Cellobiose dehydrogenases of Sporotrichum (Chrysosporium) thermophile

Eur J Biochem. 1991 May 23;198(1):43-52. doi: 10.1111/j.1432-1033.1991.tb15984.x.

Abstract

Both cellobiose dehydrogenases of Sporotrichum (Chrysosporium) thermophile, ATCC 42464, obtained after fractionation with DEAE-Trisacryl chromatography and named cellobiose dehydrogenase I and II have been purified to homogeneity by different chromatographic techniques. Both enzymes are slightly glycosylated flavocytochrome-b proteins with similar catalytic properties but with distinct molecular masses (91 kDa and 192 kDa for enzymes I and II, respectively) and isoelectric point (4.1 versus 3.45). Examination by SDS/PAGE clearly showed that the larger enzyme II is a homodimer, whose subunit is close to, but different from dehydrogenase I which is homogeneous by this technique. After limited digestion of both enzymes with papain, two main fractions with residual activity are formed, one carrying the heme, the other being the flavin component; each fraction is characterized by its particular chromatographic behaviour. The flavin carrying component shows an atypical (for flavoprotein) three-banded spectrum indicative of the presence of a flavin derivative. Both enzymes react very slowly with oxygen clearly forming some superoxide radicals and possibly hydrogen peroxide. Cellobiose and other cellodextrins are oxidized at their reducing glycosyl moiety to the corresponding aldonic acid. With the use of the autooxidable phenazinemethosulphate, cellulose (either in a hydrated form or crystalline) is also oxidized at free reducing ends so that appreciable amounts of cellobionic acid are released upon enzymatic hydrolysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / analysis
  • Carbohydrate Dehydrogenases / chemistry*
  • Carbohydrate Dehydrogenases / isolation & purification
  • Chromatography, High Pressure Liquid
  • Electrophoresis, Polyacrylamide Gel
  • Flavins / analysis
  • Hydrogen Peroxide / analysis
  • Isoelectric Focusing
  • Molecular Weight
  • Oxidation-Reduction
  • Spectrum Analysis
  • Sporothrix / enzymology*
  • Substrate Specificity
  • Superoxides / analysis

Substances

  • Amino Acids
  • Flavins
  • Superoxides
  • Hydrogen Peroxide
  • Carbohydrate Dehydrogenases
  • cellobiose-quinone oxidoreductase