In comparison with conventional thawing methods (water immersion thawing (WIT), natural air thawing (NAT), and low-temperature air thawing (LAT)), the influences of low temperature combined with high-humidity thawing (LT)(-1-1℃, 2-4℃, 5-7℃ and 8-10℃, RH≥95%) on the quality characteristics of Penaeus vannamei were evaluated, including thawing time, water holding capacity, pH, color, texture, lipid oxidation, and biochemical properties of myofibrillar protein. The results showed that WIT was fastest (only costing 28.40min) and LAT was slowest (costing 367.33min) thawing methods, respectively. High humidity was beneficial to thaw, and the thawing rate at 2-4℃ was 9.14% and 57.62% higher than that of NAT and LAT, respectively. LT treatment at -1-7℃ could maintain the water holding capacity and texture quality of Penaeus vannamei well. The minimum thawing juice loss of LT treatment was observed at 2-4℃, which was only 16.32% to 23.40% of that in three conventional thawing methods, and the centrifugal loss was only 2.80%. The best texture quality was exhibited at 2-4℃. And the hardness and chewiness were significantly higher than those of conventional thawed samples. Oxidation of lipid and myofibrillar protein showed an apparent decrease with LT treatment (-1-7℃). Compared with that in conventional thawed samples, the levels of thiobarbituric acid reacting substances were remarkably decreased, while total sulfhydryl contents were dramatically increased. The surface hydrophobicity of LT treated samples at 2-4℃ was even closed to the fresh level, which was only 7.69μg/mg (based on per milligram protein). Moreover, the activity of Ca²⁺-ATPase increased obviously under LT treatment (-1-7℃), indicating the denaturation of protein was lower than that of conventional thawed samples. Considering the thawing efficiency, product quality and production practice, LT was an ideal method for the thawing of Penaeus vannamei.