This study was conducted on crude enzyme from the novel strain Acanthophysium sp. KMF001 using a mediator (surfactant). The surfactant was applied to the steam-exploded pretreated domestic wood biomass, and response surface methodology (RSM) was conducted to determine the optimum conditions for saccharification using the optimum substrate concentration, enzyme concentration, and surfactant concentration. Steam-explosion of Korean oak (25 kgf/cm2) for 7 min showed a maximum-predicted saccharification of approximately 99.9% at 7.0% substrate concentration, 37.5 FPU (filter paper units) enzyme concentration, and 475.8 mg/g-glucan surfactant (polysorbate 80) concentration. Steam-explosion of red pine (25 kgf/cm2) for 7 min revealed a maximum prediction saccharification rate of approximately 58.7% at 6.5% substrate concentration, 36.3 FPU enzyme concentration, and 330.3 mg/g-glucan surfactant (polysorbate 80) concentration. The extents of saccharification of Korean oak (99.9%) and pine (58.7%) demonstrated the high applicability of the crude enzyme from Acanthophysium sp. KMF001.