temperature and time incubation. The aim of this study was to determine of growth conditions (temperature, time) of Lactobacillus plantarum strains and to determine the antagonistic properties against gram-positive and gram-negative bacteria. All the chosen Lactobacillus plantarum strains are able to inhibit used indicator bacterial strains. The influence of the time and temperature incubation lactic acid bacteria on antagonistic activity against indicator bacteria was observed. The biggest inhibition zones were obtained applying incubation time 48 hours at temperature 37ºC. The Lactobacillus plantarum strains, which were incubated at 6ºC did not inhibit growth of E. coli ATCC 25922, P. mirabilis 180, K. ornithinolytica.