Auxistapeionosoma Bleeker is a kind of tuna in the South China Sea with high yield and low value. This study analyzed the proximate composition, amino acid composition of the ordinary meat and dark meat hydrolysates, and evaluated their anti-fatigue activity by animal experiment. The total amino acids of the ordinary muscle hydrolysate powder (OMHP) and dark muscle hydrolysate powder (DMHP) of Auxistapeionosoma Bleeker were 57.06% and 58.90%, respectively. The branched chain amino acids accounted for about 18% of the total amino acids. The contents of the components in OMHP and DMHP with molecular weight below 5000Da were both up to 70%, and the content of below 700Da was both more than 20%.Compared with the control group, the OMHP and DMHP could significantly prolong the swimming time of mice (p<0.05), increase liver glycogen reserves of mice (p<0.05), and lower the serum urea nitrogen in mice after exercise (p<0.05). It showed that both had anti-fatigue activity, and there was no significant differences between them. The results showed that the OMHP and DMHP of Auxistapeionosoma Bleeker from South China Sea could be used as potential anti-fatigue health care products.