The objectives of the study were to screen the strain with a good antioxidation activity from 10 species of wild macrofungi and optimize the optimal conditions for antioxidant substance production. These results can provide some references for the breeding of Auricuiaria auricular and industrialization of its production. As material with 10 strains of wild fungi from the Langyashan in Anhui province, the strains with antioxidant substance producing were obtained by DPPH free radical scavenging rate of the broth. Shake flask culture was then used to get the optimum culture conditions based on the single factor and orthogonal experiments, which is finally examined. The results showed that there were four strains with producing antioxidation activity in the samples, accounting for 40%. The strain of Auricuiaria auricular has especially the higher capacity of antioxidaton activity. The formula of appropriated culture conditions were as follows: glucose 25 g/L, yeast extract 2 g/L, MgSO₄ 0.2 g/L, CuSO₄ 6 mg/L, K₂HPO₄ 3.2 g/L, pH 6.0, 27 ℃, culture volume 100 mL/250 mL, and cultivation 8 d. The metabolites of the Auricuiaria auricular have good antioxidant activity, providing a new source of natural antioxidants development.