Purification, partial characterization, and reactivity with aromatic compounds of two laccases from Marasmius quercophilus strain 17.
Two isozymes of laccase were obtained from an induced liquid culture of Marasmius quercophilus with p-hydroxybenzoic acid as the inducer. Both the constitutive and the induced isozyme have a molecular mass of 60 kDa as determined by polyacrylamide gel electrophoresis. Using isoelectric focusing, we found three isozymes with the constitutive enzyme (pI 4, 4.2, 4.4) and four of the induced form (pI 4.75, 4.85, 4.95, 5.1). We observed certain differences between these two isozymes; the specific activity of the induced isozyme was twice as high, and two optimum pH levels (5 and 6) were observed with the induced isozyme (only one, pH 5, for the constitutive isozyme). However, both of these enzymes have the same thermal stability and the same temperature for their highest activity (80 degrees C). Furthermore, the reactivity of both these enzymes with aromatic compounds was similar. The use of mediators extended the oxidized substrate range of the laccases studied. Various products of degradation were observed, depending on the mediator used. When laccase was used alone, the decrease of the signal corresponding to the aromatic cycle, without any formations of other peaks at different wavelengths, suggested polymerisation of aromatic compounds.
Farnet
A M
AM
Laboratoire de Microbiologie, Service 452, Institute Méditerranéen d'Ecologie et de Paléoécologie, Faculté des Sciences et Techniques de St. Jérôme, Marseille, France.
Criquet
S
S
Tagger
S
S
Gil
G
G
Le Petit
J
J
D000363:Agaricales / Q000201:enzymology* / Q000254:growth & development; D002851:Chromatography, High Pressure Liquid; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000302:isolation & purification; D042845:Laccase; D010088:Oxidoreductases / Q000302:isolation & purification* / Q000378:metabolism*; D010636:Phenols / Q000378:metabolism*
D003470:Culture Media; D007527:Isoenzymes; D010636:Phenols; D010088:Oxidoreductases; D042845:Laccase