Proteinase (endopeptidase), trypsin and chymotrypsin activities, RNA, and DNA contents were examined throughout the ontogenetic development of cultured Litopenaeus schmitti. Whole individuals from larval and postlarval stages, and adult hepatopancreas were homogenized and assayed for quantification of enzyme activity. Proteinase activity of different life stages was characterized by substrate-SDS-PAGE. Specific inhibitors for trypsin (TLCK) and serine proteinases (PMSF) were used to identify activity zones of these enzymes in gels. Nucleic acids were also quantified in larval and postlarval stages. Protein-specific activity of total proteinases and trypsin presented higher values in nauplius IV (N IV), protozoea I (PZ I), and PZ III, compared to the remaining stages, while chymotrypsin activity peaked in N IV and PZ III. Enzyme activity was lower in egg, early nauplius, mysis, and postlarval stages. Different proteinase patterns were observed in SDS-PAGE during ontogenetic development. Active bands of 15.5, 16.0, 19.9, 21.8, 24.0, 27.9, 30.3, 33.5, 35.5, 38.2, and 48.1 kDa were detected in the adult hepatopancreas. In the course of development, proteolytic activity was detected in N IV, and intense bands of 17.3, 19.9, and 20.9 kDa were found up to mysis (M III). A band at 38.2 kDa was present between N IV and M I, and between PL II and PL IV. Most bands were serine proteinases, and only two bands in adult (15.5 and 15.9 kDa) were inhibited by TLCK. Both RNA and DNA contents (μg mg −1 of fresh weight) reduced with development from egg to PZ III, increasing afterwards until PL IV. RNA/DNA ratios increased from egg, reaching a peak in PZ I, and decreased in the following stages until a minimum was reached in M II. A secondary peak was observed in PL III followed by reduction in PL IV. The variation in enzyme activity indicates different strategies of energy use throughout development. Higher enzyme activity combined with increased RNA/DNA in protozoeal stages denotes intense food energy use and accumulation into growth. The shift in energy strategy during the ontogeny of L. schmitti is accompanied by variation in proteinase characteristics as observed in other penaeid species.