This study assessed the effect of protein level in diet and feeding schedule on the digestive enzymatic activity of Macrobrachium tenellum juveniles. Freshwater prawn juveniles (0.31 ± 0.004 g and 32.48 ± 0.27 mm) were fed diets with 20, 25, 30, 35 and 40% crude protein (CP) (p/p). In a second experiment, feeding schedules were assessed at 08:00, 14:00 and 20:00 h with feed at 30% CP with juveniles of 0.22 ± 0.002 g and 29.36 ± 0.28 mm. In both experiments, five organisms were sampled after 60 days of treatment in a time-lapse of one hour (08:00 to 09:00 h) per experimental tank and treatment. The enzymatic extract was obtained from the complete organisms to determine specific general protease, trypsin, chymotrypsin, amylase and lipase activities. Greater general protease, trypsin and lipase activities were observed in feed at 40% CP. The feeding schedules with greater general protease, trypsin, lipase and amylase activities were found at 14:00 and 20:00 h. Chymotrypsin did not show differences among treatments at CP levels or feeding schedules. Protein level in diets for M. tenellum modified basal digestive enzymatic activity and increased proteolytic and lipolytic activities, while it decreased amylolytic activity as the inclusion of CP level increased in the diet; likewise, the activity was modified by feeding schedule, showing a direct correlation between the increase in digestive enzymatic activity and food offering at the end of the day.