Cellobiose dehydrogenase from Schizophyllum commune: purification and study of some catalytic, inactivation, and cellulose-binding properties

Arch Biochem Biophys. 1998 May 1;353(1):37-46. doi: 10.1006/abbi.1998.0602.

Abstract

Cellobiose dehydrogenase (CDH) of Schizophyllum commune was purified to homogeneity. It is a glycoprotein with a molecular mass of 102, 000. Cellulosic substrates can serve as substrates for CDH. Cytochrome c, dichlorophenol-indophenol, ferricyanide, and oxygen can be reduced by the enzyme. CDH is stable in the pH range of 4-11 and up to 35 degrees C. The enzyme keeps active at high concentrations of H2O2. In the presence of cellobiose and Fe3+, incubation of CDH resulted in its inactivation and the degree of the inactivation was dependent mainly on the amount of CDH and cellobiose present. CDH has a distinct and specific affinity to cellulose and showed the strongest binding to acid-treated cellulose. The adsorption isotherm data fitted the Langmuir-type equation. The uv-visible spectra of the oxidized and reduced states of CDH showed a typical cytochrome b-type pattern. Addition of dithionite eliminated the adsorption between 440 and 500 nm, which indicates the presence of a flavin group in CDH.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carbohydrate Dehydrogenases / chemistry
  • Carbohydrate Dehydrogenases / isolation & purification*
  • Carbohydrate Dehydrogenases / metabolism*
  • Catalysis
  • Cations / pharmacology
  • Cellulose / metabolism
  • Chromatography, Gel
  • Chromatography, Ion Exchange
  • Electron Spin Resonance Spectroscopy
  • Enzyme Stability
  • Glycoproteins / chemistry
  • Hydrogen-Ion Concentration
  • Kinetics
  • Molecular Weight
  • Protein Binding
  • Schizophyllum / enzymology*
  • Substrate Specificity

Substances

  • Cations
  • Glycoproteins
  • Cellulose
  • Carbohydrate Dehydrogenases
  • cellobiose-quinone oxidoreductase