Cordyceps militaris was one of the most popular edible mushrooms. It is a nutritious food source and contains bioactive metabolites with several medicinal effects including anti-tumor, hypocholesterolemic, antioxidant and anti-diabetic activities. In this study, fibrin fibrinolytic enzyme-producing strains were screened from Cordyceps militaris by plate method and specific activity. The fermentation conditions of the fibrinolytic enzyme producing were investigated. The results showed that the best fermentation medium contained 2% glucose, 2% peptone and 1/1 the ratios of carbon to nitrogen. The optimum fermentation conditions were pH 6.0, the temperature 25 ℃, the rotation speed 150 r/min, the fermentation time 9 d, the inoculum size 2% and medium volume 60 mL. Under such conditions, the fibrinolytic enzyme activity was 111.85 U/mL.