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AbstractThis study examined the effect of lactulose, galactooligosaccharide, fructooligosaccharide, inulin, and β-glucan on the probiotic strain Lactobacillus delbrueckiisubsp. bulgaricusL14, cultivated in an in vitrogastrointestinal system model. We analyzed the degree of hydrolysis of the studied prebiotic oligosaccharides in condition of simulated gastric fluid. The results showed that lactulose had the highest resistance, galactooligosaccharide underwent hydrolysis, and fructooligosaccharide was the most sensitive. Among the polysaccharides, fructose was released from inulin and glucose from β-glucan. Short-chain oligosaccharides and metabolites derived from studied prebiotic oligosaccharides and polysaccharides, supported the growth of probiotic strain L14, which showed the highest growth with fructooligosaccharides and β-glucan as carbohydrate sources. The profile of the activated enzymes secreted by the probiotic strain L14, indicated their inducible character. Beta-galactosidase was activated in the presence of lactulose and GalOS, inulinase was activated in the presence of inulin and fructooligosaccharides, and β-glucosidase was activated in the presence of β-glucan fragments. Analysis of the produced organic and short-chain fatty acids showed that the typical representative of the homofermentative lactobacilli Lb. delbrueckiisubsp. bulgaricusL14 changes its metabolism from a homofermentative to a heterofermentative type, best expressed in the presence of lactulose, galactooligosaccharide, and β-glucan.