The present work was carried out to determine the best sources and concentrations of carbon and nitrogen in the production of laccases, total peroxidases and manganese peroxidase by Marasmiellus palmivorus VE111. Among the carbon and nitrogen sources used, the combination of glucose and casein led to higher production of laccases (5134 U/mL), total peroxidases (187 U/mL) and manganese peroxidase (57 U/mL). Central composite rotatable design (CCRD) was used to determine the optimum concentrations of glucose and casein to produce enzymes. The experiments with better results were scaled up in a 5 L bioreactor. It was verified that lower concentrations of glucose (5 and 10 g/L) stimulated higher activity of total peroxidases (1285 U/mL) and laccases (3420 U/mL), respectively, while a higher concentration of glucose (30 g/L) favoured the production of manganese peroxidase (59 U/mL). These data indicate high performance of M. palmivorus VE111 in the production of ligninolytic enzymes.