Fluorescence labeling of wheat proteins for determination of gluten hydrolysis and depolymerization during dough processing and sourdough fermentation

This study was undertaken to enable the determination of hydrolysis and functionality of proteins in situ during fermentation of wheat doughs. Wheat proteins were fractionated and labeled with fluorescein-isothiocyanate (FITC). Fluorescent proteins were incorporated into wheat sourdoughs inoculated with lactobacilli and into neutral and acid control doughs. Doughs containing fungal protease were furthermore evaluated. Doughs were analyzed by extraction and size exclusion chromatography analysis of sodium dodecyl sulfate soluble proteins. Labeled proteins exhibited characteristics comparable to native proteins, with respect to proteolytic degradation and polymerization. Proteolytic breakdown of proteins was enhanced at low pH. Glutenin subunits were incorporated into the gluten macropolymer at neutral pH. Polymerization of FITC proteins was not observed at low pH. Sourdoughs were comparable to acid control doughs, major effects were attributed to changes of pH, rather than microbial metabolism. A synergistic effect with respect to proteolytic activity was observed between fungal protease and L. pontis.