Lactobacillus rhamnosus E/N is a probiotic bacterium, which synthesizes exopolysaccharides (EPS) with significant bifidogenic and antioxidant activities. The sugar composition of the EPSs produced depended on carbohydrates used as a carbon source in the growth media. Five Bifidobacterium strains were tested in vitro for their ability to utilize all the EPSs studied. The highest bifidogenic activity was revealed by EPSs obtained from Lactobacillus cultures supplemented with Gal, Lac, and Mal as the only carbon source, while significant antioxidant effects were observed in EPSs isolated from growth media enriched with galactose, lactose, and sucrose.