Abstract

• In this research, laccase isolated from fungal source of mushroom, Trametes versicolor, white rot fungi in solid state fermentation. Laccase was purified by ammonium sulphate fractionation (20 % and 70 %) and dialysis followed by Sephadex G-100 gel filtration chromatography In each purification step protein content was determined by Biuret method using Bovine Serum Albumin as standard at 560 nm and laccase activity was determined by gauaicol assay method at 450 nm The extracellular laccase from T. vesicolor was purified to 12.72 fold. The purity of laccase was confirmed by SDS-PAGE as single band. The molecular weight of purified laccase was found to be 60.26 kDa. The immobilization of laccase was carried out by gel entrapment technique using sodium alginate- gelatin- agar mixed gel. The highest laccase activities were found at pH 5 for free laccase at 40 oC and pH 6 for the immobilized laccase. The optimal temperatures of free laccase and the immobilized laccase were 40 oC and 45 oC, respectively, showing the improvement in thermal stability of immobilized laccase. The reaction order of both free and immobilized laccase catalyzed reactions was first order reaction.