In order to improve the activity of laccase by Trametes versicolor FG-97, response surface methodology was applied to optimize the fermentation conditions. We first optimized the conditions by using Plackett-Burman design to screen for the significant factors, found that pH, temperature and incubation time were the main ones. After the steepest ascent test and Box-Behnken design, the optimum conditions for laccase production were found to be a temperature of 30 °C, pH of 5.2 , time of fermentation for 9 days (160r/min) and 2% (w/v) of inoculation density, the peak of laccase activity achieved at 306.33 U·L-1 ，and the experimental values agreed with the predicted values. These results suggested that the predicted model was reliable and available for the optimization of laccase fermentation conditions.