Abstract: White rot fungi preserved in our laboratory was used to produce manganese peroxidase.Design Expert 8.05 software was used to optimize the culture medium producing this peroxidase.3-Level factorial analysis was used to analyze the main effect factors from 10 factors.The result showed that the wheat bran, yeast extract and KH 2 PO 4 were the significant factors.According to Box-Behnken experimental design and response surface methodology, the second-order equation model was established by regression analysis of experimental data.The predicted optimal addition of wheat bran, yeast extract and KH 2 PO 4 were 10.75、3.37 and 0.095g/L respectively, and the predicted maximum manganese peroxidase could reach 4.06U/mL.The manganese peroxidase activity of verification experiment was 4.15U/mL, which was very close to the predicted value.The manganese peroxidase activity under the optimization condition increased by 58.4% compared with the original enzyme production medium.