Proteomic Characterization of Wheat Protein Fractions Taken at Different Baking Conditions.

Food processing conditions affect the structure, solubility, and therefore accurate detection of gluten proteins. We investigated the influence of dough, bread, and pretzel making on the composition of different wheat protein fractions obtained by Osborne fractionation. The albumin/globulin, gliadin, and glutenin fractions from flour, dough, crispbread, bread, and pretzel were analyzed using RP-HPLC, SDS-PAGE, and untargeted nanoLC-MS/MS. This approach enabled an in-depth profiling of the fractionated proteomes and related compositional changes to processing conditions (mixing, heat, and alkali treatment). Overall, heat treatment demonstrated the most pronounced effect. Label-free quantitation revealed significant changes in the relative abundances of 82 proteins within the fractions of bread crumb and crust in comparison to flour. Certain gluten proteins showed shifts or reductions in particular fractions, indicating their incorporation into the gluten network through SS and non-SS cross-links. Other gluten proteins were enriched, suggesting their limited involvement in the gluten network formation.

Influence of baking conditions on the extractability and immunochemical detection of wheat gluten proteins.

Food processing conditions affect the accurate detection of gluten by ELISA, which is of importance for proper gluten-free labelling. We prepared different wheat flour-based and incurred baked goods (bread, crispbread, pretzel) to investigate the influence of baking conditions and alkali treatment on gluten quantitation by ELISA using different extraction solvents. Protein composition and extractability were determined (SDS-PAGE, RP-HPLC, GP-HPLC). The extraction solvents showed different performances; none of them could compensate the effect of baking on the detection. Dough preparation, baking and additional alkali treatment decreased protein extractability under reducing and non-reducing conditions. High temperature combined with alkali treatment resulted in the lowest protein extractabilities (<77% for bread crust, <61% for pretzel crust) due to the formation of disulfide and non-disulfide gluten crosslinks. There was no clear correlation between the protein composition and the extractability of alcohol- and SDS-soluble proteins of the baked goods. Thus, this research shows that gluten extractability rather than gluten composition is crucial for detection by ELISA in baked goods.