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An intermediate in cyclic beta 1-2 glucan biosynthesis.

A Zorreguieta, R A Ugalde, L F Leloir

    Biochemical and Biophysical Research Communications
    |January 16, 1985
    PubMed
    Summary
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    Agrobacterium tumefaciens synthesizes cyclic beta 1-2 glucan attached to proteins. This process involves UDP-glucose and results in the release of free glucan, a key bacterial virulence factor.

    Area of Science:

    • Microbiology
    • Biochemistry
    • Molecular Biology

    Background:

    • Agrobacterium tumefaciens is a plant pathogen known for its ability to transfer DNA.
    • Cyclic beta 1-2 glucan is a crucial exopolysaccharide involved in bacterial virulence and biofilm formation.
    • The biosynthesis pathway of cyclic beta 1-2 glucan in A. tumefaciens has been partially elucidated.

    Purpose of the Study:

    • To investigate the mechanism of cyclic beta 1-2 glucan synthesis in Agrobacterium tumefaciens.
    • To identify the role of UDP-glucose in the formation of cyclic beta 1-2 glucan and associated compounds.
    • To elucidate the relationship between insoluble precursors and the final cyclic glucan product.

    Main Methods:

    • Incubation of UDP-[14C]Glc with A. tumefaciens inner membranes.

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  • Analysis of reaction products using proteolysis, acid hydrolysis, and alkaline treatment (beta-elimination).
  • Competition assays with non-radioactive UDP-Glc to assess pathway regulation.

    • Main Results:

    • Radioactive cyclic beta 1-2 glucan and trichloroacetic acid-insoluble compounds were formed from UDP-[14C]Glc.
    • Partial acid hydrolysis of both products yielded identical saccharide fragments.
    • Excess non-radioactive UDP-Glc inhibited glucan formation and reduced radioactivity in the precipitate.
    • Alkaline treatment released approximately 30% of free saccharides from the insoluble compounds.

    Conclusions:

    • Cyclic beta 1-2 glucan chains are synthesized while attached to a proteinaceous component.
    • The insoluble precursor represents a protein-glycan complex.
    • The final step involves the release of free cyclic beta 1-2 glucan from the protein.