Dynamic reactivities of dextransucrase.
Dextransucrase, from Streptococcus sanguis ATCC 10558, was immobilized on hydroxylapatite and was "charged" in short pulses with labeled sucrose, as previously described [V. K. Parnaik, G. A. Luzio, D. A. Grahame, S. L. Ditson, and R. M. Mayer (1983) Carbohydr. Res. 121, 257-268]. The "charged" enzyme has been shown to contain both bound glucose and gluco-oligosaccharides. The reactivity of this form of the enzyme has been studied, and shown to have unexpected behavior. Earlier pulse-chase experiments [J. F. Robyt, B. K. Kimble, and T. F. Walseth (1979) Arch. Biochem. Biophys. 165, 634-640; S. L. Ditson and R. M. Mayer (1984) Carbohydr. Res. 126, 170-175], carried out with high concentrations of unlabeled sucrose in the chase, resulted in a rapid decrease in isotope at the reducing termini of enzyme-bound oligosaccharides. However, in the present work, in which the pulsed enzyme was chased with low concentrations of unlabeled sucrose, we observed an increase in the radioactive reducing termini. The possibility that this was due to the enzymatic hydrolysis of dextran has been ruled out. Data presented demonstrate that the enzyme catalyzes the depolymerization of the bound oligosaccharides. Individual glucosyl residues of the oligosaccharides are transferred to acceptors, such as added maltose to form a trisaccharide, or water to form glucose. Similarly, the glucosyl residues can be transferred to added fructose to form sucrose. The studies also provide evidence that the oligosaccharides are slowly released from the enzyme. The ability of the enzyme to catalyze the reverse of the glucosyl transfer reaction involving acceptors was also examined. It was observed that glucose residues transferred by dextransucrase to an acceptor can also be removed to produce sucrose when fructose is added.
D004800:Enzymes, Immobilized; D005632:Fructose / Q000378:metabolism; D005964:Glucosyltransferases / Q000378:metabolism*; D006602:Hexosyltransferases / Q000378:metabolism; D011108:Polymers; D013298:Streptococcus sanguis / Q000201:enzymology*; D013395:Sucrose / Q000378:metabolism
D004800:Enzymes, Immobilized; D011108:Polymers; D005632:Fructose; D013395:Sucrose; D005964:Glucosyltransferases; D006602:Hexosyltransferases; C020025:dextransucrase