Abstract: In this report, response surface methodology was applied to optimize the fermentation conditions Coriolus versicolor ZZH-2 for Laccase Production. Firstly, Plackett-Burman design was used to select the main factors affecting laccase yield of Coriolus versicolor ZZH-2. The results showed that three factors played important roles in the fermentation conditions, including the concentration of yeast extract, sodium carboxymethyl cellulose (CMC) and L-aspartic acid. The pass of steepest ascent was undertaken to approach the optimal region of the three significant to factors. Then the central composite design and response surface analysis were applied to investigate the mutual interaction between the variables and identify optimal values that bring maximum laccase activity. The activity of laccase increased nearly 1.7 times to 9693U/mL where concentrations of yeast extract, sodium carboxymethyl cellulose (CMC) and L-aspartic acid were at 13.82, 13.29 and 1.44g/L. The experimental values agreed with the predicted values.