Peanut digestome: Identification of digestion resistant IgE binding peptides

• Published in: Food and chemical toxicology Vol. 107; no. Pt A; pp. 88 - 98
• Main Authors: Di Stasio, Luigia, Picariello, Gianluca, Mongiello, Mariantonietta, Nocerino, Rita, Berni Canani, Roberto, Bavaro, Simona, Monaci, Linda, Ferranti, Pasquale, Mamone, Gianfranco
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.09.2017
• Subjects: allergenicity; allergens; Allergy; Amino Acid Sequence; Antigens, Plant - chemistry; Antigens, Plant - genetics; Antigens, Plant - immunology; Antigens, Plant - metabolism; Arachis - chemistry; Arachis - genetics; Arachis - immunology; Arachis - metabolism; Brush border membrane enzymes; digestibility; Digestion; digestive tract; Electrophoresis, Polyacrylamide Gel; enzymes; food matrix; foods; gastrointestinal system; Humans; hydrolysis; IgE binding peptide; immune response; immunoglobulin E; Immunoglobulin E - immunology; In vitro human digestion; Mass spectrometry-based proteomics; microvilli; Molecular Sequence Data; Peanut; Peanut Hypersensitivity - immunology; Peanut Hypersensitivity - metabolism; peanuts; Peptide Mapping; peptides; Peptides - chemistry; Peptides - genetics; Peptides - immunology; polyacrylamide gel electrophoresis; proteolysis; reversed-phase high performance liquid chromatography; Tandem Mass Spectrometry; toxicology; Western blotting; Allergy; Peanut; TAME; DTT; SSF; IAA; BBM; Brush border membrane enzymes; TFA; IgE binding peptide; SIF; TCA; SGF; Ambic; In vitro human digestion; Mass spectrometry-based proteomics
• ISSN: 0278-6915; 1873-6351; 1873-6351
• DOI: 10.1016/j.fct.2017.06.029

Stability to proteolytic degradation in the digestive tract is considered a general feature shared by most food allergens. Current digestibility models exclusively utilize purified allergen proteins, neglecting the relevant effects of matrix that occur for foodstuff systems. In the present study, we investigated digestion stability of the major peanut allergens directly in the natural matrix using an in vitro static model that simulates the gastrointestinal digestion including the oral, gastric, duodenal and intestinal (brush border membrane enzymes) phases. Immunogenicity was evaluated by Western Blot using N=8 pooled sera of peanut allergic pediatric subjects. Immunoreactive, large-sized and fragments of Ara h 2, Ara h 6 and Ara h 3 survived hydrolysis as assessed by SDS-PAGE. Smaller resistant peptides mainly arising from Ara h 3 and also Ara h 1 were detected and further identified by LC-high resolution-MS/MS. RP-HPLC purification followed by dot-blot analysis and MS/MS-based identification demonstrated that stable IgE-binding peptides derived from Ara h 3. These results provide a more realistic picture of the potentially allergenic determinants of peanuts that survived the human digestion, taking into account the role of the food matrix, which may significantly affect gastrointestinal breakdown of peanut allergens. [Display omitted] •A realistic picture of the potentially allergenic peanuts is provided.•Large fragments of Ara h 3, survive to in vitro gastrointestinal digestion.•Ara h 3 IgE binding epitopes are released during gastrointestinal digestion.•Ara h 1 is completely degraded following in vitro gastrointestinal digestion.•Peanut matrix slows the gastrointestinal digestion process of allergen proteins.