In order to provide theoretical guidance for processing all the year， the red swamp crawfish preboiled were placed in the plastic box filled with water， then the boxes with crawfish were vacuumed， followed that the crawfish were frozen in the ice locker at -20， -40， -55 ℃， respectively. After the center temperature of which was fallen to -15 ℃， the frozen crawfish in boxes was stored at -20， -40 ℃ for 24 weeks. The content， surface hydrophobicity， intrinsic fluorescence intensity， total sulfhydryl and disulfide bond contents， and secondary structure of myofibril protein as well as microstructure of ice crystal were measured， to investigate the effect of frozen and freezing storage temperatures on physicochemical properties of protein from red swamp crawfish. The results showed that， at the same freezing storage temperature， the ice crystal with red swamp crawfish frozen at -20 ℃ was bigger， resulting in a heavier damage of meat structure， compare to the frozen temperature with -40 ℃ and -55 ℃. Besides， the myofibril protein content of crayfish frozen at -20 ℃ was significantly lower than that of -40 ℃ and -55 ℃ (P<0.05)， while there is a slight difference in the surface hydrophobicity， intrinsic fluorescence intensity， and total sulfhydryl and disulfide bond contents of myofibril protein(P>0.05)； at the same frozen temperature， the ice crystals with freezing storage at -20 ℃ was larger， and the needle-shaped ice crystals appear earlier， compared to freezing storage temperature of -40 ℃. The myofibril protein and total sulfhydryl contents of crayfish with -20 ℃ significantly lower than that of -40 ℃ (P<0.05)， while the surface hydrophobicity and disulfide bond content were significantly higher than that of -40 ℃ (P<0.05). The intrinsic fluorescence intensity of myofibrillar protein with -20 ℃ changed faster than that of -40 ℃. In addition， the relative content of α-helix with frozen or freezing storage temperatures of -20 ℃ was higher than that of -40 ℃ and -55 ℃ frozen or -40 ℃ of freezing storage， while there is a slight difference in β-sheet， β-turn， and random coil. These results indicated that a lower frozen and freezing storage temperatures may keep a more stable spatial conformation of protein from red swamp crayfish and alleviate the degree of protein denaturation， resulting in better quality attributes of red swamp crayfish during freezing storage.