The objective was to evaluate the effect of black bean protein fraction (PFRA)， and its derived peptides on glucose uptake， SGLT1 and GLUT2 expression and translocation on Caco-2 cells. The effect of treatments was evaluated on glucose uptake， protein expression and localization and gene expression on Caco-2 cells. PFRA (10 mg/mL) lowered glucose uptake from 27.4% after 30 min to 33.9% after 180 min of treatment compared to untreated control (p < 0.05). All treatments lowered GLUT2 expression after 30 min of treatment compared to untreated control (31.4 to 48.6%， p < 0.05). Similarly， after 24 h of treatment， GLUT2 was decreased in all treatments (23.5% to 48.9%) (p < 0.05). SGLT1 protein expression decreased 18.3% for LSVSVL (100 μM) to 45.1% for PFRA (10 mg/mL) after 24 h. Immunofluorescence microscopy showed a decrease in expression and membrane translocation of GLUT2 and SGLT1 for all treatments compared to untreated control (p < 0.05). Relative gene expression of SLC2A2 (GLUT2) and SLC5A1 (SGLT1) was downregulated significantly up to two-fold change compared to the untreated control after 24 h treatment. Black bean protein fractions are an inexpensive， functional ingredient with significant biological potential to reduce glucose uptake and could be used as an adjuvant in the treatment of colorectal cancer.