Barley is gaining popularity worldwide due to its health-promoting properties and germination can effectively enhance its function through enrichment of some phytochemicals, such as phenolic compounds. To better explore the potential of barley seedling as a functional food ingredient, in the present study, the profile and in vitro antioxidant properties of phenolic compound extract from barley germinated under NaCl stress were investigated. A total of 37 kinds of phenolic compounds were identified in phenolic compound extract by a liquid chromatography-tandem mass spectrometry system. Meanwhile, phenolic compound extracts at a concentration of 30 mu g/mL significantly improved the viability of H2O2-treated HepG2 cells and maintained the cell membrane stability. These effects can be attributed to the reduction of intracellular reactive oxygen species level and enhancement of the expression and activity of some key antioxidant enzymes. Notably, addition of phenolic compound extracts up-regulated the nuclear factor erythroid 2-related factor 2 antioxidant responsive element signaling pathway, thereby leading to the increase of activities of glutathione peroxidase, superoxide dismutase, catalase and heme oxygenase-1 by 27.21%, 24.61%, 61.71% and 50.43%, respectively. In conclusion, the present study can provide a novel perspective on understanding the molecular mechanism responsible for antioxidant capacity of barley seedling. (c) 2026 The authors. Publishing services by Elsevier B.V. on behalf of KeAi Communications Co., Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

To balance nutrition and technofunctionality in wheat bread, the wheat aleurone layer was fermented with an exopolysaccharide-producing lactic acid bacterium. Results showed that sourdough without and with in situ exopolysaccharides significantly increased water-extractable arabinoxylans in the wheat aleurone layer by 42.31% and 40.38%, respectively. Enzymatic hydrolysis and exopolysaccharide reverse-addition experiments confirmed that in situ exopolysaccharides primarily contributed to bread quality enhancement, yielding the largest loaf volume by 3.57 mL/g. Wheat aleurone layer sourdough without exopolysaccharides enhanced gluten development and extensibility, whereas sourdough with in situ exopolysaccharides exhibited a 20.75% higher ratio of tensile strength to elongation than sourdough without exopolysaccharides. Acidification and proteolysis depolymerized glutenin macropolymers by 37.79%; however, in situ exopolysaccharides reduced depolymerization to 25.50% and maintained the highest glutenin macropolymer content of 11.63%. Overall, exopolysaccharides effectively stabilized gluten structure and suppressed glutenin macropolymer depolymerization during both mixing and fermentation, thereby enhancing the technofunctionality of high-fiber cereal products.

The succulent xerophyte Pugionium cornutum efficiently translocates chloride ions (Cl-) absorbed by the roots to its shoots for osmotic adjustment under salt stress, a trait uncommon in most crops. However, the underlying molecular mechanisms remain further investigated. The slow-type anion channel AtSLAH3 is known to be involved in Cl- transport from roots to shoots through its interaction with AtSLAH1 in Arabidopsis, but only under non-saline conditions. Here, we investigated the function of its homolog in P. cornutum, PcSLAH3, under saline conditions. The results showed that PcSLAH3, which encodes a plasma membrane-localized protein, was expressed predominantly in the stelar tissues of roots, and exhibited significantly upregulated transcript levels in roots under NaCl treatments. Interactions were observed between PcSLAH3 and PcSLAH1, PcSLAH3, and AtSLAH1, as well as AtSLAH3 and PcSLAH1, suggesting the conservation of the SLAH1-SLAH3 interaction between salt-sensitive and salt-tolerant species. Heterologous expression of PcSLAH3 driven by a root stelar-specific promoter not only significantly increased the Cl- concentration, but also increased the Na+ concentration in shoots of wild-type Arabidopsis or atslah3 mutant under salt treatment. This was accompanied by the upregulated expression of AtCLCg and AtNHX1, which mediate the vacuolar compartmentalization of these two ions. Notably, PcSLAH3 outperformed AtSLAH3 in promoting root-to-shoot Cl- transport, potentially explaining the differences in Cl- transport capacity between xerophytes and glycophytes. These findings demonstrate that PcSLAH3 participates in Cl- transport from roots to shoots and is involved in regulating Cl-/Na+ homeostasis in shoots under saline conditions.

To elucidate the mechanism by which carboxymethyl chitosan (CMCh) regulates heat-induced aggregation of gluten proteins, CMCh derivatives with distinct substitution sites (amino, hydroxyl, and dual amino/hydroxyl substitution) were prepared and systematically compared. The results demonstrated that CMCh effectively modulated gluten aggregation during heating, and the regulatory effect strongly dependent on the substitution site. The degree of substitution (DS) of the derivatives ranged from 0.58 for O-CMCh to 1.26 for N-CMCh, while N,O-CMCh exhibited a DS of 1.18 and the highest water solubility (67.98 g L-1). The dual substituted CMCh (N, O-CMCh) exhibited the most pronounced effect, significantly enhancing gluten network elasticity and thermal stability. The onset and peak denaturation temperatures increased to 62.56 degrees C and 67.88 degrees C, respectively, with a denaturation enthalpy of 0.70 J/g at 95 degrees C, markedly higher than that of the control. Furthermore, N,O-CMCh promoted more ordered disulfide cross-linking during heating, increasing the SDS-insoluble protein content (SDS-I) to 80.40% at 95 degrees C. This was accompanied by a controlled decrease in free sulfhydryl groups and a corresponding increase in disulfide bond formation. In contrast, the single-site substituted derivatives exhibited weaker regulatory effects: N-CMCh increased SDS-I to 75.20%, whereas O-CMCh displayed thermal behavior most similar to the control. The superior functionality of N,O-CMCh is attributed to its unique molecular structure, which enables synergistic electrostatic interactions, hydrogen bonding, and synergistic disulfide crosslinking.This study clarifies the substitution site-dependent structure-function relationship of CMCh and provides a theoretical foundation for the rational design of high-performance CMCh improvers for cereal food.

Excessive dietary sodium is a global health concern, prompting growing interest in ion substitution strategies for sodium reduction without compromising food quality. This study advances the field by systematically deconvoluting how a spectrum of cations (Na+, K+, Ca2+, Mg2+) differentially target the two starch components-amylose and amylopectin-across both gelatinization and retrogradation, through an integrated multiscale approach. Here, we elucidate how four cations-Na+, K+, Ca2+, and Mg2+-govern the gelatinization and retrogradation of wheat starch through distinct modulation of amylose and amylopectin. Multiscale experiments coupled with all-atom molecular dynamics (MD) simulations reveal ion-specific pathways linking hydration thermodynamics to chain dynamics. While all cations modified starch properties, K+ and Mg2+ exerted the most pronounced effects, enhancing solubility while suppressing swelling, viscosity, and recrystallization. In contrast, Ca2+ uniquely moderated gel strengthening without severely inhibiting pasting, whereas Na+ served as a baseline with minimal structural perturbation. Thermal and structural analyses showed higher gelatinization temperatures and 26-39 % lower retrogradation enthalpy and crystallinity, indicating stabilized amorphous regions. MD simulations uncovered the molecular origin of these effects: Mg2+ induces compact, low-entropy conformations in both amylose and amylopectin via strong coordination and dehydration. In strong contrast, K+ primarily maintains flexible, hydrated helices in amylopectin through dynamic hydrogen-bond exchange. Notably, Na+ acted as a weak, non-specific perturbator, while Ca2+ preferentially coordinated with amylopectin, promoting local ordering without inducing the global compaction observed with Mg2+. These ion-dependent conformational landscapes reconcile experimental observations and reveal that valence and hydration energy cooperatively reshape the starch hydrogen-bond network and free-energy surface. This study establishes a unified molecular mechanism linking atomic-scale ion coordination to macroscopic starch stabilization, providing theoretical guidance for the rational design of sodium-reduced starch-based foods with tunable gelatinization and storage behavior.

Self-assembled proteins can significantly inhibit ice recrystallization, offering potential for cryoprotection. Here, soybean protein amyloid fibrils (SAFs) were fabricated via combined germination and acid-heat-induced fibrillation. Germination enhanced the fibrillation efficiency of soybean protein isolate (SPI). SAFs with the strongest ice recrystallization inhibition (IRI) activity were prepared from SPI of two-day germinated soybeans after 20 h of acidic-heat treatment (SAF-20). SAF-20 exhibited concentration-dependent IRI activity, with stronger inhibition of ice crystal growth at higher concentrations. It showed high ice-affinity adsorption and ice nucleation activity without altering ice crystal morphology. Structural analyses revealed that self-assembly promoted protein aggregation and increased surface hydrophobicity and β-sheet content. These changes strengthened hydrogen bonding at the ice-water interface, forming ordered interfacial water layers that disrupted long-range water ordering and inhibited ice crystal growth. Furthermore, SAF-20 significantly improved post-thaw recovery of cryopreserved Caco-2 cells, demonstrating its cryoprotective efficacy.

Wheat seedlings are rich in γ-aminobutyric acid (GABA), phenolic compounds, flavonoids, folate, and chlorophyll. This review covers their targeted enrichment via strategies such as hypoxia stress, red light treatment, and elicitor application, which activate key metabolic pathways. The associated health benefits include sleep improvement, antioxidant and anti-inflammatory effects, cardiovascular protection, and detoxification. To address the industrial bottleneck of component degradation during processing, non-thermal technologies such as high hydrostatic pressure, ultrasound, and microencapsulation are discussed as stabilization solutions. Applications of wheat seedlings in functional foods, including sleep-aid supplements, fortified baked goods, and beverages, are also presented. Future research directions, such as precision breeding, multi-component synergy, and targeted product development, are outlined. This review focuses on two core issues: the efficient enrichment of bioactive compounds and the preservation of their stability during processing.

High-molecular-weight glutenin subunits (HMW-GSs) are key contributors to wheat dough viscoelasticity, with their conformational adaptability playing a vital role in gluten network formation. In this study, we employed a two-stage molecular dynamics simulation approach to investigate how acidic (pH 2) and alkaline (pH 12) conditions modulate the metastable states of three x-type HMW-GSs-Ax1, Bx7, and Dx2-and how these conformations respond to thermal aggregation. Results revealed clear subunit-specific differences in conformational plasticity. Under extreme pH conditions, Ax1 and Bx7 displayed greater RMSD fluctuations and broader radius of gyration distributions compared to Dx2, demonstrating enhanced structural flexibility. Hydrogen bond lifetime analysis indicated that Bx7 maintained longer-lived interactions across metastable states, implying greater internal stabilization. Upon thermal acceleration at 400 K, Ax1 rapidly progressed toward aggregation-prone conformations, whereas Dx2 preserved partial structural integrity, reflecting its superior thermal resilience. Covariance-based cross-correlation analysis further showed extensive domain decoupling in Ax1 and Bx7, in contrast to the more coordinated dynamics of Dx2. By linking metastable ensembles to aggregation behavior, this study provides mechanistic insights into gluten protein adaptability. These molecular insights may also serve as a reference for understanding hydration, gelation, and viscoelasticity of gluten networks in wheat-based food systems.

This study proposes moderate electric field (MEF)-assisted germination as a clean-label strategy for tailoring brown rice (BR) starch through biologically mediated modification. MEF-assisted germination accelerates starch degradation, resulting in 12.81% lower starch content than the control at 5 d of germination. Multi-scale structural analyses revealed a stage-dependent evolution of starch architecture. At the early stage (1 d), MEF-treated starch exhibited higher crystallinity (24.64%) than the control (24.02%), indicating enhanced structural ordering. This was accompanied by enrichment of short amylopectin chains and increased short-range molecular order. In contrast, at later stages (3-5 d), MEF accelerated structural disintegration, leading to reduced crystallinity, disruption of double helices, and fragmentation of starch granules. These structural changes were closely associated with functional properties. MEF decreased water solubility and swelling power at the early stage but significantly increased them at later stages. A similar biphasic pattern was observed in digestibility: at day 1, MEF reduced rapidly digestible starch (RDS) by 3.41% while increasing slowly digestible starch (SDS) and resistant starch (RS) by 2.76% and 0.64%, respectively, whereas the opposite trend occurred at later stages. Notably, direct MEF treatment of isolated starch caused negligible structural changes, whereas the α-amylase activity increased during MEF-assisted germination, suggesting that starch modification is likely related to enzyme-mediated hydrolysis. Overall, this study classified that MEF-assisted germination could regulate starch structure and functionality via enzyme-related pathways in a stage-dependent manner, providing a potential strategy for tailoring starch properties in whole-grain systems.

Gas-cell stability during dough fermentation depends on the ability of dough liquor (DL) proteins to adsorb and stabilize the air-water interface. However, the subunit-specific contributions of x-type high-molecular-weight glutenin subunits (HMW-GS) to this interfacial process remain unclear. Here, all-atom molecular dynamics (MD) simulations were used to examine the interfacial responses of Ax1, Bx7, and Dx2. Ax1 maintained relatively stable conformational features near the interface, Bx7 underwent moderate structural adjustments, whereas Dx2 exhibited pronounced interfacial sensitivity characterized by enhanced fluctuations and hydration-electrostatic redistribution. Consistent with these molecular differences, experimental analyses of DL extracted from fermented dough revealed that deletion of Dx2 markedly weakened interfacial performance, as reflected by reduced foaming stability, accelerated drainage, elevated surface tension, and lower interfacial viscoelasticity. These changes were accompanied by decreased ζ-potential and surface hydrophobicity, increased free -SH content, and reduced β-sheet structure, corresponding to diminished gas retention during fermentation. Together, the integrated simulation and experimental results demonstrate that subunit-dependent interfacial adaptability of x-type HMW-GS critically influences DL functionality and fermentation performance.

Propionibacterium freudenreichii has been widely used for industrial production and in situ fortification of vitamin B12 (VB12) in foods. This study systematically investigated the impacts of carbon/nitrogen sources, oxygen conditions, and cobalt availability on growth, metabolism, and VB12 biosynthesis in two P. freudenreichii strains (CICC 10019 and DSM 20271). Meanwhile, the composition of three types of VB12, namely methylcobalamin (MeCbl), adenosylcobalamin (AdoCbl) and hydroxocobalamin (OHCbl), during fermentation was also determined by an LC-MSMS method. The results showed carbon and nitrogen sources affect the yield and composition of VB12, but AdoCbl remained the predominant form in all conditions. Although P. freudenreichii is anaerobic, aeration can significantly increase the biomass and VB12 yield (especially AdoCbl) during incubation, accompanied by late-stage pH rebound. Cobalt was essential for VB12 biosynthesis, with maximal utilization efficiency at 0.1 mu g/mL CoCl2. CICC 10019 and DSM 20271 exhibited distinct responses during the logarithmic phase, revealing strain-specific regulatory differences. CICC 10019 upregulated propanoate metabolism genes (mutA/B) to enhance AdoCbl synthesis, while DSM 20271 downregulated corrin ring biosynthesis genes and cobalt incorporation enzymes, leading to relatively lower production of VB12. Both strains downregulated cobalt transporters (cbiM/N) under cobalt repletion, indicating adaptive uptake regulation. These findings provide critical insights for optimizing VB12 production and tailoring cobalamin profiles for fortification of this nutrient during food fermentation.

This study explores how germination duration affects the structure and gastrointestinal digestion of LAB-fermented soymilk gels. Soybeans germinated for 0-3 days were fermented, with their rheology, microstructure, intermolecular interactions, and in vitro digestion analyzed. Germination weakened gel viscoelasticity, increased pore size, and shifted the dominant forces from hydrogen bonds to hydrophobic interactions. This is likely due to the higher endogenous proteases and 7S/11S ratio. Germination-derived gels (especially 2-3 days) disintegrated faster in the stomach, released more soluble protein in the intestine, and produced more peptides. Peptidomic analysis revealed that 2-day germination (fermented soft gel, FSG) yielded the most bioactive peptides via targeted subunit degradation, while 3-day germination (fermented ultrasoft gel, FUG) caused over-hydrolysis, leading to loss of specific bioactive motifs. These findings demonstrate that controlled germination time (2-day) optimizes LAB-fermented gel structure to improve digestive efficiency and bioactive peptide release, providing a novel strategy for designing plant-based dairy.

Soybean isoflavones (SIs) are pivotal bioactive targets in the growing plant-based functional food market due to their selective estrogen receptor modulator (SERM)-like properties. However, a significant disparity remains between agricultural SI accumulation and ultimate physiological efficacy, constrained by the yield-metabolism genetic antagonism "linkage drag" and the "dual-cage" matrix entrapment barrier. This review establishes an operationalized seed-to-table framework to bridge these gaps through precise profile tailoring. We critically evaluate upstream genetic interventions (e.g., CRISPR/Cas9-mediated flux redirection) and downstream processing (e.g., biotransformation, physical-field intensification, and green solvent engineering). A novel, evidence-based key performance indicator (KPI) matrix is introduced to benchmark these technologies against standardized technology readiness levels (TRLs), economic burden (CapEx/OpEx), and systemic trade-offs. Crucially, we reframe SI bioavailability by accounting for Phase II metabolism, emphasizing that aglycone enrichment optimizes pharmacokinetic consistency and absorption kinetics rather than absolute systemic exposure. By integrating human clinical evidence and safety considerations for vulnerable populations (e.g., ER+ breast cancer survivors), this work bridges the persistent validation gap between laboratory innovation and clinical outcomes. Finally, we outline a paradigm shift toward AI-driven digital twins and host metabotyping to resolve the sensory-bioactivity paradox, providing a scientific roadmap for the rational design of next-generation isoflavone-fortified soy foods.

Three varieties of rice (Huaidao NO. 5, H5; Longgeng NO. 9, L9; Daohuaxiang NO. 2, D2) were cooked for varying durations (30-50 min), processed into prefabricated fried rice, and then subjected to long-term frozen storage (0-24 weeks) to investigate the mechanisms of quality change and structural evolution. Results indicated that cooking time significantly influenced rice quality, with H5 and L9 achieving optimal palatability (moderate hardness, high adhesiveness and maximum peak viscosity) at 40 min, while D2 peaked at 35 min. Electronic nose and GC-IMS analyses revealed that D2 possessed the most diverse volatile profile, particularly rich in aldehydes and unique ester markers. During frozen storage, starch retrogradation led to a progressive reduction in moisture content across all samples. By week 24, the hardness of H5, L9, and D2 increased by 1.44, 1.65, and 1.99 times, respectively, while chewiness increased by 2.81, 2.08, and 3.50 times. Scanning Electron Microscopy revealed a time-dependent microstructural transition from a dense, continuous gel matrix to a highly porous, sponge-like network. Concurrently, XRD and FTIR analyses confirmed that starch chains had reorganized into a crystalline structure. Overall, H5 exhibited superior freezing tolerance and stability, whereas D2 showed the most significant susceptibility to rapid retrogradation and textural degradation. These findings provide a theoretical basis for optimizing the processing and storage protocols of industrial prefabricated fried rice.

Plant-derived exosome-like vesicles (PDEVs) have emerged as promising oral delivery carriers, yet their application is severely limited by gastrointestinal instability and poor intestinal absorption. To overcome these barriers, we developed a biohybrid nanoplatform by sequentially coating wheat-seedling PDEVs with trimethyl chitosan (TMC) and carboxymethyl chitosan (CMC) via layer-by-layer self-assembly. This engineering strategy confers enhanced physicochemical stability, effectively protecting the payload against gastric degradation while enabling targeted intestinal release. Mechanistically, the multifunctional shell promotes transepithelial transport by enhancing cellular internalization and reversibly modulating epithelial tight junctions. Importantly, this improved delivery enables synergistic immuno-metabolic effects between the loaded α-lipoic acid (ALA) and the carrier's endogenous bioactives (GABA and folic acid). In a metabolic syndrome cell model, the nanoplatform achieved therapeutic efficacy superior to free drug combinations by suppressing pro-inflammatory cytokines and regulating lipid metabolism. Furthermore, in a high-fat diet-induced zebrafish model, the system significantly reversed abdominal lipid accumulation and alleviated systemic oxidative stress. Overall, this work provides a generalizable surface-engineering strategy for stabilizing natural vesicle carriers and supports their application in synergistic oral metabolic therapy.

To decipher how the gluten molecular matrix cooperates with micro-nano porous structures to modulate mechanical properties, we selected two wheat varieties for each of the three gluten strength levels (high, medium, and low). Through systematic analyses focusing on molecular conformation, network structure, and mechanical behavior, we developed a predictive model linking structural features to functional properties. The results showed that, compared with low-gluten samples, high-gluten samples exhibited significantly higher glutenin macropolymer content (41.8%) and glutenin/gliadin ratio (1.34). Moreover, high-gluten samples possessed a higher proportion of intermolecular β-sheet and a lower proportion of α-helix, with the SDS-insoluble fraction accounting for 36.6% versus 25.2% in low-gluten samples. The protein network area fraction of high-gluten samples was greater than that of low-gluten samples, and the pore size distribution was more uniform. However, differences in nanopore parameters among samples were limited. Mechanical testing revealed that high-gluten samples exhibited higher storage modulus, frequency sensitivity, maximum tensile resistance, and R/E ratio, and their power-law elastic strength (Af) was also significantly superior to that of other samples. Based on multi-scale structural factors, multiple linear regression achieved the best prediction for maximum tensile resistance (R), while partial least squares regression performed best for predicting R/E. However, both models showed relatively lower predictive accuracy for Af and Z values. The multi-scale structure–property model established in this study can effectively predict the processing characteristics of gluten, providing a theoretical basis for flour quality evaluation and product formulation optimization.

Arabinoxylan (AX) from wheat bran is an effective antifreeze polysaccharide, but its structure–activity relationship in ionic environments remains poorly understood. In this study, representative Hofmeister salts (Na2SO4, NaCl, NaNO3, KCl, and MgCl2) were introduced into AX solutions to elucidate the ion-specific regulation of AX antifreeze activity. Heterogeneous ice nucleation, single ice-crystal growth, and ice recrystallization inhibition analyses demonstrated that ions significantly affect AX activity, with anions playing a more pronounced role than cations. At 0.05 M, Na2SO4 markedly enhanced the antifreeze activity of AX, whereas NaCl and NaNO3 showed limited or inhibitory effects. Structural analyses revealed that AX adopts a flexible random-coil conformation in solution, while sulfate ions promoted a more compact molecular organization. Integrated mechanistic analyses suggested that ion-induced conformational rearrangement and hydration regulation collectively modulate AX–associated hydration environments and enhance its ice-affinity. Specifically, SO42-–induced AX compaction was accompanied by altered interfacial hydrogen-bonding states, stabilized local hydration interactions, and modulation of long-range hydrogen-bond network dynamics. These findings establish an ion-specific conformation–hydration coupling mechanism for understanding and regulating polysaccharide antifreeze activity.

The rational design of antifreeze peptides remains a significant challenge. In this study, we developed a series of short peptides with antifreeze activity: G-1 (GPACCCGPA), G-2 (GPACCCGPA)2, and G-3 (GPACCCGPA)3, by mimicking the repetitive sequences and structural motifs of natural antifreeze proteins (AFPs). Notably, antifreeze activity was enhanced with increasing peptide length and molecular weight, with G-3 exhibiting the most pronounced ice recrystallization inhibition. Systematic assays, including splat cooling, sucrose sandwich, and nanoliter osmometry, revealed that peptide elongation markedly suppressed ice growth rates without inducing thermal hysteresis (TH). Structural analyses demonstrated that increased peptide length enhanced α-helical content and promoted the formation of ordered nanostructures with moderate aggregation, enabling multiple ice-binding motifs to interact directly with the ice-water interface. The combined results of water spectral analysis and molecular dynamics simulations further revealed that peptides modulated the organization of surrounding water clusters by increasing local hydrogen-bond ordering while disrupting long-range hydrogen-bond dynamics. This dual modulation weakened the integrity of the extended hydrogen-bond network in bulk water and delayed the transition into a highly ordered crystalline ice phase. Mechanistically, G-3 achieves dual functionality by coupling efficient initial ice templating with strong inhibition of crystal growth, providing a clear rationale for its superior activity. Furthermore, the peptides exhibited excellent biocompatibility and significantly improved post-thaw cell viability. Overall, the design strategy proposed in this study highlights the potential of modular peptide engineering for developing biocompatible, tunable cryoprotectants, offering mechanistic insights into peptide-water interactions and clarifying the molecular basis of peptide-mediated antifreeze activity.