This study aims to develop a spore fermentation method instead of fruiting body extraction for the production of Tremella aurantialba polysaccharide (TAPS). The purified spore strain Tremella aurantialba NX-20 was isolated for TAPS fermentation. The fermented TAPS and the extracted TAPS had the same monosaccharide composition but higher molecular weight. The conditions for TAPS fermentation by NX-20 were optimized, which were 25 °C, pH 7, 10% initial inoculum and 5 days. To further reduce the processing steps and costs of the medium, tofu wastewater (TW) was used to replace defatted soybean meal (DSM). In a 7.5 L fermentation tank, 40 g/L glucose with 10-fold diluted TW was used as the medium. For TAPS fermentation, a maximum yield of 15.02 ± 0.40 g/L was achieved. In conclusion, this study provides a feasible strategy for the efficient preparation of TAPS through spore fermentation.