AbstractThere is an increasing demand for green chemistry technologies that can cope with environmental waste management challenges. Agro-industrial residues are primarily composed of complex polysaccharides that support microbial growth for the production of industrially important enzymes such as ligninolytic enzymes. Schyzophyllum commune and Ganoderma lucidum were used alone, as well as mixed/co-culture, to produce crude ligninolytic enzymes extracts using corn stover and banana stalk as a substrate during solid state fermentation (SSF). In the initial screening, the extracted ligninolytic enzymes from S. commune produced using corn stover as the substrate showed higher activities of lignin peroxidase (1007.39 U/mL), manganese peroxidase (614.23 U/mL), and laccase (97.47 U/mL) as compared to G. lucidum and the mixed culture. To improve the production of ligninolytic enzymes by S. commune with solid state fermentation (SSF), physical factors such as pH, temperature, moisture, inoculum size, and incubation time were optimized by varying them simultaneously using response surface methodology (RSM) with a central composite design (CCD). The optimum SSF conditions were (for a 5 g corn stover substrate size): pH = 4.5; temperature = 35°C; inoculum size = 4 mL; and moisture content = 60%. Under optimum conditions, the activities of lignin peroxidase (LiP), manganese peroxidase (MnP), and laccase were 1270.40, 715.08, and 130.80 IU/mL, respectively.