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Related Experiment Videos

Dynamic reactivities of dextransucrase.

S L Ditson, S M Sung, R M Mayer

    Archives of Biochemistry and Biophysics
    |August 15, 1986
    PubMed
    Summary

    Dextransucrase exhibits novel reactivity, catalyzing oligosaccharide depolymerization and glucosyl residue transfer to various acceptors, including water and fructose, forming glucose and sucrose, respectively.

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    Area of Science:

    • Enzymology
    • Carbohydrate Chemistry
    • Biochemistry

    Background:

    • Dextransucrase from Streptococcus sanguis ATCC 10558 is an enzyme that synthesizes dextran from sucrose.
    • Previous studies utilized pulse-chase experiments to investigate enzyme-bound oligosaccharides.

    Purpose of the Study:

    • To investigate the unexpected reactivity of immobilized dextransucrase previously charged with labeled sucrose.
    • To elucidate the mechanism of glucosyl residue transfer and oligosaccharide metabolism by dextransucrase.

    Main Methods:

    • Immobilization of dextransucrase on hydroxylapatite.
    • Pulsing the immobilized enzyme with labeled sucrose.
    • Pulse-chase experiments using low concentrations of unlabeled sucrose.
    • Analysis of radioactive labeling at the reducing termini of enzyme-bound oligosaccharides.
    • Enzymatic assays with various acceptors like maltose, water, and fructose.

    Main Results:

    • Contrary to previous findings, low concentrations of unlabeled sucrose in the chase led to an increase in radioactivity at the reducing termini.
    • Dextransucrase was shown to depolymerize bound oligosaccharides.
    • Individual glucosyl residues were transferred to acceptors (maltose, water, fructose) forming trisaccharides, glucose, and sucrose.
    • Oligosaccharides were slowly released from the enzyme.
    • The reverse reaction, sucrose synthesis from transferred glucose residues and fructose, was also observed.

    Conclusions:

    • Dextransucrase exhibits complex reactivity beyond dextran synthesis, including oligosaccharide depolymerization and glucosyl transfer.
    • The enzyme can act as a glucosyltransferase, transferring residues to water (hydrolysis) or other carbohydrates.
    • The enzyme also demonstrates the ability to synthesize sucrose from transferred glucose residues and fructose.

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