The influence of α and α' subunits on SPI Pickering emulsions based on natural hybrid breeding varieties.

In this study, food-grade protein nanoparticles (Wild-NPs, α-lack-NPs, α'-lack-NPs, and (α + α')-lack-NPs) were organized as emulsion stabilizers via thermal induction. The effects of α and α' subunits in soybean protein isolate (SPI) on Wild nanoparticle Pickering emulsion (Wild-NPPEs), α-lack nanoparticle Pickering emulsion (α-lack-NPPEs), α'-lack nanoparticle Pickering emulsion (α'-lack-NPPEs) and (α + α')-lack nanoparticle Pickering emulsion ((α + α')-lack-NPPEs) were investigated. The Pickering emulsion stabilization mechanism indicated that the α'-lack-NPs particle size, surface hydrophobicity, and contact angle were mostly comparatively large. Therefore, the absence of the α' subunit made the desorption of protein nanoparticles at the oil and water interface require higher energy. Through the hydrophobic interaction between molecules, the structure and properties of the emulsion were improved, showing good stability. The existence of α'-lack-NPPEs leads to the formation of a gel-like network in the emulsion, which increases the viscosity of the emulsion and makes the network structure of the emulsion more uniform and denser.

The complexes of soybean protein isolate and procyanidin B2 have synergistic hypolipidemic activity at the cellular level by activating the AMPKα pathway.

The alkali method was used to prepare soybean protein isolate (SPI) and procyanidin B2 (PCB2) complexes, and the interaction between SPI and PCB2 was studied using multi-spectroscopic methods. The human hepatoma (HepG2) cell hyperlipidemia model was used to explore whether SPI-PCB2 has the potential for synergistic hypolipidemia. According to the findings, PCB2 was primarily linked to SPI via C-S and C-N bonds, and the addition of PCB2 reduced the α-helix structure content of SPI by 4.1%. At the cellular level, the optimal SPI-PCB2 ratio for lowering blood lipids was 1:1. Compared with the model group, the TG content and TC content in the 1:1 group were reduced by 28.7% and 26.3%, respectively. Western blot analysis revealed that SPI-PCB2 = 1:1 exerted synergistic hypolipidemic activity mainly by activating adenosine monophosphate-activated protein kinase α (AMPKα) phosphorylation, inhibiting 3-hydroxy-3-methylglutaryl CoA reductase (HMGCR) and fatty acid synthetase (FAS) protein expression, and upregulating carnitine palmitoyl transferase 1A (CPT1A) protein activity.

Optimization of culture medium and scale-up production of astaxanthin using corn steep liquor as substrate by response surface methodology.

Astaxanthin is a natural carotenoid with strong antioxidant activity. In this paper, the effects of carbon source, corn steep liquor, distiller grains, and initial pH on the growth and astaxanthin production of Phaffia rhodozyma D3 were evaluated. The optimal medium composition was 32 g/L glucose, 12 g/L corn steep liquor as nitrogen source, and the initial pH was 6.7. Phaffia rhodozyma D3 was cultured in a shake flask under these optimized conditions, the biomass was 6.47 g/L, the astaxanthin/OD475 was 15.16, and the astaxanthin content was 1.41 mg/g. The astaxanthin content was further increased to 4.70 mg/g by the combination of TiO2 stimulation and the expanding cultivation of P. rhodozyma D3 in a 5 L fermenter, which was 2.81 times that of the control group. Expanding fermentation implies the possibility of large-scale production in the astaxanthin industry. Corn steep liquor was used as an alternative nitrogen source to culture P. rhodozyma D3, which could both reduce the production cost of astaxanthin and increased the by-products utilization rate.

Optimization of fermentation for γ-aminobutyric acid (GABA) production by yeast Kluyveromyces marxianus C21 in okara (soybean residue).

γ-Aminobutyric acid (GABA) is a non-protein amino acid with a variety of physiological functions. Recently, yeast Kluyveromyces marxianus strains involved in the catabolism and anabolism of GABA can be used as a microbial platform for GABA production. Okara, rich in nutrients, can be used as a low-cost fermentation substrate for the production of functional materials. This study first proved the advantages of the okara medium to produce GABA by K. marxianus C21 when L-glutamate (L-Glu) or monosodium glutamate (MSG) is the substrate. The highest production of GABA was obtained with 4.31 g/L at optimization condition of culture temperature 35 °C, fermentation time 60 h, and initial pH 4.0. Furthermore, adding peptone significantly increased the GABA production while glucose and vitamin B6 had no positive impact on GABA production. This research provided a powerful new strategy of GABA production by K. marxianus C21 fermentation and is expected to be widely utilized in the functional foods industry to increase GABA content for consumers as a daily supplement as suggested.

Structural properties, antioxidant and immune activities of low molecular weight peptides from soybean dregs (Okara).

In this study, a method for preparing low molecular weight peptides (HPH-VAP) from okara using high-pressure homogenization assisted double enzymes was proposed. In order to explore its advantages, the effects of various methods on protein extraction rate and on the structure, antioxidant and immune properties of peptides were compared. The results showed that the protein extraction rate of this method was increased by 69% and 51% compared with other methods, and the structure only led to changes in the hydrogen bonds between peptide chains. HPH-VAP was screened out through functional characteristics, its structure was identified by HPLC-MS/MS, and further immunological activity analysis was carried out. The results showed that it promoted cell phagocytic ability, NO level and release of cytokines IL-6, IFN- γ, TNF-α. Therefore, this method is an effective and applicable method for industrial preparation of okara peptides, and has a positive effect on the reuse of okara resources.

Characteristics of lipoxygenase-based and lipoxygenase-deficient soy yogurt with modified okara.

Lipoxygenase-based and lipoxygenase-deficient okara were modified by Kluyveromyces marxianus fermentation, then adding modified okara back to the corresponding soymilk to prepare soy yogurt. The physicochemical properties, texture, and volatile components of soy yogurt were characterized. The results showed that okara modified by Kluyveromyces marxianus fermentation was rich in soluable dietary fiber and was imparted better water-holding capacity, swelling capacity, and oil-holding capacity. The soy yogurt with the modified okara was greatly enhanced in its appearance, texture and was relatively stable during storage. Moreover, lipoxygenase-based soy yogurt had a unique soybean flavor while lipoxygenase-deficient soy yogurt had a slight beany flavor and soybean flavor. This article guides a bio-modified method for okara and provides a theoretical basis for the further development and application of soy yogurt with high dietary fiber as well as lipoxygenase-deficient soy yogurt. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10068-021-01003-w.

Effect of pinolenic acid on oxidative stress injury in HepG2 cells induced by H2O2.

To investigate the effect and mechanism of pinolenic acid (PNA) on H2O2-induced oxidative stress injury in HepG2 cells. Methods: PNA was used to regulate oxidative stress injury of HepG2 cells induced by H2O2. Quantification of cell survival rate, accumulation of intracellular reactive oxygen species (ROS), and expression levels of anti-oxidation-related genes were determined using MTT, fluorescent probe technology (DCFH-DA), and real-time quantitative reverse transcription polymerase chain technology (qRT-PCR) method, respectively. Meanwhile, the activity of intracellular antioxidant enzymes was determined by biochemical methods. The results showed that PNA improved the survival rate of HepG2 cells induced by H2O2 (29.59%, high-dose group), reduced the accumulation of intracellular ROS (65.52%, high-dose group), and reduced the level of intracellular malondialdehyde (MDA; 65.52%, high-dose group). All these results were dose-dependent, which indicated that PNA can improve oxidative stress damage of cells. Furthermore, the mechanism of PNA regulating oxidative stress was investigated from the gene level. Results showed that under supplementation of PNA, the activity of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) had been improved (39.74%, 17.58%, and 23.83%, high-dose group). Further studies on gene expression which controls the activity of antioxidant enzymes showed that under the regulation of PNA, the expression level of Keap1 gene was decreased, while Nrf2 gene was increased. The expression levels of HO-1 and NQO1 in the downstream of Nrf2 were increased. Results indicated that under the regulation of PNA, Nrf2 was separated from Keap1, entered the nucleus, bound to ARE, and up-regulated the expression levels of HO-1 and NQO1 genes. Conclusion: PNA has a conspicuous improvement effect on oxidative stress damage induced by H2O2 in HepG2 cells. We also found the antioxidant mechanisms of PNA where it protected cells from oxidative stress damage by causing nuclear translocation of Nrf2 gene and up-regulated the expression levels of antioxidant enzymes in the downstream. This shows that PNA prevented oxidative stress by mediating the Keap1/Nrf2 transcriptional pathway and down-regulating enzyme activities.

Flavonoids derived from buckwheat hull can break advanced glycation end-products and improve diabetic nephropathy.

Diabetic nephropathy (DN) is the most important complication in patients with diabetes. The accumulation of advanced glycation end-products (AGEs) is the main reason for the development of DN. In this study, we investigated the mechanism of buckwheat hull flavonoids to break AGEs in vitro by measuring fluorescence analysis, three-dimensional fluorescence, protein molecular weight, free amino groups, and the sulfhydryl group content. Proteomics analysis was used to determine the effect of total buckwheat hull flavonoids (TBHF) intervention on protein differential expression in the kidney of db/db mice. The results showed that buckwheat hull flavonoids were potent in breaking AGEs in vitro, and they protected mice kidneys by regulating the renal AGE-RAGE pathway. This study lays a strong experimental and theoretical foundation for the development of new lysing agents to break AGEs. The findings should make an important contribution to the field of flavonoids in improving the application of diabetic nephropathy in the diet.

Production of ß-glucosidase from okara fermentation using Kluyveromyces marxianus.

The effective utilization of okara (soybean residue) has become a considerable challenge in recent years. In this paper, the potential advantages of ß-glucosidase production from okara fermented by Kluyveromyces marxianus were evaluated and the properties of the ß-glucosidase were also characterized. The results showed that okara can significantly induce the production of ß-glucosidase from K. marxianus. The ß-glucosidase activity was up to 4.5 U/mg under optimized fermentation conditions. The optimal parameters were as follows: fermentation temperature 35 °C, cultivation time 98 h, inoculum concentration 10%, and 30 g/L of okara. After two steps of purification using ammonium sulfate precipitation and Sephadex G-75 column chromatography, the activity of ß-glucosidase was 71.4 U/mg. The native enzyme was an approximately 66 kDa dimer consisting of two different subunits (22 and 44 kDa). The kinetic parameters of the K. marxianus ß-glucosidase, using pNPG as substrate, were V max 8.34 µmol min-1 mg-1 and K m 7.42 mM. The ß-glucosidase showed high thermostability and acid-alkali tolerance as well as low inhibition by DMSO (10-50%). In conclusion, this study supports the notion that okara fermentation by K. marxianus could be a useful process to produce ß-glucosidase.

Fermented ginseng improved alcohol liver injury in association with changes in the gut microbiota of mice.

The interactions among the liver, intestine and immune system play an important role in alcoholic liver injury. In this study, C57BL/6N mice with alcoholic injury were treated with unfermented and Lactobacillus fermentum KP-3-fermented ginseng. The indicators of hepatic steatosis, inflammation and injury were evaluated. The number of beneficial and harmful bacteria in the mice ileum and colon was counted by a traditional method; moreover, the diversity analysis of the cecum flora was performed. The alcohol exposure increased the levels of ALT, AST, TNF-α and IL-6 inflammatory factors and liver steatosis. In addition, the alcohol-fed miceexhibited a lower number of Lactobacilli and Bifidobacteria in the ileum and colon; the cecum flora diversity in the mice showed that alcohol obviously enhanced the abundance of the unclassified S24-7 of the Bacteroidetes phylum and the Proteobacteria genus of the Sutterella phylum and reduced the abundance of short-chain fatty acid-producing bacteria such as Akkermansia in the Verrucomicrobia phylum and those belonging to the Allobaculum genus, the Ruminococcus genus, and the Adlercreutzia genus in the Actinobacteria phylum. All these changes were improved by fermented ginseng. Conclusively, fermented ginseng could alleviate the alcoholic liver injury and disorder of the intestine by adjusting the intestinal flora.

Genome shuffling improved acid-tolerance and succinic acid production of Actinobacillus succinogenes.

Succinic acid is widely applied to chemical, pharmaceutical, food, and agricultural industries. With the rapid development of these industries, a great demand of succinic acid is required. The acid-tolerance and succinic acid production of Actinobacillus succinogenes strain were improved by using genome shuffling. Results showed that one modified strain AS-F32, with the best acid resistance and the highest succinic acid production, was obtained after 3 cycles of genome shuffling. The minimum growth pH of AS-F32 was 3.5, and the acid production and cell dry weight were 5.1 and 4.8 g/L in flask, improved 2.6 and 1.85 times over the start strain As-R2. Furthermore, the succinic acid yield of As-32 was 31.2 g/L and the dry cell weight was increased 44.4% by maintaining pH 4.8 with 7.0 M NH4OH in 5 L bioreactor, increased 1.1 times than the original strain As-R2.

Spectroscopic and molecular modeling approaches to investigate the interaction of bisphenol A, bisphenol F and their diglycidyl ethers with PPARα.

A fluorescence polarization (FP) assay for the simultaneous determination of bisphenol A (BPA), bisphenol F (BPF), bisphenol A diglycidyl ether (BADGE) and bisphenol F diglycidyl ether (BFDGE) was developed. The method was based on the competition between bisphenols (BPs) and fluorescein-labeled dexamethasone derivative (Dex-fl) for mouse peroxisome proliferator-activated receptor α ligand binding domain (mPPARα-LBD). A recombinant soluble protein derivative mPPARα-LBD* was prepared, then in vitro binding of 4 BPs to mPPARα-LBD* was investigated. Fluorescence polarization assay showed that these compounds exhibited different binding potencies with mPPARα-LBD*. Additionally, molecular dynamics simulations were performed to further understand the mechanism of BPs binding affinity for mPPARα-LBD*. Docking results elucidated that the driving forces for the binding of BPs to mPPARα-LBD* were predominantly dependent on hydrophobic and hydrogen-bonding interactions. Comparison of the calculated binding energies vs. experimental binding affinities yielded a good correlation (R2 = 0.7258). The proposed method has potential for multi-residue detection of BPA, BPF, BADGE, and BFDGE.