Extensive hydrolysis of milk protein and its peptidomic antigenicity analysis by LC–MS/MS

Abstract

Milk protein concentrate (MPC) is considered an ideal substitute of cow milk because of its similar protein and nutrition. In this study, MPC was hydrolyzed to peptides by alcalase and neutrase, and the properties of hydrolysate were evaluated. When MPC was hydrolyzed at the ratio of alcalase and neutrase of 1:1 and enzyme to substrate ratio of 6000 U/g MPC at 50°C, pH 8.5 for 3 h, the proportion of peptides with molecular weights <1 kDa was 85.31%, and the antigenicity reduction rates of casein and β‐lactoglobulin were 33.76% and 22.38%, respectively. Moreover, LC–MS/MS peptide identification revealed that the alcalase and neutrase disrupted a total of 65 epitopes of casein and 21 epitopes of whey protein, which further elucidated the mechanism of complex enzyme hydrolysis to reduce milk protein allergenicity.

One-Step Purification of IgE Epitope-Specific Antibody Using Immunomagnetic Beads and Highly Sensitive Detection of Bovine β-Lactoglobulin for the Prediction of Milk Allergenicity in Foods

“…Furthermore, the low LOD (0.49 ng/mL) and ultra-wide linear working range (3.9–1.28 × 10 5 ng/mL) of our method made it possible to mitigate the matrix effects of real food samples with appropriate dilution, without compromising the sensitivity. , Significantly, the IgE-EsAb used in our sELISA could specifically recognize the major IgE epitope of BLG, enabling the specific detection of BLG allergenic residues in processed foods . Although LC–MS/MS can also detect BLG epitopes, it lacks the sensitivity and broad linear working range of our sELISA . Additionally, LC–MS/MS requires specialized knowledge, expensive equipment, and a lengthy sample pretreatment process …”

“…12 Although LC−MS/ MS can also detect BLG epitopes, it lacks the sensitivity and broad linear working range of our sELISA. 39 Additionally, LC− MS/MS requires specialized knowledge, expensive equipment, and a lengthy sample pretreatment process. 40 Lastly, the accuracy and precision of the IgE-EsAb-based sELISA were evaluated.…”

Characterization of the potential allergenicity of enzymatically hydrolyzed casein in Balb/c mouse model

“…Similarly, some research showed that Alcalase and Protamex were effective in reducing the antigenicity of CN and β-LG and significantly superior to other enzymes (Liang et al, 2021). Li et al (2023) studied the antigenicity of milk protein concentrate treated with Alcalase and Neutrase and found that the antigenicity reduction of CN and β-LG reached 33.76% and 22.38% (Li et al, 2023). Additionally, for the FT group, the antigenicity inhibition rate of CN reduced rapidly during the first 60 min, reaching the lowest (from 0.88% to −15.80%).…”