Hydrolase and transferase activities of the beta-1,3-exoglucanase of Candida albicans

Eur J Biochem. 1999 Aug;263(3):889-95. doi: 10.1046/j.1432-1327.1999.00581.x.

Abstract

The exo-beta-1,3-glucanase of Candida albicans (Exg) has a marked specificity for beta-1,3-glucosidic linkages as judged by the kinetic constants for p-nitophenyl beta-glucoside, beta-linked disaccharides of glucose (laminaribiose, gentiobiose, and cellobiose), oligosaccharides of the laminari series, laminarin and pustulan. The kcat/Km ratios for a series of laminari oligosaccharides from -biose to -heptaose showed that Exg has an extended substrate-binding site which contains at least five binding sites for sugar residues. Binding at position +2 (the third sugar residue) increases the kcat twofold while positions +3 and +4 lower the Km value further and thereby increase the catalytic efficiency. Exg catalyses an efficient transglucosylation reaction with high concentrations of laminari-oligosaccharides which specifically form beta-1,3 linkages and with yields up to 50%. The rate of the transglucosylation is concentration-dependent and can be more than 10 times faster than the hydrolytic reaction with excess donor substrates such as laminaritriose and laminarihexaose. The kinetics of Exg and the predicted substrate-binding site for up to five sugar residues are consistent with a recent structural analysis of the enzyme-binding site.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • Candida albicans / enzymology*
  • Carbohydrate Sequence
  • Glucan 1,3-beta-Glucosidase*
  • Glycoside Hydrolases / chemistry
  • Glycoside Hydrolases / metabolism*
  • Hydrolases / metabolism*
  • Kinetics
  • Molecular Sequence Data
  • Oligosaccharides / chemistry
  • Oligosaccharides / isolation & purification
  • Oligosaccharides / metabolism*
  • Substrate Specificity
  • Transferases / metabolism*

Substances

  • Oligosaccharides
  • Transferases
  • Hydrolases
  • Glycoside Hydrolases
  • Glucan 1,3-beta-Glucosidase
  • beta-1,3-exoglucanase