Optimized production, purification and molecular characterization of fungal laccase through Alternaria alternata

Abstract

Abstract Objective Industrial effluents and agriculture biomass are main environmental hazards which are facing by developing country like Pakistan. Along with various other industrial applications, laccases are also involved in the oxidation of various industrial hazardous compounds to detoxify them. This study was designed to produce and purify laccase from ascomyceteous fungi, i.e. Alternaria alternata through solid stat fermentation. Materials and methods Abundantly available Sarkanda grass “Saccharum spontaneum” was used as agro-waste substrate for laccase production from fungus A. alternata. Previously only white rot fungi are familiar for laccase production and almost no work has been done on laccase production by A. alternata. In this research work, different physical and chemical parameters were optimized for maximum laccase production through solid state fermentation (SSF). Results Enzyme was purified and its molecular weight was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Maximum laccase activity (21.87±0.0115 unit/mL) was detected on 7th day of incubation having pH 5 of the medium at 35°C. None of the added metal ions increased laccase production. Galactose and “yeast extract” used as optimum carbon and nitrogen source for highest laccase production. Conclusion A monomeric protein (laccase) having approximately 51 kDa molecular weight obtained after SDS-PAGE.