Abstract: Kombucha beverage of symbiosis fermentation by immobilized cell technology had an important significance for its industrial production. In this experiment, the total acid content, total sugar utilization and sensory evaluation results of the fermentation broth was used as index. In the first, the study discussed the effects of fermentation conditions including the amount of sucrose added, the inoculum rate, the ratio of strain inoculum and the fermentation time on symbiosis fermentation of Kombucha beverage by immobilized cell technology. And then orthogonal experiment was used to optimize the preparation process of Kombucha beverage of symbiosis fermentation by immobilized cell technology. The results showed that the optimal condition of symbiosis fermentation of Kombucha beverage by immobilized cell technology for:the amount of sucrose added was 70g/L, the inoculum rate was 7.5%, the ratio of strain inoculum was 6∶4, fermentation lasted 6 days.The Kombucha beverage produced according to this preparation technology remained uniform color, pleasant aroma, the distinctive flavor of mellow sweet and sour.