title: Maillard reaction and enzymatic browning affect the allergenicity of Pru av 1, the major allergen from cherry (Prunus avium).
abstract: The influence of thermal processing and nonenymatic as well as polyphenoloxidase-catalyzed browning reaction on the allergenicity of the major cherry allergen Pru av 1 was investigated. After thermal treatment of the recombinant protein rPru av 1 in the absence or presence of carbohydrates, SDS-PAGE, enzyme allergosorbent tests, and inhibition assays revealed that thermal treatment of rPru av 1 alone did not show any influence on the IgE-binding activity of the protein at least for 30 min, thus correlating well with the refolding of the allergen in buffer solution as demonstrated by CD spectroscopic experiments. Incubation of the protein with starch and maltose also showed no effect on IgE-binding activity, whereas reaction with glucose and ribose and, even more pronounced, with the carbohydrate breakdown products glyceraldehyde and glyoxal induced a strong decrease of the IgE-binding capacity of rPru av 1. In the second part of the study, the effect of polyphenoloxidase-catalyzed oxidation of polyphenols on food allergen activity was investigated. Incubation of rPru av 1 with epicatechin in the presence of tyrosinase led to a drastic decrease in IgE-binding activity of the protein. Variations of the phenolic compound revealed caffeic acid and epicatechin as the most active inhibitors of the IgE-binding activity of rPru av 1, followed by catechin and gallic acid, and, finally, by quercetin and rutin, showing significantly lower activity. On the basis of these data, reactive intermediates formed during thermal carbohydrate degradation as well as during enzymatic polyphenol oxidation are suggested as the active chemical species responsible for modifying nucleophilic amino acid side chains of proteins, thus inducing an irreversible change in the tertiary structure of the protein and resulting in a loss of conformational epitopes of the allergen.
authors.affiliation: Deutsche Forschungsanstalt für Lebensmittelchemie, Lichtenbergstrasse 4, D-85748 Garching, Germany.
authors.forename: Patrick
authors.initials: P
authors.lastname: Gruber
authors.forename: Stefan
authors.initials: S
authors.lastname: Vieths
authors.forename: Andrea
authors.initials: A
authors.lastname: Wangorsch
authors.forename: Jörg
authors.initials: J
authors.lastname: Nerkamp
authors.forename: Thomas
authors.initials: T
authors.lastname: Hofmann
mesh_terms: D000485:Allergens / Q000737:chemistry* / Q000276:immunology*; D052179:Antigens, Plant; D005511:Food Handling; D006358:Hot Temperature; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood / Q000276:immunology; D015416:Maillard Reaction; D027861:Prunus / Q000276:immunology*
chemical_list: D000485:Allergens; D052179:Antigens, Plant; C479400:Pru av 1 allergen, Prunus avium; D007073:Immunoglobulin E