The current work mainly focused on the determination of the bromelain effect on glucose uptake through the everted gut sac technique. Everted gut sacs are majorly used in evaluating the uptake mechanisms and absorption potential. Bromelain is a natural phytochemical obtained from pineapple. It can alter various body mechanisms. Everted gut sacs were prepared from chick ileum. After thorough cleaning, the gut sacs were filled with Krebs-Henseleit bicarbonate buffer (KHB). Glucose at varying concentrations was added into mucosal compartment fluid. Bromelain was also added to the mucosal compartment with simulated gastric fluid. At time intervals of 5, 10, 15, 20, 30, 45, and 60 min, 0.5ml of samples were collected from the mucosal fluid for glucose level estimation. A kinetic study was also conducted to understand the transport/inhibition of glucose across the intestinal membrane. Incubation of the chick-everted gut sacs with Bromelain at concentrations of 10 and 20μg/mL increased the transport of glucose. The glucose transport was significant at various concentrations i.e., 5.5, 6.5, 7.5, and 8.5 mM/L in the incubation medium. The pattern of glucose uptake ex-vivo was analyzed using Michaelis-Menten and Lineweaver-Burk Plots. When compared to control, Km has decreased, but Vmax remained unchanged in the presence of Bromelain at a dose of 10μg/ml. Whereas in the case of Bromelain given at a dose of 20μg/mL, the Vmax has increased and Km remained constant when compared with control. Thus, the current research indicated that Bromelain at various doses increased glucose uptake.