Diabetes mellitus is a multifactorial chronic disease that affects the human population and it is the third most common cause of death worldwide. Momordica charantia is commonly known as Bitter melon, Bitter guard and used as a food and natural medicine. The scientific name, Momordica means "to bite," in Latin which is refers to the jagged edges of the leaves. Including fruits, all parts of the plant, contains a bitter compound, momordicinso and very bitter in taste. It has long been used as a traditional medicine for some ailments. Its high protein content (35.18%) makes it a promising candidate as a source of bioactive protein hydrolysates. In this study, the potential of enzymatically hydrolyzed Bitter guard seed protein to generate functional bioactive peptides was evaluated. Bitter guard seed protein concentrate was hydrolyzed using three different proteolytic enzymes (Alcalase, Bromelain, and Papain) at their respective optimum pH and temperature. The choice of enzyme affected the bioactivities to a certain degree as Bitter guard seed were shown to possess high antidiabetic activity. Bromelain Bitter guard seed protein hydrolysate showed the highest DPP-IV inhibitory activity with 79.27% inhibition, α-Glucosidase inhibitory activity with 72.15% inhibition and α-Amylase inhibitory activity with 59.98% compared to other enzymatic hydrolysates, and therefore was chosen for further investigation of its antidiabetic activity.