The objective of the investigation conducted was to determine the growth and acidifying activity of Lactobacillus species in a medium with fructooligosaccharides and galactosylpolyols preparations. The bacteria cultures were developed using an MRS nutrient with glucose as a source of carbon. The glucose was substituted by fructooligosaccharides and galactosylpolyols in the MRS medium. An HPLC method was used to determine the content of lactic and acetic acids as produced by Lactobacillus species. The experiments proved that the bacterial strains of Lactobacillus species grew on a medium with added preparations of fructooligosacchaides and galactosylopolyols. The biomass yield of Lactobacillus sp. in a medium with fruktooligosaccharides varied from 0.57 g/l to 1.21 g/l, and as for the medium with glactosylopolyols: from 0.37 g/l to 0.63 g/l. Depending on the species types and the source of carbon, the pH values ranged between 4.7 and 5.5 against the initial value of 5.7. The molar concentration ratio between the acetic and the lactic acids produced by Lactobacilus species ranged from 0.02 to 0.3 and had a tendency to increase along with the rise in the degree of polymerization of oligomers. The galactosylpolyols are a source of carbon for the bacterial strains of Lactobacillus and their acidifying activity is weaker than the same activity of preparations of fructooligosaccharides.