Potential allergenicity and hydrolysis assessment of bovine casein and ß-casein by treatment with lactic acid bacteria.

Casein is one of the main allergens in cow's milk, accounting for 80% of cow's milk proteins. The ability of hydrolyzing proteins by bacteria is also different. In this study, the capacity of lactic acid bacteria to hydrolyze casein or ß-casein and the IgG/IgE-binding capacity of hydrolysates were evaluated. The intensity of casein and ß-casein degradation was analyzed by SDS-PAGE and RP-HPLC. The hydrolysates were tested for their capacity to inhibit IgG and IgE binding by ELISA. The peptides in the hydrolysate were also analyzed by LC-MS/MS. In these strains, Lactobacillus rhamnosus (CICC No. 22175) had the strongest hydrolysis of casein and ß-casein. The hydrolysate of Lactobacillus rhamnosus (CICC No. 22175) showed the lowest antigenicity and potential allergenicity. It also hydrolyzed major allergen IgE epitopes and preserved T cell epitopes. Thereore Lactobacillus rhamnosus (CICC No. 22175) could be used for developing hypoallergenic dairy products and the development of tolerance. PRACTICAL APPLICATIONS: By the study, it obtained that a strain of Lactobacillus rhamnosus could effectively degrade casein and reduced the potential allergenicity of casein. At the same time, some major allergic epitopes were hydrolyzed and T cell epitopes were preserved. Therefore, it is very valuable for the application and development of lactic acid bacteria. The hydrolysate can also be used in a new hypoallergenic dairy formula with specific health benefits and promoting oral tolerance.

Potential allergenicity assessment after bovine apo-α-lactalbumin binding to calcium ion.

Bovine α-lactalbumin (α-LA) is recognized as a major milk allergen. Generally, α-LA in the natural state combines with a calcium ion, however, some studies have shown that calcium ions can binding the other metal binding sites in α-LA as well. In our study, the optimal condition of calcium ion binding to α-LA and the change of structure and allergenicity were explored. By optimizing the conditions, the maximum calcium binding amounts of apo-α-LA were obtained in a ratio of 1:4. The structure of α-LA after removal of calcium obviously changed by the spectroscopic detection. For the digestive stability, there was no obvious change in three forms of α-LA. While the allergenic properties were characterized by IgG/IgE inhibition ELISA and the human basophil KU812 degranulation assay. The results showed that IgG and IgE binding decreased, and the degranulation capacity of basophils weakened. Based on these results, calcium binding to apo-α-LA can reduce the potential allergenic properties. PRACTICAL APPLICATIONS: By optimizing the conditions, bovine apo-α-LA can obtain the most calcium binding amount. And calcium binding to apo-α-LA can reduce the potential allergenic properties. Compared with α-LA in the natural state, calcium binding to α-LA not only can reduce the allergenic properties, but also play a role in calcium supplementation. It might be used to guide the development of hypoallergenic α-LA and provide a method to reduce the potential allergenic properties of α-LA.