In order to evaluate the effect of novel cryoprotectants composed of pectin, collagen peptide and trehalose on the quality of quick-frozen pumpkin puree and relative mechanism, the stability of the pumpkin puree was characterized by the water separation proportion, average particle size and Zeta potential value, and the product quality was evaluated by texture properties, color and total soluble solid content. Scanning electron microscope (SEM) and laser scanning confocal microscope (CLSM) were also used to observe the microstructure of the sample. The results showed that the cryoprotectants could significantly slow down the increase tendency in WSP and particle size, and also prevent the decrease in the L*, a*, b* value, hardness, cohesiveness, total soluble solid content and Zeta potential value. The observation result of SEM and CLSM showed that cryoprotectants could effectively protect the microstructure of quick-frozen pumpkin puree and reduce the degree of particle aggregation. The findings of this study provide theoretical guidance for the development of pumpkin puree frozen prepared food.