In the present study, 33 strains of lactic acid bacteria (LAB) previously isolated from natural black Conservolea olives were assessed for their probiotic potential in vitro, as well as for their autoinducer-2 (AI-2) activity under standard growth conditions and the production of γ-aminobutyric acid (GABA). The probiotic tests included the in vitro resistance to low pH and resistance to bile salts, the evaluation of bile salt hydrolase activity, as well as safety tests regarding their possible haemolytic activity and their antimicrobial activity against pathogens. The results indicated that 17 strains were able to survive in low pH and in the presence of bile, with 15 of them also exhibiting partial bile salt hydrolase activity. None of the strains exhibited haemolytic activity or inhibited the growth of any of the examined pathogens. Moreover, the strains displayed generally low AI-2 activity under the growth conditions tested, regardless of the species. Interestingly, in contrast to what has been found in most foods, none of the isolates were found to produce GABA after 48 h of growth. The results from the AI-2 activity and extracellular GABA detection were considered as unexpected for LAB with probiotic attributes.