Antifreeze Polysaccharides from Wheat Bran: The Structural Characterization and Antifreeze Mechanism.

Exploring a novel natural cryoprotectant and understanding its antifreeze mechanism allows the rational design of future sustainable antifreeze analogues. In this study, various antifreeze polysaccharides were isolated from wheat bran, and the antifreeze activity was comparatively studied in relation to the molecular structure. The antifreeze mechanism was further revealed based on the interactions of polysaccharides and water molecules through dynamic simulation analysis. The antifreeze polysaccharides showed distinct ice recrystallization inhibition activity, and structural analysis suggested that the polysaccharides were arabinoxylan, featuring a xylan backbone with a majority of Araf and minor fractions of Manp, Galp, and Glcp involved in the side chain. The antifreeze arabinoxylan, characterized by lower molecular weight, less branching, and more flexible conformation, could weaken the hydrogen bonding of the surrounding water molecules more evidently, thus retarding the transformation of water molecules into the ordered ice structure.

Enhancing the technofunctionality of γ-aminobutyric acid enriched germinated wheat by modification of arabinoxylan, gluten proteins and liquid lamella of dough.

To enhance the technofunctionality of germinated wheat enriched with γ-aminobutyric acid, xylanase (Xyn) and glucose oxidase (Gox) were incorporated with emphasis on modifying the key components. Combination of Xyn and Gox enhanced steamed bread quality with optimum loaf volume and textural property. Continuous and dense gluten network was facilitated and improved viscoelasticity of dough. Water solubility of arabinoxylan (AX) enhanced with Xyn and the molecular weight was more homogeneous distributed throughout bread making process with Xyn and Gox. Polymerization behavior of α-/γ-gliadin and glutenin was suppressed in steamed bread, while incorporation of AX to insoluble proteins was enhanced by enzymes. In addition, the promoted formation of high molecular weight glycoprotein in the liquid lamella of dough enhanced the thermal stability of foams and contribute to superior quality of steamed bread. Results demonstrated that germinated wheat could be exploited as a functional ingredient with desirable technofunctionality by modification of the components.

Spermidine alleviates oxidative damage and enhances phenolic compounds accumulation in barley seedlings under UV-B stress.

BACKGROUND: Ultraviolet B (UV-B) radiation can enhance the accumulation of phenolic compounds (PCs) in barley seedling, although this may result in severe oxidative damage. In the present study, the role of spermidine in alleviating oxidative damage and regulating synthesis of PCs in barley seedlings under UV-B stress was investigated. RESULTS: Exogenous spermidine increased the length and fresh weight as well as PCs contents of barley seedlings under UV-B stress. Application of dicyclohexylamine, an inhibitor of endogenous spermidine synthesis, significantly inhibited the growth and PC accumulation of barley seedlings under UV-B stress, although this inhibitory effect can be alleviated by exogenous spermidine. Exogenous spermidine increased the contents of vanillic acid, syringic acid, protocatechuic acid and p-coumaric acid in barley seedlings under UV-B stress by 20-200% through enhancing the activities of enzymes related to synthesis of these acids. In addition, exogenous spermidine enhanced activities and gene expression of antioxidant enzymes in barley seedlings under UV-B stress, including peroxidase, glutathione reductase and glutathione S-transferase. CONCLUSION: Spermidine can alleviate oxidative damage of barley seedlings under UV-B stress and enhance the accumulation of PCs. © 2022 Society of Chemical Industry.

Impact of Different Frozen Dough Technology on the Quality and Gluten Structure of Steamed Buns.

To advance the industrialization production of steamed buns, the current study explored the freeze-stability of unfermented, pre-fermented and par-steamed frozen dough. The results showed that the steamed bun made from unfermented dough with 2.0% yeast, the pre-fermented dough with a pre-fermented time of 30 min and the par-steamed dough with a pre-steamed time of 15 min showed the best sensory properties quality upon frozen storage. The gassing power of un- and pre-fermented dough gradually decreased, and dough with longer pre-fermented time exhibited more evident loss of gassing power. Freeze-induced depolymerization of gluten protein was the least distinct in the par-steamed dough, followed by the pre- and un-fermented dough, which was probably related to the superior freeze stability of glutenin-gliadin macro-crosslinks upon the pre-steaming stage. The surface hydrophobicity of gluten proteins of frozen dough decreased during the initial storage and was enhanced subsequently, which was related with the combined effects of the unfolding and synchronous aggregation induced by freezing and steaming, respectively. Moreover, the surface hydrophobicity of gluten in par-steamed frozen dough and steamed buns was more resistant to frozen storage, which was probably attributed to the established stable structure during the pre-steaming process.

Impact of Wheat Arabinoxylan with Defined Substitution Patterns on the Heat-Induced Polymerization Behavior of Gluten.

To further depict the interaction mechanism of wheat arabinoxylan (AX) and gluten proteins upon thermal processing, AX was enzymatically tailored with defined substitution patterns and the impact on the heat-induced polymerization behavior of gluten was comparatively studied. The results showed that tailormade AX promoted the formation of glutenin-glutenin and glutenin-gliadin macrocrosslinks upon heating, with the optimal effect detected for AX depleted of Araf of disubstituted Xylp. The tailormade AX, especially AX depleted of monosubstituted Xylp, facilitated the polymerization ability of α-gliadin into glutenin compared with untailored AX. The unfolding process of gluten was partially impeded by AX upon heating, while the tailormade AX promoted the unfolding process. AX could bury Trp and Tyr upon polymerization of glutenin and gliadin and induced the change of the disulfide bridge conformation to a less-stable state, while the effect was alleviated with tailormade AX. The enhanced polymerization with tailormade AX strengthened the gluten network and induced more heterogeneously distributed large protein aggregates.

UV-B- triggered H2O2 production mediates isoflavones synthesis in germinated soybean.

In this study, the functions of Hydrogen peroxide (H2O2) on the synthesis of isoflavones in germinated soybean under UV-B radiation were investigated. Results showed that the activity, gene, and protein expression of NADPH oxidase were up-regulated by 1.46, 6.92, and 1.34 times with UV-B radiation, while endogenous H2O2 content was also significantly increased. UV-B radiation and exogenous H2O2 treatment significantly increased the activities, gene and protein expression of phenylalanine ammonia lyase (PAL), chalcone synthase (CHS), and isoflavone synthase (IFS) involved in isoflavones synthesis, and there was a synergistic effect with combining treatment. However, these up-regulation effects were suppressed by the supplementary diphenylene iodonium (DPI), which is the inhibitor of NADPH oxidase. Interestingly, the inhibition effect was largely reversed by exogenous H2O2, indicating that H2O2 was indispensable in regulating the isoflavones synthesis in germinated soybeans under UV-B radiation. Overall, H2O2 is an essential signaling molecule, mediating UV-B-induced isoflavone accumulation.

Determination of glucosinolates in rapeseed meal and their degradation by myrosinase from rapeseed sprouts.

The utilization of rapeseed meal in food is limited due to its abundant glucosinolates (GLs). In this study, an LC-MS/MS method for GLs determination in rapeseed meal was developed. Then, the degradation of GLs using rapeseed sprouts derived myrosinase (MYR) was investigated. Results showed that 11 kinds of GLs were identified in rapeseed meal. The LC-MS/MS method had a high linearity (R2 greater than 0.9999), repeatability (RSD < 5%) and recovery rate (92%-102%). The optimum condition for hydrolyzing GLs in rapeseed meal was reacting for 4 h with the addition of 2236.35 U/g MYR, 9.63 µg/g ascorbic acid and 26.68 µg/g EDTA. Under this condition, more than 80% of GLs were degraded and the yields of isothiocyanates and oxazolidinone-2-thione were 859.30 µg/g and 685.59 µg/g, respectively. To conclude, this study reported a reliable method for GLs determination and an effective way to degrade GLs in rapeseed meal.

Tailormade Wheat Arabinoxylan Reveals the Role of Substitution in Regulating Gelatinization and Retrogradation Behavior of Wheat Starch.

To elucidate the role of substitution of arabinoxylan (AX) in the characteristics of wheat starch, this study prepared AX with a well-defined structure by targeted enzymatic hydrolysis and comparatively investigated the effects of AX with different degrees of substitution on gelatinization and retrogradation behavior of starch. Removal of major arabinofuranosyl (Araf) of mono- or disubstituted xylopyranosyl (Xylp) of both low-molecular-weight (Mw: 62.5 kDa, Araf/Xylp: 0.61) and high-molecular-weight AX (Mw: 401.2 kDa, Araf/Xylp: 0.61) reversed the decreased gelatinization viscosity and recrystallization of amylose induced by AX to a similar extent. Upon retrogradation for 30 days, the Araf of mono- and disubstituted Xylp contributed to the water distribution and the effect depended on the molecular chain length. The C3-linked Araf of disubstituted Xylp was more involved in prohibiting the retardation of recrystallization of amylopectin, while the presence of Araf of monosubstituted Xylp might hinder the interactions between AX and amylopectin.

Red light enhances folate accumulation in wheat seedlings.

Red, white, blue, green, and yellow lights were applied to investigate their effects on folate accumulation in wheat seedlings. The different lights, especially red light, significantly increased the total folate content. Total folate showed maximum accumulation under 30 µmol/(m2·s) of red light, with an increase of 24% compared with the control (darkness). 5-Methyl-tetrahydrofolate (5-CH3-THF) was the dominant folate component, and was significantly increased by red light irradiation. In addition, under red light, the folate content of leaves was higher and more sensitive to light than that of endosperm or roots. Red light up-regulated the expression of guanosine triphosphate (GTP) cyclohydrolase 1 (GCH1) and aminodeoxychorismate synthase(ADCS), enhanced the activity of GCH1 and ADCS, and increased the content of precursors of folate synthesis, including pterin and p-aminobenzoic acid (pABA). Hence, the increased folate accumulation promoted by light could be attributed to the increased content of folate synthesis precursors, the activity of key enzymes, and related gene expression.

Effect of γ-aminobutyric Acid on Phenolics Metabolism in Barley Seedlings under Low NaCl Treatment.

It has been revealed that high NaCl stress (>60 mmol L-1) induced phenolics accumulation in barley seedlings, with γ-aminobutyric acid (GABA) playing a key role. Interestingly, low NaCl stimulus (20 mmol L-1) enhancing phenolics synthesis and growth of barley seedlings was also reported recently. Hence, exogenous GABA and its bio-synthesis inhibitor 3-mercaptopropionic acid (3-MP) were applied to reveal the mechanism of GABA regulating phenolics metabolism in barley seedlings treated with 20 mmol L-1 NaCl. The contents of total phenolics and flavonoids significantly increased by 11.64% and 14.52% under NaCl, respectively. The addition of GABA further increased phenolics and flavonoids contents, especially for gallic acid, protocatechuic acid, caffeic acid, and quercetin, compared with NaCl treatment. Simultaneously, GABA increased the activities and mRNA levels of phenylalanine ammonia lyase (PAL), cinnamic acid 4-hydroxylase (C4H), and 4-coumalyl CoA ligase (4CL). The addition of 3-MP suppressed the above effects, except for increasing the protein levels of PAL, C4H, and 4CL. Low concentration of NaCl not only promoted growth, but also stimulated endogenous GABA metabolism to affect key enzymes activities and mRNA levels for phenolics synthesis in barley seedlings.

GABA Regulates Phenolics Accumulation in Soybean Sprouts under NaCl Stress.

NaCl stress causes oxidative stress in plants; γ-aminobutyric acid (GABA) could alleviate such abiotic stress by enhancing the synthesis of phenolics, but the underlying mechanism is not clear. We investigated the effects of GABA on phenolics accumulation in soybean sprouts under NaCl stress by measuring changes in the content of physiological biochemicals and phenolic substances, in the activity and gene expression of key enzymes, and in antioxidant capacity. GABA reduced the oxidative damage in soybean sprouts caused by NaCl stress and enhanced the content of total phenolics, phenolic acids, and isoflavones by 16.58%, 22.47%, and 3.75%, respectively. It also increased the activities and expression of phenylalanine ammonia lyase, cinnamic acid 4-hydroxylase, and 4-coumarate coenzyme A ligase. Furthermore, GABA increased the activity of antioxidant enzymes and the antioxidant capacity. These events were inhibited by 3-mercaptopropionate (an inhibitor for GABA synthesis), indicating that GABA mediated phenolics accumulation and antioxidant system enhancement in soybean sprouts under NaCl stress.

NaCl treatment on physio-biochemical metabolism and phenolics accumulation in barley seedlings.

Phenolics are important secondary metabolites in plants with strong antioxidant effects. Seeds germination and exogenous stimulation could activate endogenous enzymes to enhance the content of phenolic acids and flavonoids. Barley seeds geminated under NaCl (1-20 mM) treatment to evaluate the accumulation of phenolics in this study. Results showed that NaCl treatment significantly enhanced the growth of seedlings, especially bud length. NaCl treatment up-regulated genes and proteins expression of phenylalanine ammonia lyase (PAL), cinnamate-4-hydroxylase (C4H) and 4-coumarate-CoA ligase (4CL), resulting in the enhancement of their activities. As a result, phenolic acids and flavonoids contents increased by 11.19% and 32.54%, respectively, in which gallic acid, protocatechuic, fisetin, myricetin and quercetin were affected mostly. Moreover, NaCl treatment enhanced 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging capacity. Hence, NaCl stimulated the synthesis of phenolic components via enhancing gene, protein expression and the activity of key enzymes.

Impact of water extractable arabinoxylan with different molecular weight on the gelatinization and retrogradation behavior of wheat starch.

Water-extractable arabinoxylan (WEAX) could effectively improve the cereal food quality, while its regulatory effect on wheat starch properties has yet to be well-understood. This study selected the WEAX with different molecular weight (Mw) but same branched degree, and comparatively investigated their effects on the gelatinization and retrogradation behavior of wheat starch. The decreased degree of swelling power, solubility and peak viscosity suggested that low Mw WEAX (L-WEAX) could hinder starch gelatinization more evidently compared with high Mw WEAX (H-WEAX), due to the pronounced inhibition effect on amylose leaching and amylose-lipid complex formation. L-WEAX suppressed the recrystallization of amylose and thus the short-term retrogradation. However, H-WEAX mainly retarded the recrystallization of amylopectin, exerting a more significant inhibition effect on the long-term retrogradation. This study could provide a theoretical basis for enhancing the quality and extending the shelf life of starchy foods by selecting the optimum structure of WEAX.

Water-Extractable Arabinoxylan-Induced Changes in the Conformation and Polymerization Behavior of Gluten upon Thermal Treatment.

Interactions between gluten proteins and water-extractable arabinoxylan (WEAX) during the heating stage are crucial for the organoleptic quality of high-fiber cereal products. To reveal the molecular mechanism of WEAX on gluten characteristic upon heating, the current study comparatively investigated the effects of WEAX with different molecular weights (Mw) on the heat-evoked conformational variation and polymerization behavior of gluten. Results showed that WEAX, especially low Mw WEAX (L-WEAX), facilitated the polymerization ability of α-/γ-gliadins into glutenins, whereas high Mw WEAX (H-WEAX) reduced the polymerizing temperature of glutenin and gliadin. L-WEAX could develop more hydrogen bonds with tyrosine of gluten and stabilize the secondary structure more evidently than H-WEAX upon heating. Compared with disulfide bridge formation, hydrophobic interactions were not the driving force involved in the heat-induced polymerization behavior affected by WEAX. WEAX evoked the reinforced glutenin network and heterogeneous distribution of gliadin, with a more uniform molecular surface developed for gluten.

The mechanism of freeze-thawing induced accumulation of γ-aminobutyric acid in germinated soybean.

BACKGROUND: γ-Aminobutyric acid (GABA) is a non-protein amino acid with several functions in the human body. Although freeze-thawing could effectively accumulate GABA in soybean sprouts, the mechanism has not been revealed. The mechanism by which freeze-thawing enhances GABA accumulation in germinated soybean was revealed by evaluating GABA content, the activity of related synthesis enzymes, and the microstructure of the tissues and cells of sprouts. The germinated soybeans were treated at different temperatures (from -196 °C to 25 °C) for 12 h and then thawed at 25 °C for 6 h. RESULTS: The results showed that GABA content in frozen soybean sprouts did not change significantly before thawing. After thawing, the GABA content of sprouts increased by 83.9% and 82.9% when treated by liquid nitrogen flash freeze at - 80 °C for 12 h compared with the control (4 °C treatment for 12 h). The results indicated that GABA formation mainly occurred during thawing. However, glutamate decarboxylase (GAD), diamine oxidase (DAO), and aminoaldehyde dehydrogenase (AMADH) activity decreased during thawing. Based on the malonaldehyde (MDA) content and microstructure of sprouts, it was suggested that freezing at lower temperatures (< -20 °C) maintained the integrity of the cell structure, while the tissues and cell membranes were broken during thawing. CONCLUSION: These results could provide evidence for the hypothesis that GABA formation resulted from full contact between enzymes and substrates during thawing, rather than the contribution of higher enzyme activity. © 2019 Society of Chemical Industry.

Spermidine improves antioxidant activity and energy metabolism in mung bean sprouts.

The effects of exogenous spermidine and dicyclohexylamine (DCHA, spermidine synthesis inhibitor) on the antioxidative system and energy status of germinating mung bean were investigated. Results showed that exogenous spermidine increased the content of total phenolic and ascorbic acid and the antioxidative activity, but reduced activities and gene expressions of peroxidase (POD) and catalase (CAT). These changes might be explained by increased H2O2 content and activities of succinic dehydrogenase (SDH), adenosine triphosphatases (ATPases), and cytochrome c oxidase (CCO), resulting in higher adenosine triphosphate (ATP) and energy charge (EC). Interestingly, spermidine down-regulated expressions of SDH, H+-ATPase, Ca2+-ATPase and CCO whilst DCHA reduced energy metabolism and induced the opposite effects to spermidine, except for ascorbic acid content. Inhibition was reversed by exogenous spermidine. In conclusion, spermidine induced the accumulation of H2O2, enhanced the antioxidative system and improved the energy metabolism to enhance the functional quality of mung bean sprouts.

Preparative isolation and purification of B-type fumonisins by using macroporous resin column and high-speed countercurrent chromatography.

B-type fumonisins (FBs) are water-soluble mycotoxins produced by Fusarium species, which are mainly found in maize products and threaten food safety. Toxicological studies and quantitative determinations of fumonisins require large amounts of pure toxins, and their high prices limit progress in FBs research. In this study, we used a macroporous resin column combined with high-speed countercurrent chromatography to separate large quantities of FBs. A fermented rice culture was extracted with 75% methanol. The dynamic adsorption capacity of FBs on XAD-2 resin was 27.5 mg/g resin at 25°C, pH 4.0, and then the FBs were desorbed with 60% methanol. The crude FBs were further purified using a biphasic system consisting of n-heptane/n-butanol/methanol/water (2:4:1:4, v/v/v/v). The method yielded 1.55 g of FB1 and 0.55 g of FB3 with purities of 96.8% and 95.6%, respectively, from 1 kg of rice culture, and the final overall yield of FBs was 74.8%.

Ca2+ involved in GABA signal transduction for phenolics accumulation in germinated hulless barley under NaCl stress.

In this study, in order to investigate the role of Ca2+ in GABA signal transduction involved in phenolics accumulation in barley seedlings under NaCl stress, the seedlings were treated with exogenous GABA and its synthesis inhibitor, 3-mercaplopropionic acid (3-MP), as well as Ca2+ channel blockers La3+, Ca2+ chelator EGTA, and Ca2+ release channel inhibitor 2-aminoethoxydiphenyl borate (2-APB). The results showed that GABA significantly enhanced phenolics, calcium and calmodulin content. It also induced Ca2+ influx in barley root tips cells, and altered the distribution of Ca2+, making calcium precipitates more uniform and intensive. While, 3-MP treatment led to opposite changes, which suggested that GABA was essential for calcium content increase. In addition, accumulation of phenolics was inhibited by LaCl3, EGTA and 2-APB treatments, and this inhibition could be alleviated partly by exogenous GABA. Taken together, Ca2+ was involved in GABA signal transduction for phenolics accumulation in barley seedlings under NaCl stress.

AMADH inhibitor optimization and its effects on GABA accumulation in soybean sprouts under NaCl-CaCl2 treatment.

ABSTRACT: 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide (EDC) was the suitable inhibitor for aminoaldehyde dehydrogenase (AMADH) compared with N-ethylmaleimide (NEE) and iodoacetamide (IAM). EDC exhibited the most obvious inhibition effect on AMADH activity, while its inhibition on glutamate decarboxylase (GAD) was insignificant. Compared with the control, AMADH activity reduced by 70.4% with 0.5 mM EDC, and γ-aminobutyric acid (GABA) content declined by 44.3% in soybean sprouts at 4 days of germination. AMADH activity reduced by 80.62, 67.61 and 72.02% in the 4-day sprouts with 1 mM EDC under NaCl, CaCl2 and NaCl + CaCl2 treatment, respectively, and GABA content decreased by 43.56, 38.84 and 35.53%, respectively. EDC is a proper inhibitor for AMADH and it could be used to quantify the contribution of polyamine degradation pathway on GABA formation. In soybean sprouts, the presence of CaCl2 under NaCl stress decreased the contribution of polyamine degradation pathway on GABA accumulation.

iTRAQ-based proteomic analysis reveals changes in response to sodium nitroprusside treatment in soybean sprouts.

In recent years, nitric oxide (NO) has been considered a plant signaling compound involved in antioxidant systems and flavonoid production enhancement. Nevertheless, its mechanism of action, from the perspective of protein expression, remains largely unknown. In this study, isobaric tags for relative and absolute quantitation (iTRAQ) was employed to investigate NO donor sodium nitroprusside treatment-induced proteomic changes in soybean sprouts. Among the 3033 proteins identified, compared with the control, sodium nitroprusside treatment up- and down-regulated 256 proteins. These proteins were involved in antioxidant system pathways, such as the thioredoxin, superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), glutathione reductase (GR), glutathione S-transferase (GST), ascorbate peroxidase (APX), monodehydroascorbate reductase (MDAR) and lipoxygenase (LOX) pathways, including allene oxide synthase and lipoxygenase. In addition, heat shock proteins (HSPs) and flavonoid biosynthetic proteins, such as cinnamate 4-hydroxylase, chalcone isomerase, chalcone synthase, isoflavone synthase and isoflavone reductase, were also modulated in response to sodium nitroprusside treatment.