Purification and Properties of Laccase Produced by Coriolus hirsutus in Solid State Fermentation

Abstract:

In this paper,purification and properties of laccase produced by Coriolus hirsutus in solid state fermentation were studied.Crude extract of laccase from Coriolus hirsutus was collected after solid state fermentation.The laccase was purified to electrophoretic homogeneity by the serials of steps of centrifugation,filtration,ammonium sulfate precipitation,ion-exchange chromatography on DEAE-Sepharose fast flow and gel filtration on sephadex G-100.Purification of 15.53 fold was achieved with an overall yield of 51.21%.Its molecular weight was estimated to be about 62 600 by SDS-PAGE.Studies on the properties of the laccase showed that the optimum temperature was 50 ℃ and the optimum pH were 4.0 in citrate buffer and 3.0 in sodium acetate buffer.The laccase catalyzed the oxidation of ABTS and catechol with a michaelis constant of 0.093 mmol/L and 3.71 mmol/L,respectively.