Walnut kernel was used to research the influence of temperature and vacuum degree on its protein structure and function properties. The results showed that the quality of walnut kernel protein was significantly affected by factors of temperature and vacuum, and the influence degree of these factors was valued as: temperature > vacuum, constant temperature (35 ℃) > room temperature (25 ℃)> refrigeration (4 ℃). Moreover, with the extension of storage time, the content of sulfhydryl groups in walnut kernel protein decreased, and the content of disulfide bonds and carbonyl compounds increased. The results obtained from electrophoresis and infrared analyses indicated that the protein aggregation phenomenon occurred, and the ratio of β-sheet and random coils in the secondary structure increased, with the decrease of the ratio of α-helix to β-turn. In terms of functional properties, the surface hydrophobicity of walnut kernel protein increased, and solubility decreased, as well as its digestive properties significantly changed.