Egg White Ovalbumin Digestion Mimicking Physiological Conditions

Gastrointestinal digestion of ovalbumin (OVA) was simulated using an in vitro system in two steps, which mimicked digestion in the stomach and duodenum, to assess the effect of different gastric pHs, different concentrations of proteases, and the presence of surfactants, such as phosphatidylcholine (PC) and bile salts (BS). OVA was very resistant to pepsin action at an enzyme/substrate ratio that would resemble a physiological situation (1:20 w/w, 172 units/mg) at pH values equal or above 2. The presence of PC did not change the susceptibility of OVA to proteolysis with pepsin. Fluorescence experiments showed that OVA interacted with PC vesicles, particularly at acidic pH, but it is likely that the protein maintained a high degree of conformational stability, resisting pepsin action. The presence of BS at physiological concentrations considerably increased the proteolysis of OVA by a mixture of pancreatic enzymes. The addition of PC made OVA even more sensitive to proteolytic degradation, suggesting that OVA could associate with the surfactants under duodenal conditions, increasing its exposure to pancreatic proteinases. Immunoreactivity against IgE from sera of allergic patients was retained after in vitro gastric digestion, depending on the reactivity of the sera, but it decreased considerably after in vitro duodenal digestion.