Abstract In order to optimize the Phellinus Linteus fermentation process, the fermentation temperature, the inoculum size, the rotation speed and the bottling capacity have been regarded as the independent variables and the fermentation yield has been considered as the dependent variable. The quadratic regression orthogonal rotating combination design method has been adopted to build the model for Phellinus Linteus fermentation process. And this model has been also analyzed in this paper. The results have shown that the most important factors which affect the fermentation yield include the fermentation temperature, the inoculum size, the rotation speed and the bottling volume. There exists the significant interaction between the inoculum size and the bottling volume. The optimal fermentation conditions of Phellinus Linteus are: the bottling volume is 120mL, the inoculum size is 17mL, the temperature is 26°C and the rotation speed is 135r/min. At this time, the theoretical extreme value of fermentation mycelium production is 24.51mg/mL. The experimental results have shown that the model fits well with the actual situation and the credibility is higher.