Characteristics of soy films as affected by transglutaminase cross-linking and inclusions of pectin and protein enhancers.

Previously, we showed that water extract (soymilk, except pH was increased to 8 from 6.5) of whole soybean could be used directly as a raw material for producing edible soy films by deposition of the film-forming solution (soy extract with enhancers). However, the strength of such soy films needed improvement because they were weak. The purpose of this study was to investigate how transglutaminase (TG) cross-linking reactions and film enhancers, including pectin (low- and high-methoxyl pectin), whey protein isolate (WPI), and soy protein isolate (SPI), improve the physical properties of soy films. Soy films prepared with TG had tensile strength (TS) of 3.01 MPa and puncture strength (PS) of 0.78 MPa, which were higher by as much as 51% and 30% than that of soy films without TG treatment, respectively. Pectin showed significant effects on the mechanical properties of TG-added soy films in terms of TS, PS, and % elongation. On the other hand, only TS and PS were increased by the addition of WPI or SPI. Heat curing had a significant effect on soy film's physical properties. TG treatment significantly reduced film solubility when soaked in water and various levels of acid (vinegar) and base (baking soda) solutions. Under the experimental conditions of 35 unit TG and 28 min of reaction, the degrees of cross-linking were evidenced by the disappearance of individual protein subunits, except the basic subunit of glycinin, and the reduction of 21% of lysine residues of the proteins. HIGHLIGHTS: Edible soy films were made with transglutaminase and about 21% lysine cross-linked. The mechanical strength of soy films was increased by incorporating film enhancers. Transglutaminase enhanced the mechanical properties of soy films.

A new method based on dispersive solid phase microextraction with commercial MOFs coupled to LC-MS/MS for the determination of isoflavones in soy drinks.

For the first time, a method based on dispersive solid phase microextraction (D-µSPE) using commercial metal-organic frameworks coupled to liquid chromatography-triple quadrupole tandem mass spectrometry (LC-MS/MS) has been proposed for the determination of isoflavones in soy drinks. The use of commercial sorbents simplifies the sample treatment procedure and allows their application to routine analysis. Optimization of the parameters involved in the microextraction process was carried out using a Box-Behnken experimental design. Under the optimized conditions, the limits of detection ranged between 2 and 7 µg L-1; the intra-day and inter-day precision were <10 and 20%, respectively, and the recoveries were in the range of 61-120%. No significant matrix effect was found, which allowed the use of external standard calibration method. The method was successfully applied to the determination of isoflavones in commercial soy milk samples.

Introducing Bacillus natto and Propionibacterium shermanii into soymilk fermentation: A promising strategy for quality improvement and bioactive peptide production during in vitro digestion.

Herein, the texture properties, polyphenol contents, and in vitro protein digestion characteristics of soymilk single- or co-fermented by non-typical milk fermenter Bacillus natto (B. natto), Propionibacterium freudenreichii subsp. shermanii (P. shermanii), and traditional milk fermenter were evaluated. Co-fermenting procedure containing B. natto or P. shermanii could raise the amounts of gallic acid, caffeic acid, and GABA when compared to the unfermented soymilk. Co-fermented soymilk has higher in vitro protein digestibility and nutritional protein quality. Through peptidomic analysis, the co-work of P. shermanii and Lactobacillus plantarum (L. plantarum) may release the highest relative percentage of bioactive peptides, while the intervention of B. natto and Streptococcus thermophilus (S. thermophilus) resulted in more differentiated peptides. The multi-functional bioactive peptides were mainly released from glycine-rich protein, ß-conglycinin alpha subunit 1, and ACB domain-containing protein. These findings indicated the potential usage of B. natto/S. thermophilus or P. shermanii/L. plantarum in bio-enhanced soymilk fermentation.

Novel strategy to raise the content of aglycone isoflavones in soymilk and gel: Effect of germination on the physicochemical properties.

Germination holds the key to nutritional equilibrium in plant grains. In this study, the effect of soybean germination on the processing of soymilk (SM) and glucono-δ-lactone (GDL) induced soymilk gel (SG) was investigated. Germination promoted soybean sprout (SS) growth by activating the energy metabolism system. The energy metabolism was high during the three-day germination and was the most vigorous on the second day of germination. After germination, protein dissolution was improved in SM, and endogenous enzymes produced small molecule proteins. Small molecule proteins were more likely to aggregate to produce SM protein particles. Germination increased the water-holding capacity of SG induced by GDL but weakened the strength. Furthermore, the dynamic fluctuations in isoflavone content were closely monitored throughout the processing of soybean products, including SS, SM, and SG. Although the total amount of isoflavones in SM and SG processed from germinated soybeans decreased, a significant enrichment in the content of aglycone isoflavones was observed. The content of aglycone isoflavones in SG processed from germinated soybeans on the second day of germination was 736.17 ± 28.49 µg/g DW, which was 83.19 % higher than that of the control group. This study demonstrates that germination can enhance the nutritional value of soybean products, providing innovative opportunities for the development of health-promoting soybean-based products.

Effect of preheating-induced denaturation of proteins and oleosomes on the structure of protein and soymilk properties.

In this paper, effects of preheating-induced denaturation of proteins and oleosomes on protein structure and soymilk quality were studied. The protein in soybeans baked at 55 °C (B-55) and 85 °C (B-85) showed an increase of ß-sheet content by 3.2 % and a decrease of α-helix content by 3.3 %, indicating that proteins were gradually unfolded while oleosomes remained intact. The protein resisted thermal denaturation during secondary heating, and soymilks were stable as reflected by a small d3,2 (0.4 µm). However, raw soymilk from soybeans baked at 115 °C (B-115), steamed for 1 min (ST-1) and 5 min (ST-5) presented oleosomes destruction and lipids aggregates. The proteins were coated around the oil aggregates. The ß-turn content from soybeans steamed for 10 min (ST-10) increased by 9.5 %, with a dense network where the OBs were tightly wrapped, indicating the serious protein denaturation. As a result, the soymilks B-115 or steamed ones were unstable as evidenced by the serious protein aggregation and larger d3,2 (5.65-12.48 µm). Furthermore, the soymilks were graininess and the protein digestion was delayed due to the formation of insoluble protein aggregates. The flavor and early-stage lipid digestion of soymilk from steamed soybeans was improved owing to lipid release.

A deep learning-based quantitative prediction model for the processing potentials of soybeans as soymilk raw materials.

Current technologies as correlation analysis, regression analysis and classification model, exhibited various limitations in the evaluation of soybean possessing potentials, including single, vague evaluation and failure of quantitative prediction, and thereby hindering more efficient and profitable soymilk industry. To solve this problem, 54 soybean cultivars and their corresponding soymilks were subjected to chemical, textural, and sensory analyses to obtain the soybean physicochemical nature (PN) and the soymilk profit and quality attribute (PQA) datasets. A deep-learning based model was established to quantitatively predict PQA data using PN data. Through 45 rounds of training with the stochastic gradient descent optimization, 9 remaining pairs of PN and PQA data were used for model validation. Results suggested that the overall prediction performance of the model showed significant improvements through iterative training, and the trained model eventually reached satisfying predictions (|relative error| ≤ 20%, standard deviation of relative error ≤ 40%) on 78% key soymilk PQAs. Future model training using big data may facilitate better prediction on soymilk odor qualities.

Analysis of Protein Digestion and Absorption Using a Ussing Chamber to Simulate the Environment in the Digestive Tract.

The Ussing chamber is a tool for analyzing drug absorption. We investigated whether the Ussing chamber can be used to analyze the process from digestion to absorption of protein in the gastrointestinal tract. Mixtures containing infant formula, whole cow's milk, processed soy milk, enteral nutrition, or human breast milk, were placed in the apical membrane side equipped with Caco-2 cells. After the addition of first pepsin then pancreatin, samples from the apical and basal membranes were collected. Infant formula showed the highest digestibility and absorption rate. This may be attributed to the presence of whey protein, which is rapidly digested and absorbed. The digestion and absorption of human breast milk showed different results in each donor, suggesting that digestion and absorption may vary among individuals. We concluded that the Ussing chamber can continuously analyze the process from digestion to absorption of proteins in the gastrointestinal tract.

Effects of water-soluble and water-insoluble α-glucans produced in situ by Leuconostoc citreum SH12 on physicochemical properties of fermented soymilk and their structural analysis.

This study investigated the effect of in situ produced water-soluble α-glucan (LcWSG) and water-insoluble α-glucan (LcWIG) from Leuconostoc citreum SH12 on the physicochemical properties of fermented soymilk. α-Glucans produced by Leuc. citreum SH12 improved water-holding capacity, viscosity, viscoelasticity and texture of fermented soymilk. Gtf1365 and Gtf836 of the five putative glucansucrases were responsible for synthesizing LcWSG and LcWIG during soymilk fermentation, respectively. Co-fermentation of soymilk with Gtf1365 and Gtf836 and non-exopolysaccharide-producing Lactiplantibacillus plantarum D1031 indicated that LcWSG effectively hindered the whey separation of fermented soymilk by increasing viscosity, while LcWIG improved hardness, springiness and accelerated protein coagulation. Fermented soymilk gel formation was mainly based on hydrogen bonding and hydrophobic interactions, which were promoted by both LcWSG and LcWIG. LcWIG has a greater effect on α-helix to ß-sheet translation in fermented soymilk, causing more rapid protein aggregation and thicker cross-linked gel network. Structure-based exploration of LcWSG and LcWIG from Leuc. citreum SH12 revealed their distinct roles in the physicochemical properties of fermented soymilk due to their different ratio of α-1,6 and α-1,3 glucosidic linkages and various side chain length. This study may guide the application of the water-soluble and water-insoluble α-glucans in fermented plant protein foods for their quality improvement.

Conformation-Activity Mechanism of Alcalase Hydrolysis for Reducing In Vitro Allergenicity of Instant Soy Milk Powder.

Effective reduction of the allergenicity of instant soy milk powder (ISMP) is practically valuable for expanding its applications. This study optimized the enzymolysis technology of ISMP using single-factor experiments and response surface methodology, combined serological analysis, cellular immunological models, bioinformatics tools, and multiple spectroscopy techniques to investigate the effects of alcalase hydrolysis on allergenicity, spatial conformation, and linear epitopes of ISMP. Under the optimal process, special IgE and IgG1 binding abilities and allergenic activity to induce cell degranulation of alcalase-hydrolyzed ISMP were reduced by (64.72 ± 1.76)%, (56.79 ± 3.72)%, and (73.3 ± 1.19)%, respectively (P < 0.05). Moreover, the spatial conformation of instant soy milk powder hydrolysates (ISMPH) changed, including decreased surface hydrophobicity, a weaker peak of amide II band, lower contents of α-helix and ß-sheet, and an enhanced content of random coil. Furthermore, the linear epitopes of major soy allergens, 9 from glycinin and 13 from ß-conglycinin, could be directionally disrupted by alcalase hydrolysis. Overall, the structure-activity mechanism of alcalase hydrolysis to reduce ISMP allergenicity in vitro was preliminarily clarified. It provided a new research direction for the breakthrough in the desensitization of ISMP and a theoretical basis for revealing the potential mechanism of alcalase enzymolysis to reduce the allergenicity of ISMP.

Effect and mechanism of freezing on the quality and structure of soymilk gel induced by different salt ions.

BACKGROUND: The increasing attention toward frozen soy-based foods has sparked interest. Variations exist in the quality and structure of soymilk gels induced by different salt ions, leading to diverse changes post-freezing. This study compared and analyzed the effects of calcium chloride (CC), magnesium chloride (MC) and calcium sulfate (CS) on the quality characteristics and protein structure changes of soymilk gels (CC-S, MC-S and CS-S) before and after freezing, and clarified the mechanisms of freezing on soymilk gel. RESULTS: The formation rate of soymilk gel is influenced by the type of salt ions. In comparison to CS and MC, soymilk gel induced by CC exhibited the fastest formation rate, highest gel hardness, lowest moisture content, and smaller gel pores. However, freezing treatment deteriorated the quality of soymilk gel induced by different salt ions, leading to a decline in textural properties (hardness and chewiness). Among these, the textual state of CC-induced soymilk gel remained optimal, exhibiting the least apparent damage and minimal cooking loss. Freezing treatments prompt a transition of soymilk gel secondary structure from ß-turns to ß-sheets, disrupting the protein's tertiary structure. Furthermore, freezing treatments also fostered the crosslinking between soymilk gel protein, increasing the content of disulfide bonds. CONCLUSION: The quality of frozen soymilk gel is influenced by the rate of gel formation induced by salt ions. After freezing, soymilk gel with faster gelation rates exhibited a greater tendency for the transformation of protein-water interactions into protein-protein interactions. They showed a higher degree of disulfide bond formation, resulting in a more tightly knit and firm frozen gel network structure with denser and more uniformly distributed pores. © 2024 Society of Chemical Industry.

Effects of removing phytic acid on the bioaccessibility of Ca/Fe/Zn and protein digestion in soymilk.

BACKGROUND: Soymilk is a high-quality source of protein and minerals, such as calcium (Ca), iron (Fe), and zinc (Zn). However, phytic acid in soymilk restricts mineral and protein availability. We here investigated the effects of removing phytic acid on the physicochemical properties, mineral (Ca, Fe, and Zn) bioaccessibility, and protein digestibility of soymilk. RESULTS: Physicochemical property analysis revealed that the removal of phytic acid reduced protein accumulation at the gastric stage, thereby facilitating soymilk matrix digestion. The removal of phytic acid significantly increased Zn bioaccessibility by 18.19% in low-protein soymilk and Ca and Fe bioaccessibility by 31.20% and 30.03%, respectively, in high-protein soymilk. CONCLUSION: Removing phytic acid was beneficial for the hydrolysis of high-molecular-weight proteins and increased the soluble protein content in soymilk, which was conducive to protein digestion. This study offers a feasible guide for developing plant-based milk with high nutrient bioaccessibility. © 2024 Society of Chemical Industry.

Enhancing the functionality of plant-based Yogurt: Integration of lycopene through dual-stage fermentation of soymilk.

Well-defined compositional assemblies of plant-based yogurt are of fast-growing awareness for world population concerning environmental sustainability, economic burdens and health risks. Soybean is an attractive candidate for plant yogurt, suffering from poor flavor, limited nutrition, and undesired allergens to offer healthy-functional segments. Herein, we deciphered a novel lycopene-soy yogurt by efficient two-stage fermentation of engineered B. subtilis and LAB. The fortified sogurt was ensured with redundant lycopene of 22.67 ± 2.95 mg/g DCW by engineered B. subtilis and enriched soy isoflavone from synergistic effects of engineered B. subtilis and LAB, possessing strong antioxidant capacity for upgrading functionality. Moreover, the desired pH, accelerated protein hydrolysis, enhanced amino acid availability, and expected sensory attributes cooperatively conferred lycopene-soy yogurt as healthy functional food. High potential is firstly ascribed to sequential dual culture of engineered B. subtilis and LAB in lycopene-soy yogurt, in which flavorful, hypoallergenic and antioxidative ingredients enabled functionalities for plant-based yogurt.

Comparison between hydrodynamic and ultrasound cavitation on the inactivation of lipoxygenase and physicochemical properties of soy milk.

The effects of hydrodynamic cavitation (HC) and ultrasound cavitation (UC) on the lipoxygenase activity and physicochemical properties of soy milk were evaluated. The results revealed that both ultrasound cavitation and hydrodynamic cavitation significantly inactivated the lipoxygenase activity. After the exposure to ultrasound cavitation at 522.5 W/L and 70 °C for 12 min, the lipoxygenase activity was inactivated by 96.47 %. Meanwhile, HC treatment with the cavitation number of 0.0133 for 240 min led to the loss of 79.31 % of lipoxygenase activity. An artificial neural network was used to model and visualize the effects of different parameters after ultrasound cavitation treatment on the inactivation efficiency of soy milk. Turbiscan test results showed that hydrodynamic and ultrasound cavitation decreased the instability index and particle size of soy milk. Moreover, the total free amino acid content was significantly increased after hydrodynamic and ultrasound cavitation treatment. Gas chromatography-mass spectrometry showed that the total content of beany flavor compounds decreased after acoustic cavitation and HC treatment. Acoustic cavitation and HC affected the tertiary and secondary structure of soy milk, which was related to the inactivation of lipoxygenase. We aim to explore a potential and effective way of the application in soy milk processing by comparing the ultrasound equipped with heat treatment and hydrodymic cavitation.

An Investigation into the Performance and Mechanisms of Soymilk-Sized Handmade Xuan Paper at Different Concentrations of Soymilk.

Invaluable paper relics that embody a rich traditional culture have suffered damage, requiring urgent restoration. In this context, the utilization of soymilk as a sizing agent holds great significance and reverence. This study investigates the use of soymilk as a sizing agent for Xuan paper and evaluates its effects on various properties and the long-term behavior of the paper. The findings reveal that the application of soymilk as a sizing agent for Xuan paper imparts distinct properties, including hydrophobicity, improved mechanical properties, and unique chromaticity. These characteristics-arising from the papillae on the surface of the Xuan paper, the protein folding of the soy protein, and hydrogen-bonding interactions between the soy protein and paper fibers-play a crucial role in shaping the paper's unique attributes. From a physicochemical perspective, the aging process leads to multiple changes in paper properties. These changes include acidification, which refers to a decrease in pH, as well as a decline in mechanical strength, an increase in chromaticity, and a decrease in the degree of polymerization (DP) of the paper. The Ekenstam equation is employed to predict the lifespan of the paper, showing longer lifespans for Sheng Xuan paper and a negative correlation between soymilk concentration and lifespan in soymilk-sized paper. Our work provides valuable insights for the preservation and maintenance of paper, highlighting the potential benefits and challenges of using soymilk for surface sizing.

Evaluation of nutritional and physicochemical characteristics of soy yogurt by Lactobacillus plantarum KU985432 and Saccharomyces boulardii CNCMI-745.

Nutritional yeast-produced soy yogurt has grown in demand, because of its unique nutritional and health benefits. It has low cholesterol, no lactose, and high levels of protein, probiotic yeast, vitamins, and minerals. In this work, Soymilk (12.5%) was prepared and fermented to produce soy yogurt. Growth curves, probiotic characteristics of Saccharomyces boulardii CNCMI-745 and Lactobacillus plantarum KU985432 were determined. The nutritional value of both yogurts was evaluated, including viable cell count, protein, vitamin B-complex, sugars, phenolic acids, and fatty acids, mineral content, stability, and storage. Analysis of the physicochemical composition of the yogurts included assessment of titratable acidity, antioxidant potential, viscosity, and moisture content. The probiotic viable count of the produced yogurts met the standards for commercial yogurts. S. boulardii CNCMI-745 displayed safety characteristics and high tolerance to heat, acid, and alkaline stress. The produced B vitamins increased in both yogurts. The total saturated fatty acids in Saccharomyces-yogurt decreased, while the unsaturated fatty acids increased. Saccharomyces-yogurt showed high antioxidant activity, phenolic acids, and crude protein content. Both yogurts demonstrated the same tendency for stability during 16 day-storage. In conclusion, using nutritional yeast in the production of soy yogurt increased its nutritional content more than probiotic lactic acid bacteria.

Nutritional and functional insight into novel probiotic lycopene-soy milk by genome edited Bacillus subtilis.

Nutritional and functional soy-based milk gains growing attention globally in food industry. However, its poor sensorial attributes, single flavor, and limited substance variety become critical issues in displaying balanced nutrition and multifunction for health. Herein, a novel probiotic lycopene-soy milk was developed by genome edited Bacillus subtilis harboring lycopene biosynthesis cassette with efficient lycopene production of 25.73 ±â€¯1.57 mg/g DCW. Further investigation displayed desirable pH, reducing sugar, protein, total phenolic content and isoflavone for achieved milk than conventional soy milk, implying it with well-balanced nutritional quality. Notably, achieved milk exhibited stronger antioxidant capacity and higher isoflavone bioavailability for functionality. Moreover, it possessed significantly high scores for taste, appearance, and overall acceptability, suggesting its excellent sensorial attributes. To our delight, it is the first time to fortify soy-milk with probiotic and lycopene by genome edited B. subtilis to explore additive effect on improving nutritional value and functionality for food application.

Effect of cavitation jet technology on instant solubility characteristics of soymilk flour: Based on the change of protein conformation in soymilk.

The protein conformation of soymilk is the key to affecting the instant solubility of soymilk flour. This study aimed to evaluate the effect of cavitation jet treatment time (0, 2, 4, 6, and 8 min) on the instant solubility of soymilk flour based on the conformational changes of protein in soymilk. The results showed that the cavitation jet treatment for 0-4 min significantly unfolded the protein structure of soymilk and increased the content of soluble protein, which reduced the particle size and increased the electrostaticrepulsion and the viscosity of soymilk. This was beneficial for soymilk droplets fully atomized and repolymerized in the spray drying tower, forming soymilk flour particles with large size, smooth surface, and uniform distribution. When the cavitation jet treatment time was 4 min, the wettability (from 127.3 ± 2.5 s to 84.7 ± 2.1 s), dispersibility (from 70.0 ± 2.0 s to 55.7 ± 2.1 s), and solubility (from 56.54% to 78.10%) of soymilk flour were significantly improved. However, when the time of the cavitation jet treatment was extended to 8 min, the protein of soymilk aggregated and the stability of soymilk decreased, which reduced the particle size and hurt the surfacecharacteristics of soymilk flour after spraydrying. It resulted in a decrease in the instant solubility of soymilk flour. Therefore, the cavitationjet treatment with proper time increases the instant solubility of soymilk flour by improving the protein conformation of soymilk.

Effect of a co-fermentation system with high-GABA-yielding strains on soymilk properties: microbiological, physicochemical, and aromatic characterisations.

Biosynthesis is the safest method for preparing GABA; however, there are not enough GABA-producing strains to provide an effective resource. The purpose of this study was to determine the feasibility of using Lactobacillus fermentum SMN10-3(A) and Lactococcus lactis SMN15-6(B) to study the effects of strain complex pairing on the GABA formation, flavour, and metabolic pathways of fermented soymilk. It was found that group A2B1 had the highest acid production rate, GABA yield (1.76 ± 0.01 mg/mL), and flavour compound content. A total of 55 differential metabolites were produced after fermentation, of which 28 dominated by hexanal were significantly downregulated and 26 dominated by alcohols were significantly upregulated. The significant metabolic pathways involved were d-alanine, taurine and hypotaurine, and selenocompound metabolism. Finally, the components contributing to the aroma of fermented soymilk were identified, which included 2-pentylfuran and 2-butyl-2-octenal. These results provide a theoretical basis for future research on GABA-rich fermented foods.

Mechanism of the glucono-δ-lactone induced soymilk gelation: Enthalpy and entropy transformation in the cross-linking of protein molecules.

This study aimed to explore new techniques to regulate the quality of soy products. The glucono-δ-lactone (GDL) induced soymilk gelation process and the gel network structure characteristic were compared as a matter of temperature, and the role of reaction kinetics was discussed. Results showed that there were similarities in the development of G' curves under different temperatures, whereas the gel network structures and the energy requirements of cross-linking reactions were different. In the high-temperature region (70 °C-95 °C), the exposure and binding of reactive groups were promoted. The activation enthalpy (ΔH*) required by protein aggregates decreased and the effect of entropy reduction (-TΔS*) was enhanced, which led to shorten the preaggregation time (tg) and increase the gelation rate (k), resulting in the formation of rough, porous gel network with high stiffness. By contrast, in the low-temperature region (40 °C-70 °C), high enthalpy contributions and low entropy changes were required, then a fine, soft, and tender gel network formed. Besides, a funnel-shaped model of the enthalpy-entropy energy transformation mechanism of soymilk gelation was proposed. The results of this study revealed that adjusting the enthalpy-entropy energy requirements of the protein cross-linking reaction could be utilized to the regulation of the network structure and quality of soymilk gels, which could enrich the reaction kinetics theory and provide innovative ideas for food quality control technology from the perspective of energy requirement and energy input.

The fortification of encapsulated soy isoflavones and texture modification of soy milk by α-lactalbumin nanotubes.

Nanocarriers can improve the dispersibility of hydrophobic bioactive compounds and potentially improve the texture of liquid food formulations. Here, nanotubes (NTs) with a high aspect ratio formed by self-assembly of peptides partially hydrolyzed from α-lactalbumin (α-lac) were used to deliver soy isoflavones (IFs) and modify soy milk texture. IFs encapsulated by nanotube (NT/IFs) via hydrophobic interactions, which had improved dispersibility, with a maximum loading efficiency of 4%. The rheological characterization showed that the nanotubes enhanced the viscoelastic property and long term-stability of soy milk. About 80% of the NT/IFs in soy milk survived simulated in in vitro gastric digestion promoting the release of IFs in the intestinal phase. Overall, this work demonstrated that α-lac nanotubes may be a multi-functional carrier system for hydrophobic compounds providing beneficial changes to functional food texture.