Comparative study on the physicochemical and fermentation properties of dough with three fermentation modes under freeze–thaw cycles

BACKGROUND: Frozen fermented dough frequently undergoes temperature fluctuations during distribution, leading to repeated freeze-thaw (FT) cycles that compromise quality. While industrial practices vary between non-fermented and pre-fermented strategies, comparative insights into how pre-fermentation duration dictates dough stability under varying FT stresses remain limited. This study investigated the physicochemical, structural, and rheological evolution of dough subjected to three and six FT cycles across three pre-fermentation modes: non-fermented (0 min), moderate (20 min), and prolonged (50 min). RESULTS: Moderate pre-fermentation (20 min) significantly enhanced microstructural integrity under mild stress (three cycles). This was driven by yeast metabolites (glycerol, glutathione) that reinforced the gluten network, increased β-sheet content, and elevated rheological moduli surpassing even fresh controls. However, six cycles induced progressive deterioration across all groups via ice-driven disruption of hydrogen/hydrophobic bonds, leading to protein unfolding, disulfide cleavage, wet gluten loss, and increased damaged starch. Notably, non-fermented dough (0 min) exhibited superior stability under severe stress, attributed to its compact, gas-free matrix, which mitigated ice expansion and restricted water migration. Conversely, prolonged fermentation (50 min) exacerbated fragility, where compensatory ionic bonding failed to prevent structural collapse. CONCLUSION: FT stability is critically dependent on pre-fermentation time. We propose a dual-phase strategy: moderate pre-fermentation (20 min) is optimal for short-term storage (three cycles) by leveraging metabolite-mediated network reinforcement, whereas minimizing pre-fermentation (0 min) is essential for long-term distribution (six cycles) to preserve structural integrity against ice-induced damage. © 2026 Society of Chemical Industry.