In this study, a variety of probiotic strains, including Lactiplantibacillus plantarum, Lacticaseibacillus casei, Lactobacillus acidophilus, Streptococcus thermophilus, Bifidobacterium longum, Limosilactobacillus reuteri, Lactobacillus delbrueckii subsp. bulgaricus, Lacticaseibacillus rhamnosus, and Bifidobacterium bifidum, were utilized for soymilk fermentation both as free cells and as synbiotics on agro-industrial residuals such as okara, whey protein, banana peels, apple pomace, sugarcane bagasse, orange peels, and lemon peels. Among these, Lacticaseibacillus rhamnosus emerged as the most significant strain for soymilk fermentation, exhibiting a viability of 10.47 log cfu/mL, a pH of 4.41, total acidity of 1.12%, and organic acid contents (lactic and acetic acid) of 11.20 and 7.50 g/L, respectively. As a synbiotic Lacticaseibacillus rhamnosus immobilised on okara, showed even more impressive results, with a viability of 12.98 log cfu/mL, a pH of 4.31, total acidity of 1.27%, and organic acid contents of 13.90 and 9.30 g/L, respectively. Over a 12-h fermentation period, cell viability values increased by 10.47-fold in free cells and 11.19-fold in synbiotics. Synbiotic supplementation of fermented soymilk proved more beneficial than free cells in terms of viability, acidity, and organic acid content. Furthermore, when synbiotic fermented soymilk was freeze-dried to simulate the digestive system in vitro, synbiotics and freeze-dried cells demonstrated superior gastrointestinal tract survival compared to free cells. Both the probiotic bacteria and the synbiotics exhibited cytotoxicity against colon and liver cancer cell lines, with half-maximal inhibitory concentrations ranging from 41.96 to 61.52 μL/well.

LCPS-1, a cell wall polysaccharide (CWPS), is bound to the cell wall of the probiotic Lacticaseibacillus paracasei (formerly known as Lactobacillus casei) strain Shirota (LcS). Generally, the role of CWPS in the viability and survivability of bacteria is yet to be fully understood. This study aimed to elucidate the role of LCPS-1 in the viability and survivability of LcS. A mutant strain completely lacking LCPS-1 was constructed and evaluated for growth in bovine and soy milk and susceptibility to acid and bile. The growth of the mutant in bovine and soy milk temporarily stalled after the late logarithmic phase while wild-type LcS continued growing, resulting in a significantly lower number of viable cells for the mutant strain (p < 0.01). Significantly higher cell death relative to that of the wild-type strain was observed for the mutant strain following acid treatment at pH 3.0 (p < 0.01), with 60 and 92 % survival, respectively. The absence of LCPS-1 also reduced the survival rate of LcS cells from 3.3 to 0.8 % following 0.2 % bile treatment. The survival rate of the mutant after consecutive treatment with acid and bile was 19 %, while 73 % of the wild-type LcS survived. These results indicate that LCPS-1 leads to higher LcS growth in milk and improves tolerance to acid and bile. This study reveals the contribution of probiotic bacterial CWPS to acidic and gastrointestinal stress tolerance. Based on these findings, characterizing and modifying CWPS in probiotic strains could enhance manufacturing yields and improve gastrointestinal stress tolerance after consumption by hosts, ultimately advancing the development of more effective probiotics.

The mixed yogurt was fermented from Cow-Soy milk and modified by transglutaminase (TG). The effects of mixed milk and TG on the quality characteristics of mixed yogurt were investigated by texture characteristics, rheology (rheometer) and structure (scanning electron microscopy). The findings revealed that the mixed yogurt with 50% cow milk exhibited lower hardness, viscosity and consistency. Furthermore, when TG was added, the yogurt showed better rheological properties, sensory score and a more stable microstructure. Compared with the samples without TG modification, the viscosity and cohesiveness of the modified samples increased by 10% and 100%, respectively. The combination of cow milk and soy milk improved the texture of yogurt, and the TG addition further improved the physicochemical properties of yogurt. This finding provided a meaningful reference for the development of mixed yogurt with a suitable taste from animal and plant milk, and laid a basis for the practical application of mixed yogurt in the dairy industry, which will meet the requirements for dairy products for consumers in future.

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.

BACKGROUND: Resistance exercise training (RET) can increase muscle mass and strength, and this adaptation is optimized when dietary protein is consumed to enhance muscle protein synthesis. Dairy milk has been endorsed for this purpose; however, allergy and lactose intolerance affect two-thirds of the global population making dairy milk unsuitable for many. Plant-based alternatives such as soy milk have gained popularity and exhibit comparable protein content. However, concerns regarding soy phytoestrogens potentially influencing circulating sex hormones and diminishing the anabolic response to RET have been raised. This study therefore aimed to assess the acute effects of dairy and soy milk consumption on circulating sex hormones (total, free testosterone, free testosterone percentage, total estrogen, progesterone, and sex hormone binding globulin) after RET.
METHODS: Six male participants were recruited for a double-blinded, randomized crossover study with either dairy or soy milk provided post RET. Venous samples were collected before and after milk consumption across seven timepoints (0-120 minutes) where circulating sex hormones were analyzed. Two-way ANOVA analyses were applied for repeated measures for each hormone. The area under the curve (AUC) was also calculated between dairy and soy milk. Significance was set at p<0.05.
RESULTS: No significant differences were observed in acute circulating serum for free (p=0.95), % free (p=0.56), and total testosterone (p=0.88), progesterone (p=0.67), or estrogen (p=0.21) between milk conditions. Likewise, no significant differences in AUC were observed between any hormones.
CONCLUSIONS: These findings suggest that consumption of dairy milk and soy milk have comparable acute effects on circulating sex hormones following RET. Further investigations with expanded sample sizes are needed to strengthen and broaden these initial findings.

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, β-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.

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.

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.

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.

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 β-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 β-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.