EXTRACTION AND PURIFICATION OF EXTRACELLULAR LACCASE FROM WILD, MUTANTS AND HYBRID STRAINS OF TWO WHITE-ROT FUNGUS AND ITS APPLICATIONS IN DECOLOURIZATION AND LIGNINOLYSIS

摘要

Extracellular laccases were extracted from a 5-day old submerge cultures of the wild, mutants and hybrid of Lentinus subnudus. Mutants were generated by exposure of the wild strain of L. subnudus to ultraviolet radiation (? = 280 nm) at specific time intervals while the hybrid was produced by cross-breeding L. subnudus with L. edodes. The crude enzyme was fractionated with 80% ammonium sulphate and further purified on DEAE column. The laccase has a molecular weight of about 45 KDa. Purification yield on DEAE column gave the highest purification yield of 23.25% in SWT and least in SHT (5.29%). Its potentials in decolourization of 2, 6-dichlorophenol-indophenol dye at different pH conditions were investigated. Five out of the six fungal strains tested gave significant (P<0.05) percentage decolourization (≥43.94%) at pH 8. The fungus was further studied for their ability in degrading wheat and paddy straws. The solid substrate fermentation was inoculated with two pieces (0.6cm diameter) mycelial agar blocks of each of the fungal strains, supplemented with 30mg/100g sucrose, 24mg/100g KNO3 and 60mg/100g CaCO3. The periodic reduction in weight of the solid substrate medium and enzymatic activity of laccase for each of the fungal strains was assessed. Therefore, the ability of the wild, mutants and hybrid of L subnudus strains to produce laccase enzyme shows their significant potential in textile industry, especially in decolourization of dye and bioconversion of lignocellulosic wastes.