Protein hydrolysates have been widely reported as the protein source of aquatic feed. However, previous studies on protein hydrolysates focused on fish under experimental conditions. In this study, a 6-week feeding trial in a greenhouse was conducted to investigate the effects of partially replacing fishmeal by protein hydrolysates on growth performance, digestive enzymes, and TOR signaling pathway of Litopenaeus vannamei under practical conditions. This involved randomly selecting 72,000 shrimps (initial body weight 2.26 ± 0.02 g) and placing them in groups inside nine concrete tanks (L 5 m × W 5 m × H 1 m) (3 treatments × 3 replicates × 8000 individuals per concrete tank). Two isonitrogenous (430 g kg-1) and isolipidic (80 g kg-1) diets were prepared: fishmeal diet (FM) containing 400g kg-1 fishmeal, and protein hydrolysates diet (PH) in which 15% of the fishmeal was replaced by protein hydrolysates. A commercial diet (CD) was used as reference. The final weight (FW), percent weight gain (PWG), specific growth ratio (SGR), and total weight for each tank (TW) of L. vannamei fed with FM and PH diets were not significantly different (P > 0.05). However, shrimp fed with PH diet had significantly higher FW, PWG, SGR, and TW values than those fed with CD diet (P < 0.05). Trypsin activity was significantly lower in shrimp fed with CD diet, than in shrimp fed with FM and PH diets (P < 0.05). However, trypsin activity of L. vannamei fed with FM and PH diets were not significantly different (P > 0.05). The mRNA expression of tor, s6k, and 4e-bp genes were not significantly affected between FM and PH diets, while tor and s6k expression levels of CD diet were significantly down-regulated. Based on the above data, 15% replacement of fishmeal with protein hydrolysates did not make any difference on shrimps compared with FM group. Therefore, protein hydrolysates can partially replace fishmeal as the protein source of shrimp formula feed in practical conditions.