Treatment of type 2 diabetes is achieved through the inhibition of carbohydrate hydrolyzing enzymes such as α-glucosidase and α-amylase. The present study was conducted to identify novel α-glucosidase inhibitory peptides and to validate the α-glucosidase and α-amylase inhibitory activities of two promising candidates. A total of 4210 potential α-glucosidase inhibitory peptides with 3–5 amino acid residues were designed and individually subjected to in silico simulated gastrointestinal (GIT) digestion using the BIOPEP database. Subsequently, 844 GIT resistant peptides were then subjected to molecular docking using Autodock Vina to determine their binding free energy against human α-glucosidase (PDB ID: 3L4Y). Among all the peptides, SVPA and SEPA were found to have the lowest binding free energies of -8.7 and -8.6 kcal/mol, respectively. Docking of SVPA and SEPA on human α-amylase (PDB ID, 4GQR) identified that both peptides also bind to α-amylase with binding energies of -6.5 and -6.9 kcal/mol, respectively. Hydrogen bond interactions were critical for the binding of both peptides to the α-glucosidase and α-amylase. In vitro, SVPA and SEPA inhibited α-glucosidase and α-amylase activities with IC₅₀ values several fold lower than acarbose except for SVPA that had a significantly higher (p <  0.05) IC₅₀ value than acarbose against α-glucosidase. Lineweaver-Burk analyses revealed that SVPA was an uncompetitive inhibitor of the two enzymes, while SEPA inhibited α-glucosidase and α-amylase non-competitively and uncompetitively, respectively. This study has identified two novel and active α-glucosidase inhibitory peptides that could resist GIT digestion and therefore, have the potential to retard postprandial hyperglycemia in diabetic patients.