This study aimed to evaluate the effects of certain grain medium conditions, such as carbon and nitrogen source addition, water content, and initial pH, on the hispidin production of Phellinus linteus using solid-state fermentation. The results showed that the highest hispidin yield, specific productivity of hispidin, and total content of hispidin were associated with sucrose and malt extract addition, 55% water content, and an initial pH of 5.5, which resulted in 0.290, 0.233, 0.301, and 0.296 mg/g dry weight of mycelium; 0.261, 0.191, 0.257, and 0.227 mu g/mg week(-1) specific productivity; and 46.01, 40.67, 58.85, and 55.06 mg/kg dry weight brown rice medium, respectively. Additionally, two combinations of culture conditions with pearl barley and black rice medium were tested for hispidin production. The highest hispidin yield, specific productivity of hispidin, and total hispidin content for pearl barley medium fermented using P. linteus were 3.88-fold, 4.86-fold, and 3.60-fold higher than those for the control (brown rice medium), respectively. Overall, this study shows that P. linteus hispidin production can be enhanced using solid-state fermentation, with optimal medium conditions. (C) 2020, The Society for Biotechnology, Japan. All rights reserved.