Cross-linking of shrimp tropomyosin catalyzed by transglutaminase and tyrosinase produces hypoallergens for potential immunotherapy.

Tropomyosin (TM) is the major allergen in shellfish, which causes severe food allergy worldwide and shows wide cross-reactivity in different species. Transglutaminase (TG) and tyrosinase (Tyr) are protein modification enzymes that catalyze substrate cross-linking and change protein structures and properties. The aim of this work was to investigate the effect of TG and Tyr on TM. After TG/Tyr treatment, the cross-linking and protein structural change were observed by SDS-PAGE, scanning electron microscopy and circular dichroism. Furthermore, ELISA and mast cell degranulation assay with allergic sera demonstrated the allergenicity reduction after treatment. An in vivo TM-sensitization mouse model showed that both TG and Tyr treatments reduced the IgE-induction capacity of TM but maintained the IgG-induction ability, making the treated TM a potential hypoallergen. Moreover, TG-but-not-Tyr-treated TM promoted regulatory T cell proliferation and thus may facilitate the establishment of oral tolerance. This work provided an effective method to reduce TM allergenicity and produce hypoallergens, which has broad application prospects in low allergenic food production and allergen-specific immunotherapy.

Maillard reaction with ribose, galacto-oligosaccharide or chitosan-oligosaccharide reduced the allergenicity of shrimp tropomyosin by inducing conformational changes.

Tropomyosin (TM), the predominant shellfish allergen, has induced severe food allergy worldwide, but efficient prevention and cure measures are limited nowadays. In the present study, we introduced an approach to eliminate Penaeus chinensis TM by Maillard reaction with reducing sugars including ribose, galacto-oligosaccharide (GOS) and chitosan-oligosaccharide (COS), which reduced the allergenicity by up to 60%. We also evaluated the conformational changes of TM during reaction by far-UV circular dichroism, and found the reaction-induced alternation from α-helix to ß-sheet. We further determined the effect of Maillard reaction on TM allergenic epitopes to investigate the detailed mechanisms, and revealed the structure preference of different sugars. Correlation analysis with experimental data revealed that α-helix mediated the allergenicity elimination by Maillard reaction. There results provided theoretical foundations for the improvement and application of Maillard reaction in allergenic food processing.

Identification of Allergenic Epitopes and Critical Amino Acids of Major Allergens in Chinese Shrimp ( Penaeus chinensis) by Immunoinformatics Coupled with Competitive-Binding Strategy.

Chinese shrimp ( Penaeus chinensis) is widely cultured and consumed in Asia but is also a major food allergen locally. Although they may be the foundation for preventing and treating allergies, the allergenic epitopes of the major allergens tropomyosin (TM) and arginine kinase (AK) in Penaeus chinensis have not been identified. Here, we applied Immunoinfo-CB (immunoinformatics coupled with competitive-binding strategy) to address the point. Potential allergenic epitopes of TM and AK were predicted by multiple immunoinformatics tools, followed by validating with inhibitory dot-blot assay, indirect competition ELISA, and mast cell degranulation assay. Furthermore, critical amino acids in allergenic epitopes were also identified by Immunoinfo-CB. Our findings provide new insight into allergenic epitopes and critical amino acids of TM and AK responsible for the anaphylactic response. The Immunoinfo-CB therefore offers promises for characterization of IgE-binding epitopes that might be used as new targets for immunotherapy of food allergy.