This paper is concerned with optimization of submerged culture conditions for mycelial growth and bioactivity components production by Cordyceps militaris. Based on investigation of the most suitable carbon source and nitrogen source by one-factor-at-a-time method, the combination of carbon and nitrogen sources were optimized to investigate the effects on mycelial growth and bioactive components production using L9(34) orthogonal array. The effects of medium components on mycelial growth of C. militaris and adenosine production were in the order of soluble starch> dried silkworm chrysalis meal> fructose> soybean meal, and those on cordycepin production were in the order of fructose> soybean meal> dried silkworm chrysalis meal> soluble starch. The optimal medium combination, soluble starch 22.50 g/L, fructose 17.50 g/L, soybean meal 5.00 g/L and silkworm chrysalis meal 3.75 g/L, was beneficial for the accumulation of mycelial biomass (³2.36 g/L), while the optimal medium combination included soluble starch 12.50 g/L, fructose 22.50 g/L, soybean meal 3.75 g/L and silkworm chrysalis meal 3.75 g/L for the accumulation of adenosine (³193.67 µg/g), and soluble starch 12.50 g/L, fructose 17.50 g/L, soybean meal 3.75 g/L and silkworm chrysalis meal 5.00 g/L for the accumulation of cordycepin (³243.33 µg/g).