Biochemical and molecular characterization of a laccase from Marasmius quercophilus.
The basidiomycete Marasmius quercophilus is commonly found during autumn on the decaying litter of the evergreen oak (Quercus ilex L.), a plant characteristic of Mediterranean forest. This white-rot fungus colonizes the leaf surface with rhizomorphs, causing a total bleaching of the leaf. In synthetic liquid media, this white-rot fungus has strong laccase activity. From a three-step chromatographic procedure, we purified a major isoform to homogeneity. The gene encodes a monomeric glycoprotein of approximately 63 kDa, with a 3.6 isoelectric point, that contains 12% carbohydrate. Spectroscopic analysis of the purified enzyme (UV/visible and electron paramagnetic resonance, atomic absorption) confirmed that it belongs to the "blue copper oxidase" family. With syringaldazine as the substrate, the enzyme's pH optimum was 4.5, the optimal temperature was 75 degrees C, and the K(m) was 7.1 microM. The structural gene, lac1, was cloned and sequenced. This gene encodes a 517-amino-acid protein 99% identical to a laccase produced by PM1, an unidentified basidiomycete previously isolated from wastewater from a paper factory in Spain. This similarity may be explained by the ecological distribution of the evergreen oak in Mediterranean forest.
Dedeyan
B
B
Laboratoire de Bioinorganique Structurale, CNRS UMR 6517, Faculté des Sciences de St Jérôme, 13397 Marseille, Cedex 20, France.
Klonowska
A
A
Tagger
S
S
Tron
T
T
Iacazio
G
G
Gil
G
G
Le Petit
J
J
D000363:Agaricales / Q000201:enzymology / Q000235:genetics*; D000595:Amino Acid Sequence; D003488:Cyanogen Bromide; D005800:Genes, Fungal; D006023:Glycoproteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D007527:Isoenzymes / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D042845:Laccase; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D020539:Sequence Analysis, Protein; D013379:Substrate Specificity
D006023:Glycoproteins; D007527:Isoenzymes; D010088:Oxidoreductases; D042845:Laccase; D003488:Cyanogen Bromide