l‐carnitine (LC) is required for transporting long‐chain fatty acids into the mitochondria, where β‐oxidation takes place, and it works as an antioxidant molecule against reactive oxygen species. This study evaluated the effects of LC on the growth and antioxidant function of Amur minnow (Phoxinus lagowskii Dybowskii). Five isonitrogenous (380.4 g/kg) and isoenergetic (17.63 MJ/kg) diets were supplemented with five LC levels: control level (0 mg/kg) and treatment levels (50, 400, 750, or 1,100 mg/kg) were fed to fish (18.19 ± 0.56 g) for 120 days. The results showed that the growth performance of fish fed a diet containing 400 mg/kg of LC was significantly higher than that of the control and those fed other LC level treatments. Similarly, the 400 mg/kg treatment had the best feed efficiency. Further, the levels of total antioxidant capacity and total glutathione in the serum and hepatopancreas of fish fed a diet containing 750 mg/kg of LC were significantly increased; however, malondialdehyde levels were significantly reduced compared to those of the control group. The activities of antioxidant enzymes of 750 mg/kg treatments in the serum and hepatopancreas were significantly higher than those of the control group, including total superoxide dismutase, catalase, glutathione peroxidase and gamma‐glutamyl‐cysteine synthetase. Finally, 750 mg/kg treatment significantly upregulated the mRNA relative expression of antioxidant enzymes and nuclear factor erythroid‐2‐related factor 2 and inhibited the mRNA level of kelch‐like ECH‐associated protein 1 in the hepatopancreas. In conclusion, the dietary LC level of 400–750 mg/kg could improve the growth performance, feed utilization and antioxidant defense system of Amur minnow under the culture conditions.