To investigate the impact of varying protein levels in feed on the growth performance, ovarian amino acid and fatty acid composition, gonad development, and serum hormone deposition of pengze crucian carp (Carassius auratus var. Pengze) broodstock, 135 broodstock with an average initial weight of (256.30±4.70) g were randomly selected for the experiment. These fish were then divided into three groups, each with three replicates and 15 individuals in each replicate. Fish and soybean meals were the primary protein sources, while corn starch was used to adjust the protein gradient, with dietary vitamin E content at 0 mg/kg (68.1 mg/kg), 400 mg/kg (462.3 mg/kg), and 1600 mg/kg (1655.8 mg/kg). The experiment was conducted for 60 days. No significant difference was observed in the growth, survival rate, and weight gain rate of Pengze crucian carp (P＞0.05). The fattening degree showed a significant increase and a stable trend with the increase in vitamin E in the feed. The gonadal index of the 400 mg/kg group was significantly higher than that of the remaining two groups (P＜0.05), and the egg diameter and number of eggs conceived showed an increasing trend and a decrease with an increase in vitamin E (P＞0.05). In the liver, vitamin E content in the 1600 mg/kg group was significantly higher than that in the remaining two groups (P＜0.05). In the ovaries, the vitamin E content in the 400 and 1600 mg/kg groups was significantly higher than that in the 0 mg/kg group (P＜0.05). The addition of vitamin E significantly increased the total amount of essential amino acids in the ovary (P＜0.05). Valine, isoleucine, threonine, lysine, and the total essential amino acids in the 400 mg/kg group were significantly higher than those in the 1600 mg/kg group (P＜0.05). Histidine, glutamic acid, and total amino acids were significantly higher in the 1600 mg/kg group than in the remaining two groups (P＜0.05); however, no significant difference was observed in the total non-essential amino acids between the groups (P＞0.05); seven saturated fatty acids, four monounsaturated fatty acids, and eight polyunsaturated fatty acids were detected in the ovaries of the various groups (P＜0.05). Polyunsaturated fatty acids, C18:3n6, and ΣPUFA were significantly lower in the 1600 mg/kg group than in the remaining two groups (P＜0.05), whereas DHA and Σn-3PUFA peaked in the 400 mg/kg group and were significantly higher than those in the 0 mg/kg group (P＜0.05), but did not differ significantly from the 1600 mg/kg group (P＞0.05). In the serum, CAT, ACP, and VTG were significantly higher in the 400 and 1600 mg/kg groups than in the 0 mg/kg group (P＜0.05), and MDA was significantly lower (P＜0.05), while T-AOC and LZM reached their peaks in the 400 mg/kg group and were significantly higher than that in the 0 mg/kg group (P＜0.05). In the liver, the SOD activity peaked in the 400 mg/kg group and was significantly higher than that in the 0 mg/kg (P＜0.05), and MDA activity decreased significantly with increasing feed vitamin E levels (P＜0.05). In conclusion, the addition of 400 mg/kg vitamin E in the diet significantly increased the gonad index and vitamin E content in the ovary of Pengze crucian carp, which was beneficial to the deposition of amino acids and fatty acids and increased the immune and antioxidant enzyme activities in serum and liver, thereby improving the gonadal development of Pengze crucian carp.