The present study was designed to determine the optimal dietary lysine requirement for jundiá, Rhamdia quelen, fingerlings. Groups of 17 fish (1.4 ± 0.1 g) were stocked in 120‐L tanks and were fed semipurified diets (33% crude protein [CP] and 3500 kcal metabolizable energy) containing increasing concentrations of lysine (3.0, 4.0, 4.5, 5.0, 5.5, 6.0, and 6.5% CP). After 119 d, fish weight gain (WG), specific growth rate (SGR), feed intake and feed conversion (FC), apparent net protein utilization (ANPU), body composition (CP, fat, and ash), and vertebral collagen were determined. WG and SGR increased as dietary lysine concentration in protein increased up to 4.5%, reducing at 6.0 and 6.5% lysine. Fish that were fed the lowest lysine concentration presented the worst feed conversion (FC), which improved for fish fed with 4.5% or more lysine. Feed consumption followed the same trend as FC. The highest ANPU was observed in fish fed with 4.5% lysine. Fish fed diets containing 4.5, 5.0, and 5.5% lysine accumulated more body protein (P < 0.05). Collagen vertebral concentration was significantly higher in fish fed with the 4.5% lysine diet. Dietary requirement for lysine was 4.5 or 5.1% depending on the statistical model used for estimation: broken line or polynomial regression, respectively. The requirements for the other essential amino acids were estimated on the basis of the ideal protein concept and were similar to the requirements for other fish species, except for isoleucine, leucine, treonine, and valine, which were higher for jundiá.