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Boarfish (Capros aper) protein hydrolysate has potent insulinotropic and GLP‐1 secretory activity in vitro and acute glucose lowering effects in miceThe anti‐diabetic actions of a boarfish protein hydrolysate (BPH) were investigated in cultured cells and mice. A boarfish (Capros aper) muscle protein hydrolysate was generated using the enzymes Alcalase 2.4 L and Flavourzyme 500 L. Furthermore, the BPH was subjected to simulated gastrointestinal digestion (SGID). BPH and SGID samples (0.01–2.5 mg mL−1) were tested in vitro for DPP‐IV inhibition and insulin and GLP‐1 secretory activity from BRIN‐BD11 and GLUTag cells, respectively. The BPH and SGID samples, caused a dose‐dependent increase (4.2 to 5.3‐fold, P < 0.001) in insulin secretion from BRIN‐BD11 cells and inhibited DPP‐IV activity (IC50 1.18 ± 0.04 and 1.21 ± 0.04 mg mL−1), respectively. The SGID sample produced a 1.3‐fold (P < 0.01) increase in GLP‐1 secretion. An oral glucose tolerance test (OGTT) was conducted in healthy mice (n = 8), with or without BPH (50 mg/kg bodyweight). BPH mediated an increase in plasma insulin levels (AUC(0–120 min), P < 0.05) and a consequent reduction in blood glucose concentration (P < 0.01), after OGTT in mice versus controls. The BPH showed potent anti‐diabetic actions in cells and improved glucose tolerance in mice.