An 8‐week feeding trial was conducted to estimate the optimum dietary manganese (Mn) requirement for juvenile hybrid grouper, Epinephelus lanceolatus × E. fuscoguttatus. The basal diet was formulated to contain 520 g/kg crude protein from casein and fishmeal. Manganese methionine was added to the basal diet at 0 (control group), 2.5, 5, 10, 20 and 40 mg Mn/kg diet providing 7.48, 10.34, 13.76, 19.72, 31.00 and 53.91 mg Mn/kg diet, respectively. Each diet was randomly fed to triplicate groups of juveniles, and each tank was stocked with 20 fish (initial weight, 60.06 ± 0.68 g). The manganese content in rearing water was monitored and kept below 0.01 mg/L. Results showed that the weight gain ratio (WGR), protein efficiency ratio (PER), specific growth rate (SGR), Mn contents in whole body, liver and vertebra, and activities of hepatic Mn superoxide dismutase (Mn‐SOD), total SOD (T‐SOD) and glutathione peroxidase (GSH‐PX) were significantly improved by dietary Mn supplementation (p < .05). However, dietary Mn did not affect arginase (DArg) activity. The highest feed conversion ratio (FCR) was observed in fish fed the basal diet (p < .05). No significant differences were found on the Cu and Zn contents in whole body by supplementing dietary Mn. Supplemented Mn in diets had significantly effect on liver and vertebral trace element deposition (p < .05). Fish fed the basal diet had the highest Fe and Zn contents in vertebra (p < .05). There were no significant differences on hepatic pyruvate decarboxylase (PDC) activity with supplemented Mn levels below 13.76 mg/kg. As biomarker of oxidative stress, malondialdehyde (MDA) content in liver was significantly higher in fish fed the basal diet (p < .05). Using the broken‐line models based on SGR, dietary Mn requirement of the juvenile hybrid grouper was estimated to be 12.70 mg/kg diet.