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BACKGROUND The canned-sardine industry generates large amounts of protein-rich waste, which demands useful exploitation. This paper describes the potential use of muscle and viscera proteins from canned sardine by-products as substrate to obtain hydrolysates with biological and functional properties. Three enzymatic approaches, brewer's spent yeast (Bsy) proteases, Alcalase® and Neutrase® were applied to perform protein hydrolysis at the same proteolytic activity (1 U mL−1), using an enzyme/substrate ratio of 20% (v/v), at 50°C and for 7 h. Hydrolysis degree (DH), antioxidant and angiotensin I-converting enzyme inhibitory (ACE-I) activities, functional properties (i.e. solubility, emulsifying and foaming properties, water and oil binding capacity) and colour were investigated. RESULTS All hydrolysates presented a high protein content [52.7-83.2% dry weight (DW)] and low fat content (0.9-3.9% DW). Alcalase® treatment of muscle and viscera proteins resulted in higher DH (7.5% and 8.6%, respectively) and higher biological activities ( P < 0.05). All hydrolysates had excellent solubility and presented functional properties. Among viscera hydrolysates, treatment with Bsy proteases promoted higher emulsion (80.1 m2 g−1), foaming (79.2%) and oil binding capacity (5.8 g g−1) of viscera sardine proteins. CONCLUSION Improved biological and functional properties were observed for sardine protein hydrolysates produced using the three enzymatic treatments tested. © 2017 Society of Chemical Industry [ABSTRACT FROM AUTHOR]