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.

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.

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.

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 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.

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.

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.

The Effects of Dairy and Plant-Based Liquid Components on Lutein Liberation in Spinach Smoothies.

Lutein is a dietary lipophilic compound with anti-inflammatory properties. We have previously shown that dairy fat can improve the lutein content in spinach smoothies. It is, however, unclear whether fat concentrations and fermentation status in dairy products affect lutein liberation in smoothies. Moreover, plant-based milks vary in fat, protein, and fiber content which may affect lutein dissolution. This study aimed to provide translatable information to consumers by comparing lutein liberation in spinach smoothies made with different dairy or plant-based liquids in domestic settings. The smoothies were digested in vitro, and liberated lutein was measured by high-performance liquid chromatography (HPLC). High-fat and medium-fat cow's milk, as well as coconut milk with and without additives, were found to significantly improve lutein liberation by 36%, 30%, 25%, and 42%, respectively, compared to blending spinach with water alone. Adjustment models suggested that the effects of cow's milk and coconut milk were derived from fat and protein, respectively. On the other hand, soymilk with and without additives showed significantly reduced lutein liberation by 40% and 61%, respectively. To summarize, only 4 out of 14 tested liquids increased lutein liberation in spinach smoothies. The results highlight the importance of testing food companions for lipophilic active ingredients.

Viscosity of evaporated soymilk prepared in the laboratory using normal and 11S-lacking soybean seeds.

BACKGROUND: Soymilk is utilized not only as a beverage but also as an alternative to bovine milk, including products such as yoghurt and cream. Evaporated soymilk is expected to be utilized as condensed milk. Raw and heated soymilk samples prepared in our laboratory were evaporated and then subjected to viscosity measurement. The soymilk samples were made from two different varieties: Fukuyutaka, which contains 7S and 11S globulin proteins; and an 11S-lacking soybean (Nanahomare). RESULTS: Raw Fukuyutaka soymilk had a lower viscosity and could be concentrated to a solids content of over 300 g kg-1 compared to heated soymilk (around 250 g kg-1 ), but the viscosity changes of Nanahomare soymilk showed an opposite trend. Only 7S globulin was denatured during evaporation at 75 °C and likely affected the interaction between proteins and oil bodies. This tendency was remarkable in the Nanahomare soymilk. The strange viscosity change behavior of evaporated Nanahomare soymilk, number of protein particles, intrinsic fluorescence and flow behavior suggest that thermally denatured 7S globulin accelerates the interactions between oil bodies, whereas 11S globulin, which is probably in its native state, suppresses the acceleration by denatured 7S globulin. CONCLUSION: Raw soymilk containing native globulins shows a slower increase in viscosity during evaporation. However, denatured 7S globulin accelerates the increase in viscosity during evaporation through interactions between oil bodies. The effect of the denatured state of individual proteins on interactions is expected to be useful in understanding the interaction between proteins and in controlling their properties and functions. © 2022 Society of Chemical Industry.

Gelation properties of tofu induced by different coagulants: Effects of molecular interactions between nano-sized okara dietary fiber and soybean proteins.

Gelation of soybean proteins with different coagulates as affected by okara nano-sized dietary fiber (NDF) were investigated. As compared with the MgCl2 tofu, the tofu induced by glocono-δ-lactone was softer with higher moisture and denser microstructure. As the volumetric ratio of NDF increased, pH, viscosity, particle size, and zeta potential of the NDF soymilk steadily increased. Regardless of coagulate type, tofu proteins' prevalence of α-helix + ß-turn increased with the NDF ratio up to 40 % while ß-sheet + random coil decreased. At the same time, intermolecular interactions (predominate hydrophobic interaction) significantly decreased, leading to form more porous structures in the two types of tofu. Accordingly, yield and moisture of tofu increased gradually with the NDF ratio, water holding capacity increased and then decreased, hardness decreased continuously. Results suggested that NDF affected the secondary structure and intermolecular interactions of soymilk proteins, which was independent of coagulate.

Comparison on Protein Bioaccessibility of Soymilk Gels Induced by Glucono-δ-Lactone and Lactic Acid Bacteria.

In this study, the protein bioaccessibility of soymilk gels produced by the addition of glu-cono-δ-lactone (GDL) and fermentation with lactic acid bacteria (LAB) was examined using an in vitro gastrointestinal simulated digestion model. The in vitro protein digestibility, soluble protein content, free amino acids contents, degree of hydrolysis, electrophoretic patterns, and peptide content were measured. The results suggested that acid-induced soymilk gel generated by GDL (SG) showed considerably reduced in vitro protein digestibility of 75.33 ± 1.00% compared to the soymilk gel induced by LAB (SL) of 80.57 ± 1.53% (p < 0.05). During the gastric digestion stage, dramatically higher (p < 0.05) soluble protein contents were observed in the SG (4.79−5.05 mg/mL) than that of SL (4.31−4.35 mg/mL). However, during the later intestinal digestion phase, the results were the opposite. At the end of the gastrointestinal digestion phase, the content of small peptides was not significantly different (p > 0.05) between the SL (2.15 ± 0.03 mg/mL) and SG (2.17 ± 0.01 mg/mL), but SL showed higher content of free amino acids (20.637 g/L) than that of SG (19.851 g/L). In general, soymilk gel induced by LAB had a higher protein bioaccessibility than the soymilk gel coagulated by GDL.

Engineering of Leuconostoc citreum for Efficient Bioconversion of Soy Isoflavone Glycosides to Their Aglycone Forms.

Soy isoflavones are phytochemicals that possess various beneficial physiological properties such as anti-aging, anti-tumor, and antioxidant properties. Since soy isoflavones exist in glycoside forms, their bioavailability requires initial hydrolysis of the sugar moieties bound to them to be efficiently absorbed through the gut epithelium. Instead of conventional chemical hydrolysis using acids or organic solvents, alternative strategies for enhancing the bioavailability of soy isoflavones using biological methods are gaining attention. Here, we engineered Leuconostoc citreum isolated from Korean kimchi for efficient bioconversion of soy isoflavone glycosides into their aglycone forms to enhance their bioavailability. We first constructed an expression module based on the isoflavone hydrolase (IH)-encoding gene of Bifidobacterium lactis, which mediates conversion of isoflavone glycosides to aglycone forms. Using a high copy number plasmid and bicistronic expression design, the IH was successfully synthesized in L. citreum. Additionally, we determined enzymatic activity of the IH using an in vivo ß-glucosidase assay and confirmed its highly efficient bioconversion efficiency for various types of isoflavone glycosides. Finally, we successfully demonstrated that the engineered L. citreum could convert isoflavone glycosides present in fermented soymilk into aglycones.

Comparative nutritional and antimicrobial analysis of Himalayan black and yellow soybean and their okara.

BACKGROUND: Soybean is believed to have good nutraceutical potential which is important for human health. Yellow soybean (YS) is generally used for the production of soymilk and other products, while black soybean (BS) is less explored. During the production of soymilk, residue, called okara is generated which is reported to have a good amount of nutrient content. Studies are generally performed with YS while BS is less explored. The present work is a comparison of the nutraceutical potential of BS and YS and their okara, mainly in terms of proximate, minerals, antinutrients, and isoflavone content and bioactivity of all types of samples in terms of antioxidant and antimicrobial activity. RESULTS: Compared to raw soybean, protein content decreased significantly in both types of okara. Phytochemicals like ascorbic acid, catechin, quercetin, and gallic acid were significantly (P < 0.05) high in BS residue in comparison to respective raw soybean. Among isoflavones, daidzin and genistin were found significantly varying among all the samples, and glycitin and glycitein were not present in YS. CONCLUSION: The nutraceutical potential and antimicrobial activity were comparative for both the raw beans and their okara, while the phytochemical contents and antioxidant activity were higher in the case of BS and its okara. © 2022 Society of Chemical Industry.

Effect of ultrasound on functional properties, flavor characteristics, and storage stability of soybean milk.

The effects of different ultrasound treatments (20 kHz at 400 W for 0 to 9 min) on the functional properties, flavor characteristics, and storage stability of soybean milk at 4 °C were investigated. Results indicated that non-sonicated soymilk had the maximum particle size D4, 3 of 2.47 ± 0.47 µm, while 9 min high intensity ultrasound (HIU) decreased D4, 3 to 0.44 ± 0.01 µm. 9 min of HIU decreased the total number of microorganisms in soymilk from 4.51 to 3.95 Log (CFU/mL). Moreover, 9 min HIU increased the absolute value of ζ-potential from 36.43 to 34.13 mV. Turbiscan test showed that 9 min HIU decreased the instability index of soymilk from 0.78 to 0.65. Furthermore, sensory analysis, electronic nose, electronic tongue, and gas chromatography-mass spectrometry showed that 7 min HIU decreased the content of aldehydes, furans, ketones, and alcohols by 52.09%, 75.01%, 56.79%, and 57.27%, respectively.