Efficient chemo-enzymatic gluten detoxification: reducing toxic epitopes for celiac patients improving functional properties
2015
Ribeiro, Miguel | Nunes, Fernando M. | Guedes, Sofia | Domingues, Pedro | Silva, Amélia M. | Carrillo, Jose Maria | Rodriguez-Quijano, Marta | Branlard, Gerard | Igrejas, Gilberto | CQ-VR, Chemistry Research Centre, Chemistry Department ; University of Tras-os-Montes and Alto Douro | Department of Genetics and Biotechnology ; University of Tras-os-Montes and Alto Douro | Functional Genomics and Proteomics Unity ; University of Tras-os-Montes and Alto Douro | Chemistry Department ; Universidade de Aveiro = University of Aveiro | Centre for the Research and Technology of Agro-Environmental and Biological Sciences, (CITAB-UTAD) ; University of Tras-os-Montes and Alto Douro | Unidad de Genética y Mejora de plantas, Departamento de Biotecnología, E.T.S. Ingenieros Agrónomos ; Universidad Politécnica de Madrid (UPM) | Unidad de Genética y Mejora de plantas, Departamento de Biotecnología, E.T.S. Ingenieros Agrónomos ; Universidad Politécnica de Madrid (UPM) | Génétique Diversité et Ecophysiologie des Céréales (GDEC) ; Institut National de la Recherche Agronomique (INRA)-Université Blaise Pascal - Clermont-Ferrand 2 (UBP) | Functional Genomics and Proteomics Unity ; University of Tras-os-Montes and Alto Douro | Faculty of Science and Technology, BIOSCOPE Group, UCIBIO-REQUIMTE ; Universidade de Lisboa = University of Lisbon = Université de Lisbonne (ULISBOA) | Foundation for Science and Technology (FCT) SFRH/BD/82334/2011, European Social Fund (ESF), FCT PTDC/QUI-QUI/100044/2008 PEst-OE/QUI/UI0616/2014 PEst-C/QUI/UI0062/2013 FCOMP-01-0124-FEDER-037296 REDE/1504/REM/2005, COMPETE
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.
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