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There are differences in the extracellular enzymes produced from species of wood-rotting fungi and their activity due to variation in nutritional conditions such as carbon sources and nitrogen sources, as well as environmental conditions such as incubation temperature and pH. In this study, to determine the methods to promote the secretion of especially cellulase with high activity among the extracellular enzymes of wood-damaging fungi, the optimal nutrient sources and incubation conditions for the production of cellulase with high activity were investigated using response surface methodology based on a broth culture from Acanthophysium sp. KMF001, a novel strain of wood-damaging fungus. The nutrient sources that were optimal for the cellulase production with high activity from Acanthophysium sp. KMF001 were cellulose as a carbon source and tryptone:yeast extract (7:3) as a nitrogen source. The optimal incubation conditions were a temperature of 30 °C and a pH of 6. The optimal concentrations of carbon and nitrogen sources were cellulose at 31.1 g∙L-1 and tryptone:yeast extract (7:3) at 15 g∙L-1, with pH of 5.9.