Abstract: OBJECTIVE To increase the yield of fungichromin(FC) in actinomycetes strain WP-1 through optimizing fermentation conditions. METHODS The fermentation medium components were optimized by single factor and orthogonal experiments to investigate the effect of different available or slow-acting carbon sources, nitrogen sources and different inorganic salts on the yield of FC. The culture conditions of actinomycetes strain WP-1 were optimized by single factor experiments to investigate the effect of different initial pH values of medium, culture temperatures, inoculation amount, fermentation volume and rotational speeds on the yield of FC. RESULTS The optimization results were as follows:glucose 1%, soluble starch 4%, peptone 0.5%, peanut powdery cake 9%, MnSO₄ 0.000 5%, ZnSO₄ 0.000 5%, CuSO₄ 0.000 2%; initial pH 5.5, T 28℃ loaded liquid 50 mL medium/250 mL triangular flask, inoculum size 8%, rotate speed 180 r min^-1. Under the optimized fermentation conditions, FC yield of actinomycetes strain WP-1 were 159.0 mg·L^-1, about 4.9 times of that under the initial fermentation conditions. CONCLUSION The best fermentation condition to produce FC is obtained so that this paved the way for next step of large scale fermentation.