title: Purification and characterization of a phenoloxidase (laccase) from the lignin-degrading basidiomycete PM1 (CECT 2971).
abstract: A new lignin-degrading basidiomycete, strain PM1 (= CECT 2971), was isolated from the wastewater of a paper factory. The major ligninolytic activity detected in the basidiomycete PM1 culture supernatant was a phenoloxidase (laccase). This activity was produced constitutively in defined or complex media and appeared as two protein bands in native gel electrophoresis preparations. No enzyme induction was found after treatment with certain potential laccase inducers. Laccase I was purified to homogeneity by gel filtration chromatography, anion-exchange chromatography, and hydrophobicity chromatography. The enzyme is a monomeric glycoprotein containing 6.5% carbohydrate and having a molecular weight of 64,000. It has an isoelectric point of 3.6, it is stable in a pH range from 3 to 9, and its optimum pH is 4.5. The laccase optimal reaction temperature is 80 degrees C, the laccase is stable for 1 h at 60 degrees C, and its activity increases with temperature. Spectroscopic analysis revealed that the enzyme has four bound copper atoms, a type I copper, a type II copper, and a type III binuclear copper. The amino-terminal sequence of the protein is very similar to the amino-terminal sequences of laccases from Coriolus hirsutus and Phlebia radiata.
authors.affiliation: Instituto de Microbiología Bioquímica, Consejo Superior de Investigaciones Ceintíficas, Salamanca, Spain.
authors.forename: P M
authors.initials: PM
authors.lastname: Coll
authors.forename: J M
authors.initials: JM
authors.lastname: Fernández-Abalos
authors.forename: J R
authors.initials: JR
authors.lastname: Villanueva
authors.forename: R
authors.initials: R
authors.lastname: Santamaría
authors.forename: P
authors.initials: P
authors.lastname: Pérez
mesh_terms: D000595:Amino Acid Sequence; D001487:Basidiomycota / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D001673:Biodegradation, Environmental; D042845:Laccase; D008031:Lignin / Q000378:metabolism*; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D017386:Sequence Homology, Amino Acid; D014871:Water Microbiology
chemical_list: D008031:Lignin; D010088:Oxidoreductases; D042845:Laccase