Influence of bile salts on the gastrointestinal digestion of agar-casein hybrid systems and the nanoassembly of their digestion products

• Agar-casein hydrogels reduce the proteolysis upon gastrointestinal digestion. • Bile salts promote casein hydrolysis. • Bile salts promote a greater diffusion of casein from the hydrogels. • Agar and casein peptides interfere with the bile salts nanoassembly into lamellae. • The presence of soluble agar in the digesta stabilizes bile salt lamellae. This manuscript reports on the effect of different bile salts concentration on the gastrointestinal digestion of casein and casein-agar hybrid systems and evaluates the effect on the nanostructural assembly of the digestion products through the application of advanced small angle X-ray scattering analysis (SAXS). The results showed that bile salts promote the hydrolysis of micellar casein upon in vitro gastrointestinal digestions. It is noteworthy that in the presence of agar, bile salts interact with the polysaccharide, leading to a reduction in their proteolytic activity. While structured agar-casein hydrogels were shown to be able to limit the proteolysis during the gastric phase, increasing concentrations of bile salts promoted the diffusion of casein from the gel network during the intestinal phase, hence leading to higher degree of hydrolysis. The released digestion products were seen to interact with bile salts, forming ordered lamellar/micellar nanostructures. In particular, the presence of solubilized agar in the digesta was seen to promote the formation of these nanostructures. These nanostructures were formed at 10 mM bile salts for the hydrogels, and across all concentrations for agar-casein blends. Thus, this work demonstrates the great relevance of bile salts in the nanostructural assembly of protein digestion products, which is expected to have a great relevance in bioavailability and metabolic responses induced by protein-rich foods.