Abstract This study evaluated the impacts of replacing fish meal (FM) with defatted black soldier fly (Hermetia illucens; BSF) on the growth performance, flesh quality and transcriptome of Pacific white shrimp (Litopenaeus vannamei). In a diet containing 560 g/kg FM, BSF was used to replace 0%, 20%, 40%, 60%, 80% and 100% of dietary FM (BSF0, BSF20, BSF40, BSF60, BSF80 and BSF100). Thus, six isonitrogenous and isolipidic diets were prepared, and then fed to juvenile Pacific white shrimp (1.70 ± 0.10 g) for 60 days. In growth performance, the BSF20 group showed the similar FCR and WGR to the control group (), and the other four BSF groups presented significantly lower WGR and higher FCR than the BSF0 group (). The flesh chewiness and the contents of crude lipid, collagen and total amino acid in flesh were significantly decreased (), and flesh cooking loss, thawing loss was significantly increased when the replaced FM was ≥40% (). When the replaced FM reached 60%, the flesh hardness, heat-insoluble collagen content, shear force, body surface redness and the ratio of n-3/n-6 fatty acids were significantly lower (), while n-6 polyunsaturated fatty acid content and total free amino acid content in flesh were significantly higher than those of the BSF0 group (). Hepopancreatic samples were used for transcriptomic analysis, and a total of 1,456 differentially expressed genes (DEGs) were identified between the BSF groups and BSF0 group, which were mainly involved in growth-promoting, energy metabolism and antioxidant capacity key pathways and genes, including the cathepsin L, folate synthesis pathway, redox-related genes and glutathione metabolism pathway. In conclusion, in a diet containing 560 g/kg FM, BSF could successfully replace 20% of dietary FM, and higher FM replacement (≥40%) decreased the growth performance and flesh quality of Pacific white shrimp.