Investigating the Mechanism of Microwave-Assisted Enzymolysis Synergized with Magnetic Bead Adsorption for Reducing Ovalbumin Allergenicity through Biomass Spectrometry Analysis.

This study found that, after microwave treatment at 560 W for 30 s, alkaline protease enzymolysis significantly reduced the allergenicity of ovalbumin (OVA). Furthermore, specific adsorption of allergenic anti-enzyme hydrolyzed peptides in the enzymatic products by immunoglobulin G (IgG) bound to magnetic bead further decreased the allergenicity of OVA. The results indicated that microwave treatment disrupts the structure of OVA, increasing the accessibility of OVA to the alkaline protease. A comparison between 17 IgG-binding epitopes identified through high-performance liquid chromatography-higher energy collisional dissociation-tandem mass spectrometry and previously reported immunoglobulin E (IgE)-binding epitopes revealed a complete overlap in binding epitopes at amino acids (AA)125-135, AA151-158, AA357-366, and AA373-381. Additionally, partial overlap was observed at positions AA41-59, AA243-252, and AA320-340. Consequently, these binding epitopes were likely pivotal in eliciting the allergic reaction to OVA, warranting specific attention in future studies. In conclusion, microwave-assisted enzymolysis synergized with magnetic bead adsorption provides an effective method to reduce the allergenicity of OVA.

Characterization and in vitro digestibility of soybean tofu: Influence of the different kinds of coagulant.

This study explored the effect of diverse coagulants (glucono-δ-lactone (GDL), gypsum (GYP), microbial transglutaminase (MTGase), and white vinegar (WVG)) on microstructure, quality, and digestion properties of tofu. The four kinds of tofu were significantly different in their structure, composition, and digestibility. Tofu coagulated with MTGase had the highest springiness and cohesiveness while GDL tofu had the highest enthalpy (6.54 J/g). However, the WVG and GYP groups outperformed others in terms of thermodynamic, and digestion properties. The WVG group exhibited the highest nitrogen release (84.3%), water content, denaturation temperature, and the highest free-SH content but the lowest S-S content. Compared to WVG, the GYP group had the highest ash content, hardness, and chewiness. Results demonstrated that the tofu prepared by WVG and GYP show high digestibility. Meanwhile, the former has better thermal properties and the latter has better texture properties.

Insight into the Mechanism Underlying the Reduction of Digestibility and IgG/IgE Binding Ability in Ovalbumin during Different High-Temperature Conduction Modes-Induced Glycation.

Effects of different high-temperature conduction modes [high-temperature air conduction (HAC), high-temperature contact conduction (HCC), high-temperature steam conduction (HSC)]-induced glycation on the digestibility and IgG/IgE-binding ability of ovalbumin (OVA) were studied and the mechanisms were investigated. The conformation in OVA-HSC showed minimal structural changes based on circular dichroism, fluorescence, and ultraviolet spectroscopy. The degree of hydrolysis analysis indicated that glycated OVA was more resistant to digestive enzymes. Liquid chromatography-Orbitrap mass spectrometry identified 11, 14, and 15 glycation sites in OVA-HAC, OVA-HCC, and OVA-HSC, respectively. The IgG/IgE-binding ability of OVA was reduced during glycation and digestion, and the interactions among glycation, allergenicity, and digestibility were further investigated. Glycation sites masked the IgG/IgE epitopes resulting in a reduction in allergenicity. Digestion enzymes destroyed the IgG/IgE epitopes thus reducing allergenicity. Meanwhile, the glycation site in proximity to the digestion site of pepsin was observed to cause a reduction in digestibility.

Peptidomics Screening and Molecular Docking with Umami Receptors T1R1/T1R3 of Novel Umami Peptides from Oyster (Crassostrea gigas) Hydrolysates.

In this study, novel umami peptides were prepared from oyster (Crassostrea gigas) hydrolysates, and their umami mechanisms were investigated. Umami fractions G2 and G3 were isolated by gel filtration chromatography (GFC) and sensory evaluation. The umami scores of the G2 and G3 fractions were 7.8 ± 0.12 and 7.5 ± 0.18, respectively. 36 potential umami peptides with molecular weights below 1500 Da, E and D accounting for >30% of the peptides and iUmami-SCM > 588 were screened by peptidomics. Peptide source analysis revealed that myosin, paramyosin, and sarcoplasmic were the major precursor proteins for these peptides. The electronic tongue results demonstrated that the synthetic peptides DPNDPDMKY and NARIEELEEE possessed an umami characteristic, whereas SIEDVEESRNK and ISIEDVEESRNK possessed a saltiness characteristic. Additionally, molecular docking results indicated that the umami peptide (DPNDPDMKY, NARIEELEEE, SIEDVEESRNK, and ISIEDVEESRNK) binds to H145, S276, H388, T305, Y218, D216, and Q389 residues in the T1R3 taste receptor via a conventional hydrogen bond and a carbon-hydrogen bond. This research provides a new strategy for the screening of umami peptides.

Comparison of ovalbumin glycation induced by high-temperature steaming and high-temperature baking: A study combining conventional spectroscopy with high-resolution mass spectrometry.

High-temperature steaming (HTS) and high-temperature baking (HTB)-induced ovalbumin (OVA)-glucose glycation (140 °C, 1-3 min) were compared, and the different mechanisms were evaluated by changes in protein conformation, glycation sites and average degree of substitution per peptide molecule (DSP) values as well as the antioxidant activity of glycated OVA. Conventional spectroscopic results suggested that in comparison with HTB, HTS promoted protein expansion, increased ß-sheet content and made OVA structure more orderly. Liquid chromatography-high resolution mass spectrometry (LC-HRMS) analysis showed that 10 glycation sites were found under HTB, while 4 new glycation sites R111, R200, R219 and K323 appeared under HTS, and 2 of them (R219 and K323) were located in internal ß-sheet chains. The antioxidant activities of glycated OVA increased with increasing treatment time, and HTS showed stronger enhancement effect than HTB. Furthermore, the DSP values were generally higher under HTS than HTB. Compared with HTB, HTS with high penetrability could enhance the change of OVA primary structure and spatial conformation, making the protein structure more unfolded and stable, leading to more protein-sugar collisions occurred in inner OVA molecular and significantly promoted glycation. In conclusion, HTS is a promising method for high-temperature short-time glycation reaction, with drastically increasing the protein antioxidant activities.

Effects of different high-temperature conduction modes on the ovalbumin-glucose model: AGEs production and regulation of glycated ovalbumin on gut microbiota.

Food high-temperature processing frequently induces the production of advanced glycation end products (AGEs) in the food industry. In this study, the effects of three high-temperature conduction modes on the AGEs production derived from ovalbumin (OVA)-glucose model and the regulation of glycated OVA on gut microbiota were investigated. The peak time of OVA shifted maximally from 13.72 to 13.57 due to the rise in molecular weight, confirming successful coupling between OVA and glucose. The inhibition of superheated steam (SS) on AGEs was observed, with the sample treated by SS showing the lowest content among glycated OVA groups. The analysis revealed an increase in AGEs during digestion and a decrease in fermentation, suggesting the release during digestion and the availability by intestinal flora. Furthermore, an expansion of Bifidobacterium and Lactobacillus, and the inhibition of Desulfovibrio and Escherichia-Shigella were observed, indicating the prebiotic activity of glycated OVA and its potential to improve intestinal health. These results provide valuable information for controlling high-temperature processing to inhibit AGEs formation and highlight the positive effects of glycated proteins on intestinal health.

Effect of purple red rice bran anthocyanins on pasting, rheological and gelling properties of rice starch.

Purple red rice bran (PRRBA), a by-product of the rice polishing process, is frequently thrown away, resulting in a waste of resources. This study investigated the effects of PRRBA on the pasting, rheological property, chemical structure, microstructure, and water migration of rice starch. The results demonstrated that the peak viscosity (PV), final viscosity (FV), and activation energy (retrogradation energy) of rice starch were all decreased by a dose of PRRBA. Furthermore, the gel strength and hardness of rice starch were positively correlated with the addition of PRRBA. Rice starchs particle size distribution can be improved by PRRBA, which may be a result of the non-covalent bonds that exist between PRRBA and rice starch. The addition of PRRBA resulted in a decrease in the spin relaxation time (T2) of rice starch, from 259.7 to 143.6 ms. This can be attributed to that PRRBA improved the water-holding capacity of rice starch. These results could contribute to the development of high-value-added products of PRRBA and facilitate the application of anthocyanins in starch-based foods.

Effects of Superheated Steam Treatment on the Allergenicity and Structure of Chicken Egg Ovomucoid.

The aim of this study was to explore the effects of an emerging and efficient heating technology, superheated steam (SS), on the allergenicity and molecular structure of ovomucoid (OVM). OVM was treated with 120-200 °C of SS for 2 to 10 min. The allergenicity (IgG/IgE binding abilities and cell degranulation assay) and molecular structure (main functional groups and amino acids modification) changes were investigated. The IgG-binding ability of OVM decreased and the releases of ß-hex and TNF-γ were inhibited after SS treatment, indicating that the protein allergenicity was reduced. Significant increases in oxidation degree, free SH content and surface hydrophobicity were observed in SS-treated OVM. The protein dimer and trimer appeared after SS treatment. Meanwhile, obvious changes occurred in the primary structure. Specifically, serine can be readily modified by obtaining functional groups from other modification sites during SS treatment. Moreover, the natural OVM structure which showed resistance to trypsin digestion was disrupted, leading to increased protein digestibility. In conclusion, SS-induced OVM aggregation, functional groups and amino acids modifications as well as protein structure alteration led to reduced allergenicity and increased digestibility.

Preparation, characterization, antioxidant activity and protective effect against cellular oxidative stress of phosphorylated polysaccharide from Cyclocarya paliurus.

In this study, a phosphorylated derivative of Cyclocarya paliurus polysaccharide (CP) was synthesized using sodium tripolyphosphate and sodium trimetaphosphate. Antioxidant activities and cytoprotective effects of unmodified polysaccharide and phosphorylated derivatives were investigated employing various in vitro systems. Results showed that phosphorylated modification caused a significant change in chemical composition and the apparent structure of CP. Phosphorylated Cyclocarya paliurus polysaccharide (P-CP) showed excellent scavenging capacities against DPPH and hydroxyl radical in a dose-dependent manner. P-CP provided a more significant protective effect against hydrogen peroxide-induced oxidative stress in RAW264.7 cells by a compositive oxidation defense mechanism than CP. P-CP could reduce oxidative stress by significantly enhancing the contents of antioxidant enzyme superoxide dismutase (SOD) and decreasing malondialdehyde (MDA) content in oxidative damaged cells, in addition to scavenging ROS directly and regulating cell cycle distribution, reducing apoptosis capacity and improving cell viability, consequently achieving the intracellular antioxidant activity. This study suggested that phosphorylated modification is an effective method to improve the antioxidant activities of CP, so as to expand the development and application of CP.

Sulfated modification enhanced the antioxidant activity of Mesona chinensis Benth polysaccharide and its protective effect on cellular oxidative stress.

The effect of sulfated modification on the physicochemical properties and antioxidant activities of polysaccharide (MP) from Mesona chinensis Benth were investigated by chemical methods, Fourier transform infrared spectroscopy, high performance anion exchange chromatography, and establishing oxidative stress model and free radical model. Results showed that sulfated modification caused a significant change in chemical composition and the apparent structure of MP, but the degradation of the main chain of MP was inconspicuous. Sulfated Mesona chinensis Benth polysaccharide (SMP) exhibited excellent free radical scavenging activities and improved the protective effect of MP on cellular oxidative stress by increasing superoxide dismutase (SOD) activity and decreasing malondialdehyde (MDA) content. This study suggested that sulfated modification is an effective method to improve the antioxidant activities of MP, so as to expand the development and application of MP.

Catalytic co-pyrolysis of Alternanthera philoxeroides and peanut soapstock via a new continuous fast microwave pyrolysis system.

Continuous fast microwave catalytic co-pyrolysis of Alternanthera philoxeroides and peanut soapstock was studied using HZSM-5 as catalyst. The effects of catalyst temperature, feedstock-to-catalyst ratio, and A. philoxeroides-to-peanut soapstock ratio on the yield and composition of bio-oil were studied. Experimental results showed that the optimum catalyst temperature was 400 °C. The catalyst increased the proportion of aromatics but reduced the bio-oil yield. The optimum feedstock-to-catalyst ratio was 2:1. A. philoxeroides presented a significant synergistic effect with peanut soapstock, which facilitated the production of aromatics in the bio-oil. The optimum A. philoxeroides-to-peanut soapstock ratio was 1:2.

Conversion of woody oil into bio-oil in a downdraft reactor using a novel silicon carbide foam supported MCM41 composite catalyst.

This study reports the synthesis of a SiC-MCM41 composite catalyst by a microwave-assisted hydrothermal process and the composite catalyst had the characteristics of MCM41 and SiC, and the surface of SiC grew evenly with a layer of MCM41 after characterization of the catalysts by various means (X-ray diffraction, Brunauer-Emmett-Teller, scanning electron microscopy). The catalyst was applied in the pyrolysis of waste oil to investigate how it influences the bio-oil component proportion compared with no catalyst, only SiC, only MCM41 catalysis and the catalytic effect was also investigated at different temperatures and different catalyst to feed ratios. In a downdraft system with a pyrolysis temperature of 550 °C, a catalyst to feed ratio of 1 : 2, and a catalytic temperature of 400 °C, 32.43% C5-C12 hydrocarbons and 41.10% mono-aromatics were obtained. The composite catalyst combined the catalytic effect of SiC and MCM41 because it increased the amount of C5-C12 hydrocarbons and decreased the amount of oxygen-containing compounds in bio-oil. After repeated uses, the composite catalyst still retained the catalytic properties.

Production of bio-oil from agricultural waste by using a continuous fast microwave pyrolysis system.

In this study, a continuous fast microwave-assisted pyrolysis system was developed to produce bio-oil, gas, and biochar from rice straw and Camellia oleifera shell. The effects of different pyrolysis temperatures (400 °C, 500 °C, and 600 °C) and feed rates (rice straw: 25, 45, and 66 g/min; C. oleifera shell: 100, 200, and 400 g/min) on bio-oil production were investigated. Experimental results showed that the yields of bio-oil (31.86 wt%) and gas (54.49 wt%) produced by the microwave-assisted pyrolysis of rice straw increased with increasing temperature. By contrast, the yields of bio-oil (27.45 wt%) and biochar (35.47 wt%) produced by the pyrolysis of C. oleifera shell decreased with increasing temperature. The contents of phenols, aldehydes, and alcohols in bio-oil produced from the shell were higher than those in bio-oil derived from rice straw.

Anti-diabetic properties of Momordica charantia L. polysaccharide in alloxan-induced diabetic mice.

A water-soluble polysaccharide (MCP) was isolated from the fruits of Momordica charantia L., and the hypoglycemic effects of MCP were investigated in both normal healthy and alloxan-induced diabetic mice. MCP was orally administered once a day after 3 days of alloxan-induction at 100, 200 and 300mg/kg body weight for 28 day. Results showed that fasting blood glucose level (BGL) was significantly decreased, whereas the glucose tolerance was marked improvement in alloxan-induced diabetic mice, and loss in body weight was also prevented in diabetic mice compared to the diabetic control group. The dosage of 300mg/kg body weight exhibited the best effects. In addition, MCP did not exhibit any toxic symptoms in the limited toxicity evaluation in mice. The results suggest that MCP possess significantly dose-dependent anti-diabetic activity on alloxan-induced diabetic mice. Hence, MCP can be incorporated as a supplement in health-care food, drugs and/or combined with other hypoglycemic drugs.

[Determination of N-carbamyl-L-glutamic acid in feedstuff by high performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry].

A method was developed for the determination of N-carbamyl-L-glutamic acid (NCG) in feedstuff by high performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS). The feedstuff samples were extracted with methanol, cleaned-up by a ProElut mixed-mode of strong anion exchange inverse column (PXA). Then the samples were separated with HPLC and detected with MS/MS in multiple reaction monitoring (MRM) mode via positive electrospray ionization (ESI+). The fragment ions of m/z 148.0 and m/z 84.0 were used for qualitative analysis, and m/z 130.0 was selected for quantitative analysis. The limit of detection for NCG was 24 microg/kg (S/N > 3), and the limit of quantification was 80 microg/kg (S/N > 10). The standard calibration curve was linear over the range of 20-1,000 microg/L, and the correlation coefficient was 0. 999 9. The recoveries of NCG in feedstuff were 104.0%, 103.5%, 95.3%, at the three spiked levels of 80, 200, 500 mg/kg with the relative standard deviations (RSDs) of 7.5%, 6.3% and 5.8%. The results showed that the method is simple, fast, highly sensitive, specific and with good purification effect. It can be applied to the analysis and determination of NCG in feedstuff.