Unveiling the Mechanism of High-Molecular-weight Glutenin Subunit Deletions at the Glu-B1 Locus Affecting Gluten Deterioration During Dough Frozen Storage and Freeze-Thaw Cycles: an Integrative Experimental and in Silico Study

This study explored how high-molecular-weight glutenin subunits (HMW-GS) at the Glu-B1 locus impact gluten deterioration during dough frozen storage and freeze-thaw (F/T) cycles. Using deletion lines, we found that the deletion of specific HMW-GS, particularly Bx7, led to a greater reduction in glutenin macropolymer (GMP) wet weight during storage, especially under F/T cycles. The frozen conditions triggered more dissociation of hydrogen and disulfide bonds, generating more protein monomers and resulting in severe gluten deterioration in the Bx7 deletion. Additionally, protein structures in these lines were more vulnerable to damage during F/T cycles due to temperature fluctuations. In silico analysis further confirmed that Bx7 had better stability and antifreeze activity compared to By8, explaining why its deletion had a more pronounced effect on gluten stability. These findings offer significant implications for the food industry, particularly in enhancing the quality, shelf-life, and commercial viability of frozen dough products by providing a deeper understanding of the mechanisms behind gluten deterioration, specifically the role of high-molecular-weight glutenin subunits.