Antioxidant enzymes, scavengers of the reactive oxygen intermediate (ROI), are involved in numerous defense systems in cells. In the present study, we investigated the effects of the hot-water extracts of two medicinally potent mushrooms (Ganoderma lucidum and Phellinus linteus) on the activity and expression of antioxidant enzymes in vitro and in vivo. The mushroom extracts stimulated the catalase activity in a dose-dependent manner in vitro, whereas the other antioxidant enzymes (such as superoxide dismutase (SOD), glutathione peroxidase (GPx)) were unaffected by the extracts. The catalytic activity of catalase in the liver and brain was significantly increased after the oral treatment of the mushroom extracts (2.5 g/kg) to ICR mice for 2 months. Western blot analysis of the liver and brain tissues revealed that the expression level of catalase in the mice, treated with both mushroom extracts, was significantly increased compared to that of the control mice. However, the level of the SOD expression in the mice treated with the natural product extracts was unchanged under the same experimental conditions. Although the mechanisms for the stimulatory effect of the catalase expression by these extracts remains unclear, these results suggest that the ingredients of the Ganoderma lucidum and Phellinus linteus extracts act as an activator of catalase, and regulate the expression of catalase at the translational or transcriptional level.