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Related Experiment Videos

Microsomal chitinase activity from Candida albicans.

K Dickinson1, V Keer, C A Hitchcock

  • 1Department of Microbiology, University of Leeds, U.K.

Biochimica Et Biophysica Acta
|January 23, 1991
PubMed
Summary
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Microsomal chitinase in Candida albicans yeast cells exhibits zymogenic properties and shares characteristics with its cytosolic counterpart. This enzyme

Area of Science:

  • Biochemistry
  • Mycology
  • Enzymology

Background:

  • Chitinase (E.C. 3.2.1.14) plays a role in fungal cell wall biology.
  • Understanding chitinase localization and properties is crucial for targeting fungal pathogens.
  • Candida albicans is a significant opportunistic fungal pathogen.

Purpose of the Study:

  • To characterize microsomal chitinase activity in Candida albicans.
  • To investigate the properties and localization of microsomal chitinase.
  • To explore the potential of microsomal chitinase as an antifungal target.

Main Methods:

  • Enzyme activity assays on microsomal fractions from Candida albicans.
  • Washing protocols to assess enzyme activation and stability.
  • Pre-incubation with trypsin to evaluate zymogenic properties.

Related Experiment Videos

  • Determination of kinetic parameters (Km) and inhibition constants (Ki).
  • Solubilization studies using detergents.
  • Density gradient centrifugation for enzyme localization.

    • Main Results:

    • Microsomal chitinase activity was low in unwashed fractions but increased with washing and trypsin activation.
    • Microsomal chitinase exhibited kinetic and inhibition properties similar to cytosolic chitinase.
    • Detergents differentially solubilized and activated the enzyme.
    • Chitinase was distributed broadly within the microsomes, not exclusively bound to one component.

    Conclusions:

    • Microsomal chitinase in Candida albicans is likely zymogenic and shares properties with cytosolic chitinase.
    • The enzyme's distribution suggests it is not restricted to a single membrane component.
    • Microsomal chitinase represents a potential target for developing specific antifungal agents.