The study aimed to study the influence of different reaction temperatures on the carp (Cyprinus carpio) hepatic CYPs activity. Six groups of carp hepatic microsomes were incubated with probe drug (chorzoxazone) at 5, 10, 15, 20, 25 and 30°C separately. According to the principle of enzyme kinetics theory, the Michaelis constant (K (m)) value and maximum reaction velocity (V (max)) of CYPs (with CZX as probe) were obtained. The CYPs activity at different reaction temperatures was compared. In results, the K (m) values were separately 44.62, 31.35, 26.59, 21.75, 16.39, 29.69 μM, and the V (max) were separately 0.231, 0.234, 0.265, 0.294, 0.315, 0.239 nmol min(-1)mg(-1) at 5, 10, 15, 20, 25, 30°C. Results indicated that the enzyme CYPs activity was much higher at 25°C. It was also demonstrated that reaction temperature could affect the CYPs activity significantly. Therefore, this experiment builded a theoretical basis for the variations of fish pharmacokinetic parameters at different water temperatures and contributed to further research on the influence of water temperature on fish drug metabolism.