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.

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.

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.

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.

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.

Promising probiotic-fermented soymilk for alleviating acute diarrhea: insights into the microbiome and metabolomics.

Fermented soymilk (FSM4) has attracted much attention due to its nutritional and health characteristics. Exploring FSM4 products to alleviate diarrhea can ensure their effectiveness as a therapeutic food for alleviating gastrointestinal disorders. However, the relationship between gut microbiota and gut metabolite production remains unknown during diarrheal episodes. Therefore, the diarrhea-alleviating role and mechanisms of FSM4 in diarrhea rats were investigated via biochemical, gut microbiota, and serum metabolite analyses. The findings showed that consuming FSM4 improved diarrhea symptoms and reduced systemic inflammation better than non-fermented soymilk (NFSM). It is worth noting that FSM4 promoted the diversity, richness, structure, and composition of gut microbiota. It increased the ability to reduce inflammation associated with harmful bacteria (Anaerofilum, Flavonifractor, Bilophila, Anaerostipes, [Ruminococcus]_torques_group, Clostridium_sensu_stricto_1, Turicibacter, Ruminococcus_1, Ruminiclostridium_6, Prevotellaceae_NK3B31_group and Fusicatenibacter), while stimulating the growth of healthy species (Lactobacillus, Ruminococcaceae_UCG-014, Oscillibacter, [Eubacterium]_coprostanoligenes_group, Negativibacillus, and Erysipelotrichaceae_UCG-003). Moreover, metabolomics analysis showed that lipid metabolites such as lysophosphatidylethanolamine (LysoPE) and sphingolipids were upregulated in the NG group, closely related to pro-inflammatory cytokines (IL-6, IL-1ß, TNF-α, and IFN-γ) and the aforementioned pathogenic bacteria. Notably, in treatment groups, especially FSM4, the accumulation of L-ornithine, aspartic acid, ursocholic acid, 18-oxooleate, and cyclopentanethiol was increased, which was robustly associated with the anti-inflammatory factor IL-10 and beneficial bacteria mentioned above. Therefore, it can be inferred that the amino acids, bile acid, 18-oxooleate, and cyclopentanethiol produced in the FSM4 group can serve as metabolic biomarkers, which synergistically act with the gut microbiota to help alleviate inflammation for diarrhea remission. Overall, FSM4 may provide a new alternative, as an anti-inflammatory diet, to alleviate diarrhea.

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.

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 fermented soymilk with Lacticaseibacillus paracasei YIT 9029 on gut microbiota and defecation habits: a randomised, double-blind, placebo-controlled study.

Previous studies have demonstrated that soymilk and Lacticaseibacillus paracasei YIT 9029 (strain Shirota: LcS) each beneficially affect the gut microbiota and defecation habits. To investigate the effects of daily consumption of fermented soymilk containing LcS (FSM), we conducted a randomised, double-blind, placebo-controlled study of 112 healthy Japanese adults with a low faecal Bifidobacterium count. They consumed 100 ml FSM or placebo (unfermented soymilk base) once daily for 4 weeks. Their gut microbiota was analysed by 16S rRNA gene amplicon sequencing and quantitative reverse transcription-polymerase chain reaction (PCR), and faecal short-chain fatty acids (SCFAs) and urinary putrefactive products were assessed during the pre- and post-consumption periods. Defecation habits were examined weekly using a subjective questionnaire. In the post-consumption period, living LcS were not detected in two subjects in the FSM group (n = 57) but were detected in one subject in the SM group (n = 55). The FSM group had a significantly higher number and relative abundance of faecal lactobacilli compared with the placebo group. The relative abundance of Bifidobacterium, alpha-diversity of microbiota, and concentrations of acetate and total SCFAs in faeces were significantly increased in the FSM group, although no significant differences were detected between the groups. The number of defecations and defecation days per week significantly increased in both groups. Subgroup analysis of 109 subjects, excluding 3 with inconsistent LcS detection (2 and 1 subjects in the FSM and SM groups, respectively), revealed that the FSM group (n = 55) had significantly greater increases in faecal acetate concentration compared with the SM group (n = 54) and significant upregulation of pathways related to energy production or glucose metabolism in the gut microbiota. These findings suggest that daily FSM consumption improves the gut microbiota and intestinal environment in healthy adults and may help to maintain health and prevent diseases. Registered at the University Hospital Medical Information Network (UMIN) clinical trials registry under: UMIN 000035612.

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.

Breeding exceptionally fragrant soybeans for soy milk with strong aroma.

Knockout of the soybean (Glycine max) betaine aldehyde dehydrogenase genes GmBADH1 and GmBADH2 using CRISPR/Cas12i3 enhances the aroma of soybeans. Soy milk made from the gmbadh1/2 double mutant seeds exhibits a much stronger aroma, which consumers prefer; this mutant has potential for enhancing quality in soy-based products.

Effects of pH and salt concentration on freeze-thaw fractionation of soymilk protein.

BACKGROUND: The two major storage proteins of soymilk are the globulins 7S and 11S. Freeze-thaw fractionation is a simple method for separating these proteins in raw soymilk. In this study, we assessed the freeze-thaw fractionation ability of raw soymilk under various pH (4.3-11.6) conditions and added salt (sodium chloride) concentrations (0.00-0.67 mol L-1). RESULTS: We successfully achieved fractionation within a pH range of 5.8-6.7 and when the salt concentration was 0.22 mol L-1 or lower. Analysis of particle size distribution and microscopic examination of soymilk revealed no direct correlation between particle size and freeze-thaw fractionation ability. Interestingly, it was confirmed that the ranges of zeta potential values associated with successful freeze-thaw fractionation in raw soymilk remained consistent across different pH and salt concentration conditions. These ranges were between -23 and -28 mV at pH levels ranging from 5.8 to 6.7 and between -18 and -29 mV at added salt concentrations ranging from 0 to 0.22 mol L-1. CONCLUSION: The pH and salt concentration in raw soymilk markedly influence the freeze-thaw fractionation process. We confirmed that the range of zeta potential values where fractionation was possible remained consistent under various pH and salt concentration conditions. These findings suggest that the zeta potential value might serve as an indicator for evaluating the freeze-thaw fractionation ability of raw soymilk. © 2024 Society of Chemical Industry.

Probiotic Lactiplantibacillus plantarum KU210152 and its fermented soy milk attenuates oxidative stress in neuroblastoma cells.

We evaluated the probiotic properties and neuroprotective effects of Lactiplantibacillus plantarum KU210152 and its application in soy milk. L. plantarum KU210152 exhibited high tolerance to artificial gastrointestinal conditions, high adhesion to intestinal cells (HT-29), and safe enzyme production. Conditioned medium acquired from HT-29 cells treated with heat-killed lactic acid bacteria (LAB-CM) was used to evaluate the neuroprotective effects. The CM exhibited neuroprotective effects via cell viability assay, morphological observations, and suppression of ROS production. Heat-killed L. plantarum KU210152 increased brain-derived neurotrophic factor (BDNF) and tyrosine hydroxylase (TH) expression in HT-29 cells. In SH-SY5Y cells, pretreatment with L. plantarum KU210152 CM decreased Bax/Bcl-2 ratio and upregulated BDNF and TH expression. The CM inhibited caspase-9 and caspase-3 activities. The neuroprotective effects of L. plantarum KU210152 were also confirmed in fermented soy milk. Therefore, both L. plantarum KU210152 and the fermented soy milk can be used as functional ingredients with neuroprotective effects against oxidative stress.

Fluoride Determination in Milk, Soy, and Water-Based Products Using Ion-Selective Electrode and Direct Measurement Technique: Single-Laboratory Validation, First Action 2022.05.

BACKGROUND: The AOAC Stakeholder Panel on Infant Formula and Adult Nutritionals issued a call for methods for the determination of fluoride in infant formula. Standard Method Performance Requirement (SMPR®) 2014.016 was approved. OBJECTIVE: A single-laboratory validation (SLV) of a fluoride ion-selective electrode (F-ISE) method was completed. METHODS: Five powder samples were reconstituted, and hydrochloric acid was added to dissolve and release any bound fluoride. Samples included infant and adult nutritional products made from milk, whey, or soy, containing intact, partially hydrolyzed, or hydrolyzed proteins. Sodium citrate buffer was added to complex any interfering ions and adjust pH and ionic strength. Samples were analyzed in duplicate over 6 days using F-ISE. RESULTS: Results were calculated based on one of two least-squares (log fit) standard curves (0.02 to 0.1 µg/100 g or 0.1 to 2.0 µg/100 g). The LOQ for the method was determined to be 3.3 µg/100 g, which was far below the minimum analytical value of 30 µg/100 g required in the SMPR. The fluoride concentrations ranged from approximately 3 to 14 µg/100 g reconstituted powder and the intermediate precision, RSDr, ranged from 0.7 to 12.4%. Although all of these results are out of scope for the SMPR due to their low fluoride, it should be noted that the RSDr for formulas with fluoride concentrations ranging from approximately 9 to 14 µg/100 g ranged from 0.7 to 4.3%, and the RSDr for the two formulas with fluoride concentrations at or below the theoretical detection limit of the method were only approximately10 and 12.4%. Recoveries ranged from 94 to 98% for samples spiked at three levels with NIST-traceable standard solutions, meeting the SMPR. CONCLUSION: The F-ISE method meets the SMPR and was adopted as a First Action Official MethodSM. HIGHLIGHTS: The fluoride ion-selective electrode method presented here is affordable and easy to run. The addition of acid to the samples dissolves any minerals which may bind the fluoride.

Riboflavin bio-enrichment of soy beverage by selected roseoflavin-resistant and engineered lactic acid bacteria.

Some lactic acid bacteria (LAB) have the ability to synthesize riboflavin, a trait linked to the presence of ribG, ribB, ribA and ribH genes located in the rib operon. Previous screening of riboflavin producers identified several LAB strains belonging to different species with this ability, but none of them surpassed 0.25 mg/L production of the vitamin. In this study, we explored two strategies to obtain riboflavin-overproducing strains: by roseoflavin selection of mutants, and by the transformation of selected strains with plasmids pNZ:TuR.rib or pNZ:TuB.rib containing the genes ribG, ribB, ribA and ribH from Lactococcus cremoris MG1363. The resulting riboflavin-overproducing strains were able to produce yields between 0.5 and 6 mg/L in culture media and several of them were selected for the fermentation of soy beverages. Riboflavin in bio-enriched soy beverages was evaluated by direct fluorescence measurement and high-performance liquid chromatography-fluorescence analysis. Soy beverages fermented with the recombinant strains Lactococcus cremoris ESI 277 pNZ:TuB.rib and Lactococcus lactis INIA 12 pNZ:TuR.rib showed the highest riboflavin yields (>5 mg/L) after 24 h fermentation. On the other hand, roseoflavin-resistant mutant Limosilactobacillus fermentum INIA P143R2 was able to enrich fermented soy beverages with 1.5 mg/L riboflavin. Riboflavin-overproducing LAB strains constitute a good option for riboflavin enrichment of soy beverages by fermentation and the commercialization of such beverages could be very useful to prevent riboflavin deficiency.

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.

Quantification of the environmental impact arising from the utilization of whole and defatted Okara in fermentative and dehydration processes.

Okara is the insoluble pulp that remains after the grinding and filtration of soybeans during the production of soymilk and tofu. As it retains a noteworthy quantity of nutrients, there has been an increasing emphasis in the utilization of this residue for the development of sustainable processes. This study focused on assessing the environmental impact of employing okara as a medium for fermenting and dehydrating probiotic bacteria at laboratory scale. The evaluation was carried out using the Life Cycle Assessment (LCA) methodology, considering the entire process lifecycle. Whole okara and defatted okara were used as culture media for Lactiplantibacillus plantarum CIDCA 83114, followed by dehydration (either freeze-drying or spray-drying) and subsequent storage. For the purpose of comparison, both scenarios (whole and defatted okara) were evaluated using 1 kg of dehydrated final product for storage, as functional unit. Based on experimental results, the conservation of the received okara and the dehydration-storage (e.g., freezing and freeze-drying) phases were identified as the most significant environmental hotspots responsible for the most substantial impacts of the processes. The use of LCA facilitated the measurement of the environmental effects linked to the reutilization of okara as an agro-industrial residue, thus providing quantitative support when engineering its sustainable valorization.