This study was conducted to estimate the optimum dietary protein to energy (P/E) ratio in juvenile river puffer. Nine experimental diets were formulated with three energy levels and three protein levels at each energy levels. Three energy levels of 3000, 3750 and 4500 kcal diets were included at 45, 50 and 55% crude protein (CP) levels, respectively (45P3000, 45P3750, 45P4500, 50P3000, 50P3750, 50P4500, 55P3000, 55P3750 and 55P4500). Fish averaging $3.43{pm}0.02$ g randomly were fed the experimental diets in triplicate groups for 8 weeks. Weight gain of fish fed the 50P4500 diet were significantly higher than that of fish fed the 45P3000, 45P3750, 45P4500, 50P3000 and 50P3750 diets (P<0.05), but there was not significantly different from that of fish fed the 50P4500, 55P3750 and 55P4500 diets. Feed efficiency of fish fed the 50P4500 diet were significantly higher than that of fish fed the 45P3000, 45P3750, 45P4500, 50P3000 and 50P3750 diets (P<0.05), but there was not significantly different from that of fish fed the 50P4500, 55P3000, 55P3750 and 55P4500 diets. Protein efficiency ratio of fish fed the 45P3000 and 45P3750 diets was higher than that of fish fed 50P4500 and 55P4500, but there was not significantly different from that of fish fed the 45P3000, 45P3750, 45P4500, 50P3000, 50P3750, 55P3000 and 55P3750 diets. Based on weight gain, feed efficiency and specific growth rate, diets containing energy levels 4500 kcal/kg diet had an optimum P/E ratio of approximately 111 mg protein/kcal (50% crude protein) in juvenile river puffer.