Methyl orange wastewater was treated by domesticated Trichoderma koningii. Taking the decolorization rate as the index, the effects of culture factors such as reaction time, initial methyl orange concentration, pH, glucose dosage and different culture time on the degradation of methyl orange wastewater in the early growth stage of Trichoderma koningii were discussed. Taking the decolorization rate as the index, the enzymatic degradation reaction under the condition of certain pH, temperature, enzyme solution dosage, initial methyl orange concentration and reaction time was discussed; the optimum conditions for decolorization and degradation of methyl orange dye were verified by orthogonal test, which provided an experimental basis for the biological treatment of dye wastewater. The results showed that the optimum condition for the degradation of methyl orange wastewater by Trichoderma koningii was initial methyl orange concentration of 30 mg/L, culture time of 72 h, glucose concentration of 25 g/L and pH of 6.5. The degradation rate of the enzyme extracted under the optimal culture condition can reach 85.48% under the treatment condition of enzyme solution dosage of 0.6 mL, temperature of 25℃, pH of 7.0 and treatment time of 6 h, the decolorization effect is obvious, which provides a reference for the enzymatic degradation condition of methyl orange dye wastewater by Trichoderma koningii.