A new biostimulant derived from soybean by-products enhances plant tolerance to abiotic stress triggered by ozone.

BACKGROUND: Tropospheric ozone is an air pollutant that causes negative effects on vegetation, leading to significant losses in crop productivity. It is generated by chemical reactions in the presence of sunlight between primary pollutants resulting from human activity, such as nitrogen oxides and volatile organic compounds. Due to the constantly increasing emission of ozone precursors, together with the influence of a warming climate on ozone levels, crop losses may be aggravated in the future. Therefore, the search for solutions to mitigate these losses becomes a priority. Ozone-induced abiotic stress is mainly due to reactive oxygen species generated by the spontaneous decomposition of ozone once it reaches the apoplast. In this regard, compounds with antioxidant activity offer a viable option to alleviate ozone-induced damage. Using enzymatic technology, we have developed a process that enables the production of an extract with biostimulant properties from okara, an industrial soybean byproduct. The biostimulant, named as OEE (Okara Enzymatic Extract), is water-soluble and is enriched in bioactive compounds present in okara, such as isoflavones. Additionally, it contains a significant fraction of protein hydrolysates contributing to its functional effect. Given its antioxidant capacity, we aimed to investigate whether OEE could alleviate ozone-induced damage in plants. For that, pepper plants (Capsicum annuum) exposed to ozone were treated with a foliar application of OEE. RESULTS: OEE mitigated ozone-induced damage, as evidenced by the net photosynthetic rate, electron transport rate, effective quantum yield of PSII, and delayed fluorescence. This protection was confirmed by the level of expression of genes associated with photosystem II. The beneficial effect was primarily due to its antioxidant activity, as evidenced by the lipid peroxidation rate measured through malondialdehyde content. Additionally, OEE triggered a mild oxidative response, indicated by increased activities of antioxidant enzymes in leaves (catalase, superoxide dismutase, and guaiacol peroxidase) and the oxidative stress index, providing further protection against ozone-induced stress. CONCLUSIONS: The present results support that OEE protects plants from ozone exposure. Taking into consideration that the promotion of plant resistance against abiotic damage is an important goal of biostimulants, we assume that its use as a new biostimulant could be considered.

Effects of fermentation with different probiotics on the quality, isoflavone content, and flavor of okara beverages.

The present study aimed to prepare and evaluate a new probiotic functional beverage, using single-probiotic and compound probiotic fermentation on okara. Four different forms of fermentation microorganisms used were Lacticaseibacillus rhamnosus S24 (Lr), Lacticaseibacillus paracasei 6244 (Lp), Lactobacillus acidophilus 11,073 (La), and mixed fermentation (Lr + Lp + La). The physicochemical properties, antioxidant activity, flavor change, and storage period of fermented okara beverages with probiotics were investigated. The results showed that different fermentation schemes could significantly improve the physicochemical properties, antioxidant activity, and sensory quality of the okara beverages. The number of viable bacteria in the Lp group (3.53 × 108 CFU/mL), isoflavone content (0.514 µg/mL) were the highest; total phenol and flavonoid content were 3.32 and 5.68 times higher than in the CK group, respectively. DPPH and ABTS+ free radical scavenging rates were increased by 11.32% and 20%, respectively (p < .05). Through SPME/GC-MS analysis, 44 volatile compounds were identified in the Lr + Lp + La groups, mainly as a result of changes in alcohols and aldehydes produced by fermentation metabolism. It enhances the floral and fruity aroma of the okara beverage. All probiotic-fermented okara beverages can be stored at 4°C for 15 days, with probiotic activity greater than 107 CFU/mL. This study can obtain a probiotic okara beverage rich in soybean isoflavones and with good flavor. Overall, okara can be used to develop functional beverages containing probiotics and contribute to a zero-waste approach in the food industry.

Relationship Between the Frequency of Bowel Movements and Fecal Bacteroides in Japanese Women.

The intestinal microbiota is involved in many diseases, such as constipation, obesity, and inflammatory bowel disease. To determine the associations between the gut microbiome and the frequency of bowel movements, we performed cross-sectional correlation analysis at the baseline and longitudinal correlation analysis after the intervention. Forty-three women were enrolled in this study. All participants ingested soymilk-okara powder (15 g) daily for 12 weeks. They recorded the ingested okara powder amounts and their frequency of bowel movements during the entire 12 weeks of the intervention. The fecal microbiota percentages were measured at baseline and after 12 weeks of intervention. Two women who did not completely record the frequency of bowel movements were excluded. In the cross-sectional correlation analysis at the baseline, there was a significant positive correlation between the relative abundance of the Bacteroides genus in the feces and the frequency of bowel movements per week (R = 0.429, p = 0.005) and a significant negative correlation between the relative abundance of Clostridium cluster XI in the feces and the frequency of bowel movements per week (R = -0.315, p = 0.045). Moreover, in the longitudinal correlation analysis, the difference in the relative abundance of Bacteroides genus in feces between the baseline and after the intervention significantly correlated with the changes in the frequency of bowel movements per week (R = 0.492, p = 0.001). Therefore, it was suggested that there was a relationship between the gut relative abundance of the Bacteroides genus and the frequency of bowel movements.

Extraction of insoluble soybean fiber by alternating ultrasonic/alkali and its improved superior physicochemical and functional properties.

Okara, as a by-product of soybean processing, is rich in insoluble dietary fiber (IDF), which is a carbohydrate polymer with various insoluble polysaccharides. Nowadays, the extraction of IDF with excellent functional properties has become a research hotspot. In this work, we further proposed an alternating alkali/ultrasound method for the efficient extraction of IDF. The sequential treatments of alkali (A-ISF), alkali-ultrasonic (AU-ISF), ultrasonic-alkali (UA-ISF), ultrasonic-alkali-ultrasonic (UAU-ISF) and alkali-ultrasonic-alkali (AUA-ISF) were applied to extract insoluble soybean fiber (ISF). FTIR and XRD results proved the typical structure of ISFs, and TGA results demonstrated the improved thermal stability of UAU-ISF and AUA-ISF. Chemical composition measurement showed that UAU-ISF and AUA-ISF exhibited higher cellulose content (>83 %). SEM results revealed that ultrasonic treatment led to a decomposition of okara matrix and significant porous structure in ISFs with an amplified collapse effect, resulting in an increase of the pore size of ISFs, and strengthening the properties of UAU-ISF and AUA-ISF in higher water (>15 g/g)/oil (>12 g/g) holding capacities, cholesterol binding capacity (>36 mg/g), and cation exchange capacity (>0.3 mmol/g), thus providing new insights for the preparation of ISF with high functional properties that are beneficial for human intestinal health.

Metabolomics analysis of okara probiotic beverages fermented with Lactobacillus gasseri and Limosilactobacillus fermentum by LC-QTOF-MS/MS.

In this study, okara was fermented with probiotic strains Lactobacillus gasseri LAC 343 and Limosilactobacillus fermentum PCC, respectively. Significant increases in cell count (by 2.22 log CFU/mL for LAC and 0.82 log CFU/mL for PCC) and significant decreases in pH (by 1.31 for LAC and 1.03 for PCC) were found in fermented okara slurry. In addition, strain LAC tended to produce amino acids, while strain PCC depleted most amino acids. An untargeted metabolomic-based approach using liquid chromatography coupled with quadrupole time-of-flight mass spectrometry was used to further understand the compositional changes and potential health benefits by identifying bioactive metabolites in fermented okara slurry. We successfully identified various beneficial bioactive compounds including γ-aminobutyric acid, indolelactic acid, d-phenyllactic acid, and p-hydroxyphenyllactic acid which had differences in fold-changes in okara slurry fermented with different strains. Our study indicated the feasibility of using probiotics to ferment okara for novel functional food development.

From macro- to nano- scales: Effect of fibrillary celluloses from okara on performance of edible starch film.

Starch/cellulose composite is one of the most promising systems since both matrix and reinforce agent have same chemical unite glucose, which results in an excellent compatibility. In this work, edible starch film was developed by compositing starch with diverse fibrillary celluloses (FCs) derived from okara, employing a confluence of chemical interactions and mechanical influences. Since diameter of the FCs can be easily controlled by processing methodologies, it is the first time to systematically investigate the effect of diameter of the FCs from macro to nano-scales on the performances of starch-based film. The fabricated macro- and nano-fibrillar celluloses and reinforced starch films were characterized by scanning electron microscope, optical microscopy, Fourier transform infrared spectroscopy, Rheometer and contact angle. Results showed that the FCs increased modulus (about 170 %) and tensile strength (about 180 %) significantly as expected since they are well-compatible and some chemical interactions. It was found that nano-fibrillary celluloses (CNFs) improve the toughness (about 20 %) of the starch film more efficiently, which improved the well-recognized weakness of starch-based materials. The nano-scale roughness on the surface of the starch film caused by different shrinkage ratios between starch and CNFs during drying reduced water sensitivity, which is another well-recognized weakness of starch film.

Effect of modified okara insoluble dietary fibre on the quality of yoghurt.

The aim of this study was to investigate the potential of adding soya bean dregs insoluble dietary fibre (IDF) modified by jet cavitation combined with cellulase to yoghurt to improve its functional properties (Yoghurt was prepared by adding 10 µL of yoghurt fermenter to 100 mL of milk, fermented to pH 4.5 in a constant temperature incubator at 42 °C, and then stored in a refrigerator at 4 °C after adding IDF separately). The results showed that the modified IDF had a rough structure with high water-holding capacity and sodium cholate adsorption capacity. The addition of modified IDF improved the pH, hardness, and elasticity of the yoghurt. During the entire storage period, the titratable acidity and whey precipitation rate of the modified IDF yoghurt gradually increased, and antioxidant activity gradually decreased, and its titratable acidity, whey precipitation rate, and antioxidant activity had a significant advantage compared with those of the blank group yoghurt. In conclusion, the modified soya bean dregs IDF-added yoghurt prepared by jet cavitation combined with the cellulase method has the potential for sodium cholate adsorption capacity and antioxidant activity, which can confer unique functional properties and improve the pH, texture, and reduce whey precipitation of yoghurt. This study provides a scientific basis for the application of soya bean dregs IDF as a fibre fortifier in yoghurt production and suggests innovative ideas for the design of functional dairy products.

Ultrasound-assisted fermentation for antioxidant peptides preparation from okara: Optimization, stability, and functional analyses.

This study investigated the effects of ultrasound-assisted fermentation (UAF) on the preparation of antioxidant peptides (UAFP) from okara and examined their content, chemical structures, and antioxidant activity. After the optimal ultrasonic processing (time, 20 min; frequency, 45 KHz; power, 120 W/L), the peptide content yield reached the maximum of 12.36 ± 0.02 mg/mL, and their DPPH free radical scavenging rate was 65.15 ± 0.32 %. UAF increased the number of globular aggregates with deeper gullies, a looser structure, and higher porosity. The experiments conducted using the oxidative stress injury model of HepG2 cells showed that okara UAFP promoted cell growth and exerted a protective effect. Moreover, ultrasonic treatment remarkably improved the environmental stability (NaCl, glucose, sodium benzoate, temperature, pH, metal ions) and antioxidant activity of UAFP. Concisely, optimal ultrasonic processing can aid the fermentation of agroindustrial by-products to prepare antioxidant peptides, such as natural food antioxidant peptides from soybean waste.

Fabrication of composite hydrogels by sonication-assisted assembly of okara cellulose nanofibers and chitosan: structure and properties.

BACKGROUND: Okara cellulose is a highly abundant, green, sustainable, and biodegradable polymer with many potential industrial applications. In this study, we fabricated composite hydrogels with okara cellulose nanofibers (CNFs) and chitosan (CH) by hydrating, sonicating, and heating them at 100 °C for 30 min, and then induced their assembly by cooling. The effects of okara CNF (with and without 2,2,6,6-tetramethylpiperidinyloxy (TEMPO) oxidation) and CH concentration on the structure and properties of the hydrogels was examined, including their microstructure, surface properties, rheological properties, and thermal stability. RESULTS: Our results indicate that there was an electrostatic attraction between the anionic okara CNF and cationic CH, which facilitated hydrogel formation. The surface, textural, rheological, and thermal stability properties were better for the composite hydrogels than for the single CH ones, as well as for the CNF that had undergone TEMPO oxidation. For the TC-CH hydrogels, the contact angle was 39.5°, the interfacial tension was 69.1 mN m-1 , and the surface tension was 1.44 mN m-1 . CONCLUSION: In this study, the novel hydrogels developed may be useful as a soft material in a range of applications in foods, supplements, health care products, cosmetics, and drugs. © 2023 Society of Chemical Industry.

Composite hydrogels formed from okara cellulose nanofibers and carrageenan: Fabrication and characterization.

Currently, there is great interest in converting edible agro-waste, such as okara from soybean production, into value-added products. For this study, we focus on fabricating composite hydrogels from okara cellulose nanofibers (CNFs) and carrageenan (CA). We also examined the effects of TEMPO oxidation of the okara CNFs, as well as CA concentration, on the microstructure and physicochemical properties of the composite hydrogels. The water holding capacity, oil holding capacity, surface tension, gel strength, and viscoelasticity of the composite microgels increased with increasing CA concentration, and it was found that the highest values were obtained for TC-CA2 hydrogel: contact angle = 43.6° and surface tension = 45.12 mN/m, which was attributed to the formation of a more regular and dense three-dimensional gel network. All the CNF-CA microgels had highly anionic ζ-potential values (-38.8 to -50.1 mV), with the magnitude of the negative charge increasing with TEMPO oxidation and carrageenan concentration. These results suggest there would be strong electrostatic repulsion between the composite hydrogels. The composite microgels produced in our work may be useful functional materials for utilization within the food industry, thereby converting a waste product into a valuable commodity.

High-Pressure Pasteurization of Oat Okara.

The issue of the short microbiological shelf life of residues from the plant-based beverage industry creates a large food waste problem. Today, the oat beverage residue, in this study referred to as oat okara, is generally converted to energy or used as animal feed. High-pressure pasteurization (200 MPa, 400 MPa, and 600 MPa) was applied to oat okara to investigate the effect on shelf life and microbiological activity. A 4-week microbiological storage study was performed and thermal properties, viscosity, and water and oil holding capacities were analyzed. The total aerobic count, including yeast and mold, was significantly reduced (p < 0.05) by 600 MPa after four weeks of storage at 4 °C. The content of lactic acid bacteria after four weeks of storage was low for untreated oat okara (3.2 log CFU/g) but, for 600 MPa, the content remained at the detection limit (2.3 log CFU/g). Conversely, the treatments of 200 MPa and 400 MPa increased the microbial content of the total aerobic count significantly (p < 0.05) after two weeks in comparison to untreated oat okara. The thermal properties of untreated and high-pressure-treated oat okara demonstrated an increase in protein denaturation of the 12S globulin, avenalin, when higher pressure was applied (400-600 MPa). This was also confirmed in the viscosity measurements where a viscosity peak for avenalin was only present for untreated and 200 MPa treated oat okara. The water holding capacity did not change as a function of high-pressure treatment (3.5-3.8 mL/g) except for the treatment at 200 MPa, which was reduced (2.7 mL/g). The oil holding capacity was constant (1.2-1.3 mL/g) after all treatments. High-pressure pasteurization of 600 MPa reduced the microbial content in oat okara resulting in a shelf life of 2-4 weeks. However, more research is required to identify the microorganisms in oat okara to achieve a microbiologically safe product that can be used for food applications.

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.

High-Pressure Pasteurization of Soy Okara.

Okara is a by-product from the production of soy beverages, which has a high content of protein and fiber. Even though it has a high nutritional value, it is generally discarded or used as animal feed or compost. The problem is its short shelf life due to its high water content and high water activity. The aim of this study was to investigate the effect of high-pressure pasteurization at 200 MPa, 400 MPa, and 600 MPa on the shelf life of soy okara. Microbiological growth, as well as thermal properties, viscosity, water holding capacity, and oil holding capacity, was evaluated after the pressure treatments. Treatment at 600 MPa significantly reduced (p < 0.05) the growth of total aerobic count, yeast and mold, and lactic acid bacteria for up to four weeks of storage at 4 °C. The pasting properties were increased while the water and oil holding capacities of the soy okara did not significantly change (p > 0.05) after high-pressure pasteurization at 400 MPa and 600 MPa. High-pressure pasteurization is therefore a potential application technique for soy okara to produce a microbiologically safe product with maintained functional properties. However, more research is needed to optimize the process and to further investigate the microbiological species present in untreated soy okara to exclude any potential food safety risks.

Impacts of Ultrasonic Treatment for Black Soybean Okara Culture Medium Containing Choline Chloride on the ß-Glucosidase Activity of Lactiplantibacillus plantarum BCRC 10357.

The effects of ultrasonic treatment for the culture medium of solid black soybean okara with choline chloride (ChCl) on the survival and ß-glucosidase activity of Lactiplantibacillus plantarum BCRC 10357 (Lp-BCRC10357) were investigated. A mixture of 3% dried black soybean okara in de Man-Rogosa-Sharpe (w/v) was used as the Oka medium. With ultrasonic treatment (40 kHz/300 W) of the Oka medium at 60 °C for 3 h before inoculation, the ß-glucosidase activity of Lp-BCRC10357 at 12 h and 24 h of incubation amounted to 13.35 and 15.50 U/mL, respectively, which was significantly larger than that (12.58 U/mL at 12 h and 2.86 U/mL at 24 h) without ultrasonic treatment of the medium. This indicated that ultrasonic treatment could cause the microstructure of the solid black soybean okara to be broken, facilitating the transport of ingredients and Lp-BCRC10357 into the internal structure of the okara for utilization. For the effect of ChCl (1, 3, or 5%) added to the Oka medium (w/v) with ultrasonic treatment before inoculation, using 1% ChCl in the Oka medium could stimulate the best response of Lp-BCRC10357 with the highest ß-glucosidase activity of 19.47 U/mL in 12 h of incubation, showing that Lp-BCRC10357 had a positive response when confronting the extra ChCl that acted as an osmoprotectant and nano-crowder in the extracellular environment. Furthermore, the Oka medium containing 1% ChCl with ultrasonic treatment led to higher ß-glucosidase activity of Lp-BCRC10357 than that without ultrasonic treatment, demonstrating that the ultrasonic treatment could enhance the contact of ChCl and Lp-BCRC10357 to regulate the physiological behavior for the release of enzymes. In addition, the analysis of the isoflavone content and antioxidant activity of the fermented product revealed that the addition of 1% ChCl in the Oka medium with ultrasonic treatment before inoculation allowed a higher enhancement ratio for the biotransformation of isoflavone glycosides to their aglycones, with a slight enhancement in the antioxidant activity at 24 h of fermentation. This study developed a methodology by combining ultrasonic treatment with a limited amount of ChCl to allow the culture medium to acclimate Lp-BCRC10357 and release high levels of ß-glucosidase, and this approach has the potential to be used in the fermentation of okara-related products as nutritional supplements in foods.

Production, structural and functional characteristics of soluble dietary fiber from fermented okara by Penicillium expansum.

Soluble dietary fiber (SDF), an important prebiotic, has attracted growing attention, due to its great health effects and wide application. This study focused on the preparation of SDF from fermented okara. The yield of SDF obtained through Penicillium expansum fermentation (FSDF) reached 45.63 % (w/w) under the optimal conditions (pH 6.7, inoculum size 9.5 %, and time 29 h) by response surface methodology, which were 1.92 and 4.43 times higher than those of phosphate-citric acid treatment and untreated okara. Infrared spectra and X-ray diffraction indicated that three SDFs had similar spectral distribution and crystalline region. Moreover, FSDF displayed looser and more porous microstructures. Meanwhile, the composition ratio of monosaccharides has changed. FSDF exhibited higher water solubility (97.46 %), glucose adsorption capacity (203.73 mg/g), sodium cholate adsorption capacity (13.07 mg/g), cholesterol adsorption capacity (6.69- 7.62 mg/g) and radical (ABTS+, hydroxyl and DPPH) scavenging capacity. Additionally, three SDFs didn't degrade by upper gastrointestinal tract and could improve the proportion of beneficial intestinal flora in vitro, such as Lactobacillus and Bifidobacterium. Overall, the FSDF prepared in this study was a functional ingredient with great potential in foods.

Modeling the Influence of Okara Flour Supplementation from Time-Temperature Drying Treatment on the Quality of Gluten-Free Roll Produced from Rice Flour.

Okara, an unassuming residue, is emerging as a notable reservoir of essential nutrients, encompassing an abundant supply of protein, dietary fiber, and potent antioxidant components. Hence, the incorporation of okara as an ingredient in the production of rice flour-based rolls held a considerable interest in nutritional and functional aspects. Okara flour supplement was prepared by drying at 100 °C for 2 h and selected based on the highest antioxidant level. Gluten-free rolls were prepared containing 0, 5, and 10% okara flour dried at 100 °C for 2 h, and their physicochemical properties were analyzed. Okara flour addition reduced the deformation of gluten-free batter roll by improving solid and liquid-like behavior, as evaluated with rheological measurements. Rolls containing okara flour processed at 100 °C for 2 h had increased firmness and decreased specific volume compared to the control. However, there were no significant differences in the sensory evaluation scores, suggesting that the consumers' acceptance of the control and the Okara rolls was similar. Okara flour supplement at 10% addition led to the nutritional improvement of the gluten-free rolls (increase of 2.4% protein and 1.32 times dietary fiber, dry basis). In contrast, there were no significant differences in the antioxidant level compared to the control. Okara flour is a functional ingredient with potential use in gluten-free products and a variety of novel products where enrichment is desirable.

Insoluble Dietary Fiber from Okara Combined with Intermittent Fasting Treatment Synergistically Confers Antiobesity Effects by Regulating Gut Microbiota and Its Metabolites.

Insoluble dietary fiber (IDF) was recently revealed to have an antiobesity impact. However, the impact and potential mechanism of high-purity IDF derived from okara (HPSIDF) on obesity caused by a high-fat diet (HFD) remain unclear. Except for dietary supplementation, intermittent fasting (IF) has attracted extensive interest as a new dietary strategy against obesity. Thus, we hypothesize that HPSIDF combined with IF treatment may be more effective in preventing obesity. In this study, HPSIDF combined with IF treatment synergistically alleviated HFD-induced dyslipidemia, impaired glucose homeostasis, systemic inflammation, and fat accumulation. Furthermore, gut microbiota dysbiosis and lowered short-chain fatty acid synthesis were recovered by HPSIDF combined with IF treatment. Meanwhile, metabolomic analysis of feces revealed that HPSIDF combined with IF treatment obviously reversed the alterations of metabolic pathways and differential metabolites induced by HFD, which were linked to the modulations of the gut microbiota. Collectively, our findings indicated that HPSIDF combined with IF treatment has great potential to substantially enhance antiobesity efficacy by modulating the gut microbiota and its metabolites.

The Effects of Okara Ratio and Particle Size on the Physical Properties and Consumer Acceptance of Tofu.

Okara, the solid byproduct of soymilk production, poses a sustainability concern, despite being rich in fiber and other healthful compounds. In this study, the physical properties of tofu made from soymilk fortified with differing levels of okara-either whole or fine (<180 µm)-and made with the traditional coagulant nigari were examined. The yield increased linearly with the okara concentration with values of 18.2-29.5% compared to 14.5% for the control. The initial moisture in the fortified samples was higher than the control (79.69-82.78% versus 76.78%), and both the expressible moisture and total moisture after compression were also greater in the fortified samples. With a few exceptions, the texture parameters did not differ between samples. Dynamic rheology showed that all samples had G' > G″. The storage moduli increased at different rates during each gelling step, with G' before and after gelling increasing with the fortification level, and was greater for the samples with fine particles than with whole particles. Consumer sensory panels using the hedonic scale showed traditional tofu had a slightly higher acceptability, but the panelists indicated they would be more willing to purchase okara-fortified tofu because of the health and sustainability benefits it might have. Thus, tofu could be produced with added okara with predictable but not profound changes in its physical properties.

Evaluation of Antioxidant Activities from a Sustainable Source of Okara Protein Hydrolysate Using Enzymatic Reaction.

Okara is a solid byproduct created during the processing of soy milk. The production of protein hydrolysates utilizing enzymatic tests such as papain can result in the production of bioactive peptides (BPs), which are amino acid sequences that can also be produced from the okara protein by hydrolysis. The objective of this study was to investigate the antioxidant activities of okara hydrolysates using papain, based on the in silico and in vitro assays using the papain enzyme. We found that using the in silico assessment, the antioxidant peptides can be found from the precursor (glycinin and conglycinin) in okara. When used as a protease, papain provides the maximum degree of hydrolysis for antioxidative peptides. The highest-peptide-rank peptide sequence was predicted using peptide ranks such as proline-histidine-phenylalanine (PHF), alanine-aspartic acid-phenylalanine (ADF), tyrosine-tyrosine-leucine (YYL), proline-histidine-histidine (PHH), isoleucine-arginine (IR), and serine-valine-leucine (SVL). Molecular docking studies revealed that all peptides generated from the parent protein impeded substrate access to the active site of xanthine oxidase (XO). They have antioxidative properties and are employed in the in silico approach to the XO enzyme. We also use papain to evaluate the antioxidant activity by using in vitro tests for protein hydrolysate following proteolysis. The antioxidant properties of okara protein hydrolysates have been shown in vitro, utilizing DPPH and FRAP experiments. This study suggests that okara hydrolysates generated by papain can be employed as natural antioxidants in food and for further applications, such as active ingredients for antioxidants in packaging.

Label-free quantitative proteomics reveals the potential mechanisms of insoluble dietary fiber from okara in improving hepatic lipid metabolism of high-fat diet-induced mice.

The high purity insoluble dietary fiber (IDF) from okara is a natural component with a potentially positive effect on a high-fat diet (HFD)-induced hepatic metabolic disorders, although its regulatory mechanism remains unclear. This study aims to elucidate the potential pathways and key proteins of IDF for the amelioration of hepatic lipid metabolism in mice fed with HFD. Here, we used label-free quantitative proteomics technology to quantity and identify differentially expressed proteins in the liver that are associated with IDF treatment. The differentially expressed proteins were assessed by GO annotation and KEGG pathways. Western blot and qRT-PCR analyses were conducted to validate the potential targets regulated by IDF. In total, 73 differentially expressed proteins were identified, of which 27 were up-regulated (FC > 1.5) and 46 were down-regulated (FC < 0.667). GO analysis suggested that differentially expressed proteins were mainly located in the cell and organelles, regulated biological processes, and were associated with enzyme activity and molecular binding. The KEGG pathway enrichment analysis further demonstrated glycolysis/gluconeogenesis, pyruvate metabolism, TCA cycle, arginine biosynthesis, alanine, aspartate and glutamate metabolism, and retinol metabolism were affected. The combination of proteomics, Western blot, and qRT-PCR suggested that ACS, ACLY, GOT1, GLS2, NAGS, CYP4A10, CYP3A25, and CYP2A5 in these pathways might be key proteins for IDF intervention. Taken together, our findings elucidate new mechanisms involved in how IDF affects hepatic metabolism, provide important information for the functional food industries, and improve the added value of okara. SIGNIFICANCE: Okara is evidenced as a high-quality by-product with several nutritional components, especially dietary fiber (50-60%) labeled as "The Seventh Nutrient". Previous studies have shown that IDF has a positive potential effect on a high-fat diet (HFD)-induced hepatic metabolic disorders, but its molecular mechanism remains unclear. To elucidate the therapeutic mechanism of IDF at the protein level, a label-free quantitative proteomic analysis was used to identify the dynamic changes of the liver proteome between HIDF and HFD groups in this study. These results provide a new perspective for exploring the therapeutic mechanism of IDF at the protein level and enlightenment for promoting the comprehensive utilization of okara.