Purification And Characterization of Trametes hirsute DSMZ No.5072 Laccase

Abstract

In this study, the Production of laccase enzyme from Trametes hirsuta strain (DSMZ No.5072) in the 2L fermenter using different culturing media had been achieved. The production of laccase was also induced by using different concentrations of Copper sulphate, 2, 5 Xylidine and Gallic acid as inducers. The maximum laccase activitiy observed during T. hirsuta growth in the presence of various inducers was 5.89 U ml -1. Laccase purification was performed by precipitated the enzymes with ammoniam sulphate, saturation 90%, followed by gel filtration chromatography using Sephadex-G25 and more purified by ion-exchanger (DEAE-Sephadex). The obtained enzyme was concentrated by ultra filtration membrane and analyzed by SDS-PAGE. The activity of home prepared laccase and the commercial laccase from Trametes versicolor were carried out in different media (aqueous, reverse micelles, co-solvent, ionic liquid and ternary systems) depend on oxidation of ABTS as an enzyme substrate. The optimum pH for the laccase activity of the two fungal laccases was observed at acidic pH values close to (pH 3.5-4.6) while the optimum temperature was 70ºC.