Using Grifola frondosa and crabapple as the raw materials, Lactobacillus plantarum MA2, Lactobacillus bulgaricus 717 and Streptococcus thermophiles 176 as the fermentation starters， a fermented G. frondosa crabapple beverage was prepared, and the fermentation process was optimized by orthogonal experiments. The results showed that the optimum process technology for fermented G. frondosa crabapple beverage was as follows: granulated sugar addition 6%, starters MA2∶717∶176 inoculation ratio 2∶1∶1, fermentation time 3 d, and temperature 37 ℃. Under the optimized condition, the beverage was sour and sweet refreshing with good flavor, and the sensory score was 89. The anti-fatigue effect of the beverage on rats was explored through cage running, weight-bearing swimming and pole climbing experiments. The results showed that the fermented beverage had good anti- fatigue effect, which significantly improved the body endurance of rats and alleviated fatigue.