Abstract: The effect of carbon resource, nitrogen source, Cu2+ and other factors on laccase secretion by and cell growth of Flammulina velutipes LP03 were investigated. The optimum conditions were 3 g/100 mL corn flour, 1 g/100 mL peptone，0.5 g/100 mL glucose，1 mmol/L copper sulfate, initial pH 6.0, 200 mL/500 mL liquid medium, 28 ℃ and 160 r/min. The maximal enzyme activity was (1813.52 ±5.59) U/L, which was 3.78 times of that before optimization.

Key words: Flammulina velutipes, laccase, culture condition