In this work we review the extraordinary biotechnological potential of two glycosyltransferases, cyclodextrin glucanotransferase and dextransucrase, especially their utility in the synthesis of oligosaccharides. Both enzymes are non-Leloir transferases that require neither co-factors nor activated substrates, as they directly employ the free energy of cleavage of starch and sucrose, respectively. Cyclodextrin glucanotransferase is able to produce cyclodextrins from starch. In the presence of appropriate acceptors (e.g., carbohydrates), this enzyme furnishes oligosaccharides containing α(1[Formula: see text]4) bonds. Thus, we have found that glucose, maltose, and sucrose readily serve as acceptors to form the corresponding [Glc-α(1[Formula: see text]4)]n- oligosaccharides, with the degree of polymerization being controlled by the starch:acceptor ratio. The ability of other sugars and related compounds to act as acceptors is also reviewed. Dextransucrase is a glycansucrase that synthesizes dextran using sucrose as glucosyl donor. The formation of dextrans can be quantitatively replaced with the formation of novel oligosaccharides by adding alternative carbohydrate acceptors to the reaction medium. With the dextransucrase from Leuconostoc mesenteroides B-1299, we have investigated the synthesis of gluco- oligosaccharides containing α(1[Formula: see text]2) bonds using methyl 1-O-α-D-glucopyranoside as the acceptor. These products constitute a class of nondigestible nutraceutical oligosaccharides with prebiotic properties relating to the stabilization and enhancement of gastrointestinal tract flora, and are being increasingly used by the food industry.Key words: glycansucrases, cyclodextrin glucanotransferase, cyclodextrin glucosyltransferase, dextransucrase, acceptor products, gluco-oligosaccharides, malto-oligosaccharides, coupling sugar, nutraceuticals, functional foods, prebiotics.