Aiming to boost the high-value utilization of colored barley resources and to broaden its processing depth in cereal products. This study compared the techno-functional characteristics of hordein/glutelin, a key component for gluten in white (W), blue (B), and black (BK) barley, and attempted to modulate their ratios for enhancing reconstituted dough thermal processing suitability. Results indicated that white barley's hordein/glutelin techno-functional profiles are more adapted for dough processing. During dough thermal processing, C2-C3 interval changes were mainly reflected in morphological structure, water distribution, rheological behavior, and textural properties, while the C3-C4 interval variations were interpreted as protein secondary/tertiary structural transitions and intermolecular interactions. Notably, “hordein: glutelin = 50:50″ was a crucial factor in determining the barley dough quality, i.e., low hordein ratio (25:75) was unfavorable for gluten matrix and continuous network formation, whereas its excessive ratio (75:25) weakened the protein aggregation behavior and gluten interaction with starch. Correlation analysis showed that the gluten network's connection density/porosity, protein's secondary structure, and ionic bond strength are key factors modulating dough processing suitability. Therefore, by rationally regulating the hordein/glutelin ratio, barley dough products can be produced with various edible characteristics for consumption. Also, attention should be focused on breeding high hordein ratio barley kernels to improve their gluten profile and thus promote its potential application in cereal processing.
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