Abstract: The fruit bodies of Tremella aurantialba in Yunnan were used as materials to obtain the optimum method of Tremella aurantialba polysaccharide aqueous enzymatic extraction. The polysaccharide extraction yield was taken as evaluation index, enzyme dosage, liquid-solid ratio, extraction temperature and extraction time were investigated through the response surface methodology design based on single factor method combined with Box-Behnken central composite method. Results showed that, the optimal parameters were determined as follows:Enzyme dosage was 20.50 mg/g, liquid-solid ratio was 347:1 (mL/g), extraction temperature was 52℃ and extraction time was 52 min. Under these conditions, the maximum extraction yield of Tremella aurantialba polysaccharide was 12.69%±0.52%. Compared with traditional method, the optimum aqueous enzymatic extraction of polysaccharide from Tremella aurantialba was obtained, which had lower extraction temperature and less extraction time but higher extraction yield.