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Related Experiment Videos

Purification and characterization of basic glucosyltransferase from Streptococcus mutans serotype c.

H Mukasa, A Shimamura, H Tsumori

    Biochimica Et Biophysica Acta
    |October 28, 1982
    PubMed
    Summary

    Streptococcus mutans secretes a glucosyltransferase enzyme that synthesizes 1,6-alpha-D-glucan from sucrose. This enzyme, composed of three isozymes, shows specific activity and optimal conditions for glucan synthesis.

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    Area of Science:

    • Microbiology
    • Biochemistry
    • Enzymology

    Background:

    • Streptococcus mutans is a key bacterium in dental caries development.
    • Glucosyltransferases (GTFs) produced by S. mutans synthesize glucans from sucrose, contributing to biofilm formation.
    • Understanding GTF structure and function is crucial for developing anti-caries strategies.

    Purpose of the Study:

    • To isolate and characterize the basic glucosyltransferase secreted by Streptococcus mutans Ingbritt (serotype c).
    • To elucidate the enzymatic properties and the structure of the synthesized glucan.

    Main Methods:

    • Enzyme purification using ethanol fractionation, DEAE Bio-Gel A chromatography, chromatofocusing, and preparative isoelectric focusing.
    • Enzyme characterization including molecular weight determination (SDS-PAGE), specific activity, optimal pH, and kinetic parameters (Km).

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  • Glucan structural analysis using gas-liquid chromatography-mass spectroscopy.

    • Main Results:

    • Three isozymes of basic glucosyltransferase (pI 8.1-8.4) were purified.
    • The enzyme has a molecular weight of 151,000, specific activity of 9.8 IU/mg, optimal pH of 6.5, and Km values of 7.1 µM for dextran and 4.3 mM for sucrose.
    • The synthesized glucan was identified as 1,6-alpha-D-glucan with 17.7% 1,3,6-branching.

    Conclusions:

    • Streptococcus mutans Ingbritt secretes a basic glucosyltransferase capable of de novo synthesis of branched glucans.
    • The characterized enzyme and its product provide insights into the mechanism of glucan formation by S. mutans.
    • Further research into this enzyme could lead to novel approaches for preventing dental caries.