This study was conducted to evaluate the optimum dietary protein level in juvenile river puffer. Five semi-purified diets were formulated by using casein to contain graded levels of protein levels of 35, 45, 50, 55 and 65%. Fish averaging $8.56{pm}0.04g$ were randomly assigned to one of five experimental diets in triplicate groups for 8 weeks. After the 8-weeks of feeding trial, weight gain and feed efficiency of fish fed 45, 50 and 55% diets were significantly higher than those of fish fed 35 and 65% diets (P<0.05). Protein efficiency ratio of fish fed the 35% diet was significantly higher than those of fish fed 65% diet (P<0.05), but there were no significant difference among those of fish fed 45, 50 and 55% diets. Specific growth rate of fish fed 50% diet was significantly higher than those of fish fed 35 and 65% diets (P<0.05), but there was no significant difference among those of fish fed 45, 50 and 55% diets. No significant differences were observed in condition factor, hepatosomatic index, visceralsomatic index and survival among those of fish fed all the diets. Optimum dietary protein levels by using broken-line model and by using second order polynomial were estimated at 45.9% and 51.6% for the maximum growth of fish respectively. Therefore, these results suggested that the optimum dietary protein level could be greater than 45.9% but less than 51.6% for the maximum growth in juvenile river puffer.