Assimilation of cholesterol and prebiotic substrates are functionally properties of prebiotics strains that can further contribute to their desirable effects on the health of the host. Selected strains of Lactococcus and Enterococcus species satisfied the general criteria for the selection of probiotics strains. In order to exclude the possibility of spread of antibiotic resistance, we examined the sensitivity of lactic acid bacteria (LAB) to 5 antibiotics. The investigation shown that the strain Enterococcus faecium ZG1-54 was sensitive to all six tested antibiotics, while strains Lactococcus lactis BGAL1-1 and Lactococcus lactis subsp. lactis BGLE1-6 showed resistance to erythromycin and a strain Lactococcus lactis ZG5-9 to tetracycline and vancomycin. In order to achieve stronger synbiotic effect, we have studied the assimilation of prebiotic substrates (mannitol, lactulose, inulin and raftiloze) with selected probiotic strains BGAL1-1, BGLE1-6, ZG1-54 and ZG5-9. Growth of selected strains, the changes of pH and synthesis of lactic acid were the highest during cultivation in nutrient media with mannitol and lactulose as a prebiotic substrates. For the determination of basic and acidic properties of microbial cells has been used method of microbial adhesion in 3 solvents. A large percentage adhesion of cell BGAL1-1 to hexane indicate to their hydrophobicity and the results of microbial adhesion to chloroform show an affinity of the investigated BMK to acidic solvent. It refers to their electron-donor properties. Selected probiotic strains did not show antagonism towards each other and can grow as paired in the same culture medium. Selected strains have the ability to assimilate cholosterol, which could be another desirable characteristic of probiotic bacteria.