Proteolytic enzymes were screened to obtain anchovy protein hydrolysates (APH) with low-bitterness under pressurization. First, the residual proteolytic activity (%) and SDS-PAGE of commercially available food-grade protease (Alcalase® 2.4 L, FlavourzymeTM 500 MG, Neutrase®, ProtamexTM) and mixed enzyme (ME, Alcalase® : FlavourzymeTM = 1 : 5) after treatments of ambient pressure and 100 MPa at 50oC were investigated. The proteolytic activity of enzymes was still largely retained after pressure treatment. The SDS-PAGE on tested enzymes showed that the electrophoretic mobility maps (EMMs) after pressure treatment were similar to those at ambient pressure. With these results, it seemed that tested enzymes were stable at 100 MPa and 50oC. Raw anchovy hydrolyzed with various protease at 50oC under ambient pressure and 100 MPa for 1, 2, 6, 12, and 24 h. The degree of hydrolysis (DH, %) of prepared APHs at 100 MPa was increased, which suggests that the medium pressure treatment accelerates protein hydrolysis. The APH by ME exhibited the best suitable characteristics, which had a relatively higher DH (36.2±1.8- 57.5±2.8), a high content of oligopeptides with a molecular mass lower than 1,100 Da, mildly bitter taste as the bitterness value was lower than that of 0.2 g/L tea alkaloid, and the lowest contents of tryptophan. Thus, ME was selected to obtain APH with low-bitterness under pressurization.