title: Efficient chemo-enzymatic gluten detoxification: reducing toxic epitopes for celiac patients improving functional properties.
abstract: Protein engineering of gluten, the exogenous effector in celiac disease, seeking its detoxification by selective chemical modification of toxic epitopes is a very attractive strategy and promising technology when compared to pharmacological treatment or genetic engineering of wheat. Here we present a simple and efficient chemo-enzymatic methodology that decreases celiac disease toxic epitopes of gluten proteins improving its technological value through microbial transglutaminase-mediated transamidation of glutamine with n-butylamine under reducing conditions. First, we found that using low concentrations of amine-nucleophile under non-reducing conditions, the decrease in toxic epitopes is mainly due to transglutaminase-mediated cross-linking. Second, using high amine nucleophile concentrations protein cross-linking is substantially reduced. Third, reducing conditions increase 7-fold the transamidation reaction further decreasing toxic epitopes amount. Fourth, using n-butylamine improves gluten hydrophobicity that strengthens the gluten network. These results open the possibility of tailoring gluten for producing hypoallergenic flours while still taking advantage of the unique viscoelastic properties of gluten.
authors.affiliation: CQ-VR, Chemistry Research Centre, Chemistry Department, University of Trás-os-Montes and Alto Douro, 5000-801 Vila Real, Portugal.|Department of Genetics and Biotechnology, University of Trás-os-Montes and Alto Douro, 5000-801 Vila Real, Portugal.|Functional Genomics and Proteomics Unity, University of Trás-os-Montes and Alto Douro, 5000-801 Vila Real, Portugal.
authors.forename: Miguel
authors.initials: M
authors.lastname: Ribeiro
authors.affiliation: CQ-VR, Chemistry Research Centre, Chemistry Department, University of Trás-os-Montes and Alto Douro, 5000-801 Vila Real, Portugal.
authors.forename: Fernando M
authors.initials: FM
authors.lastname: Nunes
authors.affiliation: Chemistry Department, University of Aveiro, 3810-193 Aveiro, Portugal.
authors.forename: Sofia
authors.initials: S
authors.lastname: Guedes
authors.affiliation: Chemistry Department, University of Aveiro, 3810-193 Aveiro, Portugal.
authors.forename: Pedro
authors.initials: P
authors.lastname: Domingues
authors.affiliation: Centre for the Research and Technology of Agro-Environmental and Biological Sciences, University of Trás-os-Montes and Alto Douro (CITAB-UTAD), 5000-801 Vila-Real, Portugal.
authors.forename: Amélia M
authors.initials: AM
authors.lastname: Silva
authors.affiliation: Unidad de Genética y Mejora de plantas Departamento de Biotecnología, E.T.S. Ingenieros Agrónomos Universidad Politécnica de Madrid, 28040 Madrid, España.
authors.forename: Jose Maria
authors.initials: JM
authors.lastname: Carrillo
authors.affiliation: Unidad de Genética y Mejora de plantas Departamento de Biotecnología, E.T.S. Ingenieros Agrónomos Universidad Politécnica de Madrid, 28040 Madrid, España.
authors.forename: Marta
authors.initials: M
authors.lastname: Rodriguez-Quijano
authors.affiliation: Institut National de la Recherche Agronomique GDEC/UBP, UMR 1095, 63100 Clermont-Ferrand, France.
authors.forename: Gérard
authors.initials: G
authors.lastname: Branlard
authors.affiliation: Department of Genetics and Biotechnology, University of Trás-os-Montes and Alto Douro, 5000-801 Vila Real, Portugal.|Functional Genomics and Proteomics Unity, University of Trás-os-Montes and Alto Douro, 5000-801 Vila Real, Portugal.|BIOSCOPE Group, UCIBIO-REQUIMTE, Chemistry Department, Faculty of Science and Technology, University NOVA of Lisbon, Caparica, Portugal.
authors.forename: Gilberto
authors.initials: G
authors.lastname: Igrejas
mesh_terms: D000596:Amino Acids / Q000032:analysis; D018938:Caco-2 Cells; D002446:Celiac Disease / Q000473:pathology*; D016923:Cell Death; D003432:Cross-Linking Reagents / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D000939:Epitopes / Q000009:adverse effects*; D005433:Flour; D008401:Gas Chromatography-Mass Spectrometry; D005983:Glutens / Q000009:adverse effects* / Q000737:chemistry*; D006801:Humans; D010084:Oxidation-Reduction; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism; D040901:Proteomics; D012212:Rheology; D053719:Tandem Mass Spectrometry; D014908:Triticum / Q000009:adverse effects
chemical_list: D000596:Amino Acids; D003432:Cross-Linking Reagents; D000939:Epitopes; D021122:Protein Subunits; D005983:Glutens