Abstract: To investigate the effects of dietary protein levels on growth, feed utilization, and energy metabolism of Pacific white shrimp (Litopenaeus vannamei) under high-temperature conditions. Three practical diets containing 40%, 42.5%, and 45% crude protein were formulated and designated as the P40, P42.5, and P45 groups, respectively. Juvenile Pacific white shrimp with an initial body weight of (5.42±0.01) g were cultured at 32℃ for an 8-week feeding trial. The results showed that the weight gain rate and specific growth rate of shrimp in the P42.5 group were significantly higher than those in the P40 and P45 groups, while the feed conversion ratio was significantly decreased (P<0.05). Hemolymph biochemical analysis indicated that the P42.5 group had the highest albumin content and superoxide dismutase (SOD) activity, whereas the P45 group exhibited the highest hemolymph ammonia level. For hepatopancreatic antioxidant indices, the activities of lysozyme (LZM), catalase (CAT), and other related enzymes reached their maximum values in the P42.5 group. Compared with the other two groups, malondialdehyde (MDA) content in the P45 group was significantly increased (P<0.05). Regarding intestinal health, the P42.5 group showed the highest intestinal villus height and mRNA expression levels of tight junction protein genes (Claudin-3, Occludin and ZO-1). In terms of protein synthesis, the transcript levels of translation initiation factor genes (eIF3K and eIF4E) were the highest in P42.5 group and the lowest in P40 group. For energy metabolism, the expression levels of key enzyme genes related to the TCA cycle (IDH, MDH, and CS) and the concentrations of corresponding metabolites peaked in the P45 group. Under the rearing temperature of 32℃, a dietary protein level of 42.5% effectively promoted shrimp growth, enhanced antioxidant capacity and intestinal barrier function, and maintained the homeostasis of protein synthesis and energy metabolism. A dietary protein level of 40% suppressed energy metabolism and resulted in reduced growth performance. In contrast, excessive dietary protein (45%) aggravated oxidative stress, impaired intestinal barrier integrity and led to the decline of growth performance in Pacific white shrimp. Therefore, reasonable regulation of dietary protein supply is an important approach to improve the culture stability, nutrient utilization efficiency and health status of Pacific white shrimp under high temperature conditions.