| Summary: | In the past decade, the probiotic market has grown rapidly, both for foods and supplements intended to enhance wellness in healthy individuals. Different lactic acid bacteria (LAB), especially <i>Lactobacillus</i> spp., of different origins have already been used to develop commercial probiotic products. Nowadays, LAB new alternative sources, such as non-dairy fermented food products, are being exploited. One such source is Kombucha, a fermented low-alcohol beverage made of tea leaves. In this regard, we tested seven <i>Pediococcus</i> spp. strains isolated from a local industrial Kombucha for their biotechnological potential. Two, out of the seven isolates, identified as <i>Pediococcus pentosaceus</i> (L3) and <i>Pediococcus acidiliactici</i> (L5), were selected as successful candidates for the food industry, due to their probiotic and technological properties. In regard to their resistance in the gastro-intestinal tract, both selected strains were tolerant to a pH of 3.5, presence of 0.3% pepsin, and 0.5% bile salt concentration. On the antagonistic side, the fresh suspension of selected isolates had high inhibitory activity against pathogenic bacteria, such as <i>Salmonella enterica</i> Typhimurium, <i>Listeria monocytogenes, Listeria ivanovii, Bacillus cereus, Proteus hauseri,</i> and methicillin resistant <i>Staphylococcus aureus</i>. In addition, moderate to high inhibitory activity was noticed against foodborne molds (e.g., <i>Penicillium expansum</i> and <i>Penicillium digitatum</i>). These safety issues were supported by their negative hemolytic activity and good antioxidant potential (56–58%). Selected isolates were sensitive to ampicillin, penicillin, erythromycin, and lincomycin, while a broad range of other antibiotics were not effective inhibitors. On the technological side, both strains tolerated 5% NaCl and, during the freeze-drying process, had a good survival rate (86–92%). The selected <i>Pediococcus</i> strains have proven properties to be used for further development of functional products.
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