Nutrition intervention is an effective way to improve flesh qualities of fish. The effect of feed supplementation with glutamate (Glu) on flesh quality of gibel carp () was investigated. In trial 1, the fish (initial weight: 37.49 ± 0.08 g) were fed two practical diets with 0 and 2% Glu supplementation. In trial 2, the fish (37.26 ± 0.04 g) were fed two purified diets with 0 and 3% Glu supplementation. The results after feeding trials showed that dietary Glu supplementation increased the hardness and springiness of muscle, whether using practical or purified diets. Glu-supplemented diets increased the thickness and density of myofibres and collagen content between myofibres. Furthermore, Glu promoted muscle protein deposition by regulating the IGF-1-AKT-mTOR signalling pathway, and enhanced the myofibre hypertrophy by upregulating genes related to myofibre growth and development (, , , , , and ). The protein deposition and myofibre hypertrophy in turn improved the flesh texture. In addition, IMP content in flesh increased when supplementing Glu whether to practical or to purified diet. Metabolomics confirmed that Glu promoted the deposition of muscle-flavoured substances and purine metabolic pathway most functioned, echoed by the upregulation of key genes (, and ) in purine metabolism. The sensory test also clarified that dietary Glu improved the flesh quality by enhancing the muscle texture and flavour. Conclusively, dietary Glu supplementation can improve the flesh quality in this fish, which can further support evidence from other studies more generally that improve flesh quality of cultured fish.