Hypertension, a non-communicable disease affecting over one third of the world population, is a risk factor for heart diseases. Thus developing functional foods with ability of alleviating hypertension is of great importance to maintain body health in a natural way.Sea cucumbers,belonging to the class of Holothuroidea, are rich sources of nutrients and bioactive compounds and there is a strong belief upon their various health benefits. Angiotensin-I converting enzyme (ACE) is a key enzyme of the renin-angiotensin-aldosterone system,and responsible for the elevation of the blood pressure. Therefore, production of natural, safe and effective ACE inhibitors is a tall order. Due to the advantages of food-derived bioactive peptides, this work was aimed at generating bioactive peptides through enzymatic proteolysis of S. horrens flesh using different proteases. The proteolysate was further purified and characterized for ACE-inhibitory activity. Among the six (6) different proteases employed,alcalase exhibited the highest ability to proteolyse and generate ACE-inhibitory peptides followed by flavourzyme with IC50 values of 0.41 and 2.24 mg/mL, respectively. Fractionation of the effective peptides was carried out based on their hydrophobicities and isoelectric points.The most effective fraction was subjected to UPLC-MS/MS for peptides identification. A total of 24 peptides were identified showing 4 to 16 amino acid residues (molecular weights between453.4 and 1771.9 kDa) withIC50values in the range of 0.05-8.8 mM. The Kinetics studyof the peptides revealed amixed-mode inhibition pattern.This study has demonstratedS. Horrens as a suitable marine source for ACE-inhibitory peptides generation.