Purification and characterization of laccases from the white-rot basidiomycete Dichomitus squalens

Frédéric H. Périé, G. Vijay Bhasker Reddy, Ninian Blackburn, Michael H. Gold

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Two chromatographic forms of laccase c1 and c2 were purified approximately 225-fold from the extracellular culture fluid of ligninolytic cultures of Dichomitus squalens, using DEAE-Sepharose and Mono-Q fast protein liquid chromatography. Each homogeneous laccase (c1 and c2) has a molecular mass of approximately 66 kDa as determined by SDS-PAGE. Both forms are glycoproteins, and each contains four copper atoms per molecule of protein. The first 20 amino acids of the N-terminal sequences of these two laccases are identical and are similar to those of laccases from other lignin- degrading fungi. The electronic absorption spectra of these laccases exhibit bands at 610 and 330 nm, indicative of type I and type III copper. The EPR spectrum of laccase c1 exhibits bands indicative of type I and type II copper. Each laccase oxidizes a variety of phenolic substrates, has a pH optimum of 3.0 for the oxidation of 2,6-dimethoxyphenol, and is inhibited strongly by fluoride and azide.

Original languageEnglish (US)
Pages (from-to)349-355
Number of pages7
JournalArchives of Biochemistry and Biophysics
Volume353
Issue number2
DOIs
StatePublished - May 15 1998

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Laccase
Basidiomycota
Purification
Copper
Azides
Lignin
Extracellular Fluid
Liquid chromatography
Molecular mass
Fungi
Fluorides
Liquid Chromatography
Sepharose
Paramagnetic resonance
Polyacrylamide Gel Electrophoresis
Absorption spectra
Glycoproteins
Proteins
Amino Acids
Atoms

Keywords

  • Basidiomycete
  • Dichomitus squalens
  • Fungi
  • Lignin degradation
  • Multicopper oxidase
  • Phanerochaete chrysosporium

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Purification and characterization of laccases from the white-rot basidiomycete Dichomitus squalens. / Périé, Frédéric H.; Reddy, G. Vijay Bhasker; Blackburn, Ninian; Gold, Michael H.

In: Archives of Biochemistry and Biophysics, Vol. 353, No. 2, 15.05.1998, p. 349-355.

Research output: Contribution to journalArticle

Périé, Frédéric H. ; Reddy, G. Vijay Bhasker ; Blackburn, Ninian ; Gold, Michael H. / Purification and characterization of laccases from the white-rot basidiomycete Dichomitus squalens. In: Archives of Biochemistry and Biophysics. 1998 ; Vol. 353, No. 2. pp. 349-355.
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