Production of hydroxyl radical by the synergistic action of fungal laccase and aryl alcohol oxidase.
A mechanism for the production of hydroxyl radical (*OH) during the oxidation of hydroquinones by laccase, the ligninolytic enzyme most widely distributed among white-rot fungi, has been demonstrated. Production of Fenton reagent (H2O2 and ferrous ion), leading to *OH formation, was found in reaction mixtures containing Pleurotus eryngii laccase, lignin-derived hydroquinones, and chelated ferric ion. The semiquinones produced by laccase reduced both ferric to ferrous ion and oxygen to superoxide anion radical (O2*-). Dismutation of the latter provided the H2O2 for *OH generation. Although O2*- could also contribute to ferric ion reduction, semiquinone radicals were the main agents accomplishing the reaction. Due to the low extent of semiquinone autoxidation, H2O2 was the limiting reagent in Fenton reaction. The addition of aryl alcohol oxidase and 4-methoxybenzyl alcohol (the natural H2O2-producing system of P. eryngii) to the laccase reaction greatly increased *OH generation, demonstrating the synergistic action of both enzymes in the process.
Guillén
F
F
Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain. guillen@cib.csic.es
Gómez-Toribio
V
V
Martínez
M J
MJ
Martínez
A T
AT
D000429:Alcohol Oxidoreductases / Q000378:metabolism*; D005656:Fungal Proteins / Q000378:metabolism; D017665:Hydroxyl Radical / Q000378:metabolism*; D042845:Laccase; D010088:Oxidoreductases / Q000378:metabolism*; D020076:Pleurotus / Q000378:metabolism*
D005656:Fungal Proteins; D017665:Hydroxyl Radical; D010088:Oxidoreductases; D000429:Alcohol Oxidoreductases; C077759:aryl-alcohol oxidase; D042845:Laccase