The Fate of IgE Epitopes and Coeliac Toxic Motifs during Simulated Gastrointestinal Digestion of Pizza Base
Matthew E. Daly,
Kai Wang,
Xiaoyan Pan,
Rosa L. Depau,
Justin Marsh,
Francesco Capozzi,
Phil Johnson,
Lee A. Gethings,
E. N. Clare Mills
Affiliations
Matthew E. Daly
Manchester Institute of Biotechnology, School of Biological Sciences, Manchester Academic Health Sciences Centre, The University of Manchester, Princess Street, Manchester M1 7DN, UK
Kai Wang
Manchester Institute of Biotechnology, School of Biological Sciences, Manchester Academic Health Sciences Centre, The University of Manchester, Princess Street, Manchester M1 7DN, UK
Xiaoyan Pan
Manchester Institute of Biotechnology, School of Biological Sciences, Manchester Academic Health Sciences Centre, The University of Manchester, Princess Street, Manchester M1 7DN, UK
Rosa L. Depau
Manchester Institute of Biotechnology, School of Biological Sciences, Manchester Academic Health Sciences Centre, The University of Manchester, Princess Street, Manchester M1 7DN, UK
Justin Marsh
Manchester Institute of Biotechnology, School of Biological Sciences, Manchester Academic Health Sciences Centre, The University of Manchester, Princess Street, Manchester M1 7DN, UK
Francesco Capozzi
Department of Food Sciences, University of Bologna, 40126 Bologna, Italy
Phil Johnson
Manchester Institute of Biotechnology, School of Biological Sciences, Manchester Academic Health Sciences Centre, The University of Manchester, Princess Street, Manchester M1 7DN, UK
Lee A. Gethings
Manchester Institute of Biotechnology, School of Biological Sciences, Manchester Academic Health Sciences Centre, The University of Manchester, Princess Street, Manchester M1 7DN, UK
E. N. Clare Mills
Manchester Institute of Biotechnology, School of Biological Sciences, Manchester Academic Health Sciences Centre, The University of Manchester, Princess Street, Manchester M1 7DN, UK
Understanding how food processing may modify allergen bioaccessibility and the evolution of immunologically active peptides in the gastrointestinal tract is essential if knowledge-based approaches to reducing the allergenicity of food are to be realised. A soy-enriched wheat-based pizza base was subjected to in vitro oral–gastro–duodenal digestion and resulting digests analysed using a combination of sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometry (MS). The digestion profile of pizza base resembled that of bread crust where higher temperatures during baking reduced protein solubility but still resulted in the generation of a complex mixture of peptides. MS profiling showed numerous peptides carrying IgE epitopes, and coeliac toxic motifs were in excess of 20–30 residues long and were only released after either 120 min of gastric digestion or a combination of gastric and duodenal digestion. In silico prediction tools showed an overestimated number of cleavage sites identified experimentally, with low levels of atypical peptic and chymotryptic cleavage sites identified particularly at glutamine residues. These data suggest that such alternative pepsin cleavage sites may play a role in digestion of glutamine-rich cereal foods. They also contribute to efforts to provide benchmarks for mapping in vitro digestion products of novel proteins which form part of the allergenicity risk assessment.