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Related Experiment Videos

Genes from Cellvibrio mixtus encoding beta-1,3 endoglucanase.

H Sakellaris1, J M Pemberton, J M Manners

  • 1Department of Microbiology, University of Queensland, St. Lucia, Australia.

Applied and Environmental Microbiology
|October 1, 1990
PubMed
Summary

Researchers cloned two beta-1,3 glucanase genes from Cellvibrio mixtus, yielding distinct enzymes. One enzyme degraded yeast cell walls and various beta-1,3 glucans, while the other targeted laminarin specifically.

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Area of Science:

  • Microbiology
  • Enzymology
  • Molecular Biology

Background:

  • Beta-1,3 glucanases are enzymes that break down beta-1,3 glucans, important components of fungal cell walls and storage polysaccharides.
  • Cellvibrio mixtus is a soil bacterium known to produce extracellular enzymes, including glucanases.

Purpose of the Study:

  • To clone and characterize genes encoding beta-1,3 glucanase activity from Cellvibrio mixtus.
  • To investigate the substrate specificities and enzymatic properties of the cloned beta-1,3 glucanases.

Main Methods:

  • Gene cloning using bacterial transformation and plasmid isolation.
  • Endonuclease restriction mapping to differentiate clones.
  • Enzyme activity assays with various beta-1,3 glucan substrates (laminarin, zymosan, pachyman, yeast cell walls).

Related Experiment Videos

  • Analysis of degradation products (laminarobiose, laminarotriose, laminarotetraose).
  • Radiolabeling and identification of translated gene products.

    • Main Results:

    • Two distinct genes, cwd and lam, encoding beta-1,3 glucanase activity were successfully cloned.
    • The cwd gene product degraded yeast cell walls, laminarin, zymosan, and pachyman.
    • The lam gene product specifically degraded laminarin.
    • Both enzymes exhibited endohydrolytic activity on laminarin, producing laminarobiose, laminarotriose, and laminarotetraose.

    Conclusions:

    • Cellvibrio mixtus possesses at least two distinct beta-1,3 glucanase genes with differing substrate specificities.
    • The cloned enzymes, CWD and LAM, offer potential applications in degrading fungal biomass or modifying glucan-containing materials.