Bioconversion of lignans in flaxseed cake by fermented tofu microbiota and isolation of Enterococcus faecium strain ZB26 responsible for converting secoisolariciresinol diglucoside to enterodiol.

Human intestinal microbiota plays a crucial role in converting secoisolariciresinol diglucoside, a lignan found in flaxseed, to enterodiol, which has a range of health benefits: antioxidative, antitumor, and estrogenic/anti-estrogenic effects. Given the high secoisolariciresinol diglucoside content in flaxseed cake, this study investigated the potential of co-fermenting flaxseed cake with fermented soybean product to isolate bacterial strains that effectively convert secoisolariciresinol diglucoside to enterodiol in a controlled environment (in vitro). The co-fermentation process with stinky tofu microbiota significantly altered the lignan, generating 12 intermediate lignan metabolites as identified by targeted metabolomics. One particular promising strain, ZB26, demonstrated an impressive ability to convert secoisolariciresinol diglucoside. It achieved a conversion rate of 87.42 ± 0.33%, with secoisolariciresinol and enterodiol generation rates of 94.22 ± 0.51% and 2.91 ± 0.03%, respectively. Further optimization revealed, under specific conditions (0.5 mM secoisolariciresinol diglucoside, pH 8, 30 °C for 3 days), ZB26 could convert an even higher percentage (97.75 ± 0.05%) of the secoisolariciresinol diglucoside to generate secoisolariciresinol (103.02 ± 0.16%) and enterodiol (3.18 ± 0.31%). These findings suggest that the identified strains ZB26 have promising potential for developing functional foods and ingredients enriched with lignans.

Anti-Inflammatory Effects of Barley Sprout Fermented by Lactic Acid Bacteria in RAW264.7 Macrophages and Caco-2 Cells.

The anti-inflammatory effects of supernatants produced from sprouted barley inoculated with Lactiplantibacillus plantarum KCTC3104 (Lp), Leuconostoc mesenteroides KCTC3530 (Lm), Latilactobacillus curvatus KCTC3767 (Lc), or a mixture of these lactic acid bacteria were investigated using RAW264.7 macrophages. BLp and BLc, the lyophilized supernatants of fermented sprouted barley inoculated with Lp and Lc, respectively, effectively reduced the nitric oxide (NO) levels hypersecreted by lipopolysaccharide (LPS)-stimulated RAW264.7 and LPS-stimulated Caco-2 cells. BLp and BLc effectively reduced the NO levels in LPS-stimulated RAW264.7 macrophages, and these effects tended to be concentration-dependent. BLc and BLp also exhibited strong DPPH radical scavenging activity and immunostimulatory effects. BLp and BLc significantly suppressed the levels of NO and pro-inflammatory cytokines such as TNF-α, IL-1ß, and IL-6 in LPS-stimulated RAW264.7 macrophages and LPS-stimulated Caco-2 cells, indicating their anti-inflammatory effects. These effects were greater than those of unfermented barley sprout (Bs). The functional components of Bs, BLp, and BLc were analyzed by HPLC, and it was found that lutonarin and saponarin were significantly increased in the fermented sprouted barley sample inoculated with Lp and Lc (BLp and BLc).

Mycoproteins and their health-promoting properties: Fusarium species and beyond.

Filamentous fungal mycoproteins have gained increasing attention as sustainable alternatives to animal and plant-based proteins. This comprehensive review summarizes the nutritional characteristics, toxicological aspects, and health-promoting effects of mycoproteins, focusing on those derived from filamentous fungi, notably Fusarium venenatum. Mycoproteins are characterized by their high protein content, and they have a superior essential amino acid profile compared to soybeans indicating excellent protein quality and benefits for human nutrition. Additionally, mycoproteins offer enhanced digestibility, further highlighting their suitability as a protein source. Furthermore, mycoproteins are rich in dietary fibers, which have been associated with health benefits, including protection against metabolic diseases. Moreover, their fatty acids profile, with significant proportions of polyunsaturated fatty acids and absence of cholesterol, distinguishes them from animal-derived proteins. In conclusion, the future of mycoproteins as a health-promoting protein alternative and the development of functional foods relies on several key aspects. These include improving the acceptance of mycoproteins, conducting further research into their mechanisms of action, addressing consumer preferences and perceptions, and ensuring safety and regulatory compliance. To fully unlock the potential of mycoproteins and meet the evolving needs of a health-conscious society, continuous interdisciplinary research, collaboration among stakeholders, and proactive engagement with consumers will be vital.

A two-step method of cyclolinopeptide (linusorb) preparation from flaxseed cake via dry-screening.

Linusorbs (LO), cyclolinopeptides, are a group of cyclic hydrophobic peptides and considered a valuable by-product of flaxseed oil due to numerous health benefits. Currently applied acetone or methanol extraction could contaminate the feedstocks for further food-grade application. Using flaxseed cake as feedstock, this study established a practical method for preparing LO from pressed cake. Firstly, LO composition of 15 flaxseed cultivars was analyzed. Next, cold-pressed cake was milled and screened mechanically. The kernel and hull fractions were separated based on the disparity of their mechanical strength. Monitored by hyperspectral fluorescence, the LO-enriched kernel fraction separated from cold-pressed flaxseed cake was further used as feedstock for LO production. After ethanol extraction, partition, and precipitation, LOs were extracted from cold-pressed flaxseed cake with a purity of 91.4%. The proposed method could serve as feasible flaxseed cake valorization strategy and enable the preparation of other polar compounds such as flax lignan and mucilage.

Stability of tryptophan-containing LOs in flaxseed oil and their response towards γ-tocopherol.

Linusorbs (LOs), significantly influence oil quality and sensory properties of flaxseed oil. Trp-containing LOs exhibit distinct oxidative behavior when γ-tocopherol (γ-T) is present. Polar fractions of crude flaxseed oil were stripped via silica absorption, and reintroduced (LO and γ-T) separately into the oil matrix to investigate their interaction during storage. Compared with crude oil, LOs account for 18.49% reduction of p-anisidine value, while LOs with γ-T contributed to most of the endogenous antioxidant effect in crude oil. γ-T was found to suppress oxidation of Trp-containing LO at early stage (Met form), while facilitate oxidation while at their mid-stage (MetO form, Methionine sulfoxide). In vitro oxidation shows that CLD more likely cleaved into peptide fragments, while few products retain intact ring structures. LC-MS/MS analysis and silicon simulation revealed proximity between MetO and Trp residues, facilitating inter- or intra-molecular reactions and ring structure rupture. Remarkably, the presence of γ-T facilitate these phenomena.

A comparative study on flaxseed lignan biotransformation through resting cell catalysis and microbial fermentation by ß-glucosidase production Lactiplantibacillus plantarum.

BACKGROUND: Flax lignan has attracted much attention because of its potential bioactivities. However, the bioavailability of secoisolariciresinol diglucoside (SDG), the main lignan in flaxseed, depends on the bioconversion by the colon bacteria. Lactic acid bacteria (LAB) with ß-glucosidase activity has found wide application in preparing bioactive aglycone. RESULTS: LAB strains with good ß-glucosidase activity were isolated from fermented tofu. Their bioconversion of flax lignan extract was investigated by resting cell catalysis and microbial fermentation, and the metabolism of SDG by Lactiplantibacillus plantarum C5 following fermentation was characterized by widely targeted metabolomics. Five L. plantarum strains producing ß-glucosidase with broad substrate specificity were isolated and identified, and they all can transform SDG into secoisolariciresinol (SECO). L. plantarum C5 resting cell reached a maximum SDG conversion of 49.19 ± 3.75%, and SECO generation of 21.49 ± 1.32% (0.215 ± 0.013 mm) at an SDG substrate concentration of 1 mM and 0.477 ± 0.003 mm SECO was produced at 4 mm within 24 h. Although sixteen flax lignan metabolites were identified following the fermentation of SDG extract by L. plantarum C5, among them, four were produced following the fermentation: SECO, demethyl-SECO, demethyl-dehydroxy-SECO and isolariciresinol. Moreover, seven lignans increased significantly. CONCLUSION: Fermentation significantly increased the profile and level of flax lignan metabolites, and the resting cell catalysis benefits from higher bioconversion efficiency and more straightforward product separation. Resting cell catalysis and microbial fermentation of flax lignan extract by the isolated ß-glucosidase production L. plantarum could be potentially applied in preparing flax lignan ingredients and fermented flaxseed. © 2024 Society of Chemical Industry.

The Impacts of Standardized Flaxseed Meal (XanFlax) on the Physicochemical, Textural, and Sensory Properties of Muffins.

Flaxseed is becoming increasingly popular as a superfood due to its many health benefits. While flaxseed is considered an oilseed, flaxseed meal (the by-product of flaxseed oil extraction) also contains many nutritional compounds not found in the oil. This study explored the use of a Canadian flaxseed (Linum usitatissimum L.) meal product to fortify bakery foods and improve their nutritional properties. Muffins were made using a control recipe as well as four different formulations that included varying amounts of a standardized flaxseed meal supplement called XanFlax (5, 10, 20, and 40%). The physicochemical properties of the muffins, including their texture, color, sugar content, pH, specific gravity, loss rate, and moisture, were evaluated. Additionally, the sensory attributes contributing to muffin quality were thoroughly examined. The lightness (L*) and yellowness (b*) of the muffins, which were highest in the control group at 82.22 and 34.69, respectively, decreased as the amount of XanFlax increased (p < 0.05). Additionally, the redness (a*) of the muffins increased as the amount of XanFlax increased (p < 0.05). The muffins' sugar content (2.00 brix%) remained consistent across all treatments and controls except for those prepared with 20% XanFlax (2.17 brix%). As the amount of XanFlax powder increased, the pH of the muffins increased significantly. The moisture content in the muffins was highest at 23.71 ± 0.79% in the 10% XanFlax treatment and lowest at 22.06 ± 0.30% in the 40% XanFlax treatment. The muffins enriched with 5% XanFlax had an average height of 5.35 cm and volume of 131.33 mL, surpassing the results for the muffins made with other formulas (p < 0.05). Additionally, the cohesiveness and gumminess of the muffins tended to increase with the addition of XanFlax. The most favorable attributes, namely the appearance, flavor, taste, texture, and overall acceptance, were consistently associated with the 5% and 10% XanFlax treatments (p < 0.05). This study marks the first time a standardized flaxseed gum product, XanFlax, has been described in a functional baking application.

Foaming with Starch: Exploring Faba Bean Aquafaba as a Green Alternative.

The demand for sustainable and functional plant-based products is on the rise. Plant proteins and polysaccharides often provide emulsification and stabilization properties to food and food ingredients. Recently, chickpea cooking water, also known as aquafaba, has gained popularity as a substitute for egg whites in sauces, food foams, and baked goods due to its foaming and emulsifying capacities. This study presents a modified eco-friendly process to obtain process water from faba beans and isolate and characterize the foam-inducing components. The isolated material exhibits similar functional properties, such as foaming capacity, to aquafaba obtained by cooking pulses. To isolate the foam-inducing component, the faba bean process water was mixed with anhydrous ethanol, and a precipitated fraction was obtained. The precipitate was easily dissolved, and solutions prepared with the alcohol precipitate retained the foaming capacity of the original extract. Enzymatic treatment with α-amylase or protease resulted in reduced foaming capacity, indicating that both protein and carbohydrates contribute to the foaming capacity. The dried precipitate was found to be 23% protein (consisting of vicilin, α-legumin, and ß-legumin) and 77% carbohydrate (amylose). Future investigations into the chemical structure of this foam-inducing agent can inform the development of foaming agents through synthetic or enzymatic routes. Overall, this study provides a potential alternative to aquafaba and highlights the importance of exploring plant-based sources for functional ingredients in the food industry.

Electrostatic field as an emergent technology in refining crude oils: a review.

Vegetable oils and fatty acid esters (FAEs) are commonly used in various industrial and commercial applications. However, the presence of contaminants in these oils can severely affect their functionality and suitability. Conventional refining techniques for vegetable oils typically involve degumming, neutralization, bleaching and deodorization. Meanwhile, refining of FAEs often utilize wet or dry washing processes. These are often resource-intensive, producing substantial waste products, causing neutral oil loss, and can also result in the loss of micronutrients. To address these challenges, researchers have explored the use of nano-adsorbents and electrostatic field (E-field) technologies as alternatives in purifying industrial dielectric oils by removing polar particles and contaminants. Nano-adsorbents demonstrated increased efficiency in removing polar contamination while minimizing neutral oil loss. However, removal of these spent adsorbents can be challenging due to their nano-size, and physicochemical properties. The use of these materials combined with E-field technologies offers a novel and sustainable solution for removing spent nano-adsorbents and contaminants. This review provides an overview of current traditional and novel refining technologies for vegetable oils and FAEs, including their associated limitations. Compared to conventional methods, E-field treatment offers several advantages, making it an attractive alternative to conventional approaches in food processing and oil refining.

A Metabolomics and Big Data Approach to Cannabis Authenticity (Authentomics).

With the increasing accessibility of cannabis (Cannabis sativa L., also known as marijuana and hemp), its products are being developed as extracts for both recreational and therapeutic use. This has led to increased scrutiny by regulatory bodies, who aim to understand and regulate the complex chemistry of these products to ensure their safety and efficacy. Regulators use targeted analyses to track the concentration of key bioactive metabolites and potentially harmful contaminants, such as metals and other impurities. However, the metabolic complexity of cannabis metabolic pathways requires a more comprehensive approach. A non-targeted metabolomic analysis of cannabis products is necessary to generate data that can be used to determine their authenticity and efficacy. An authentomics approach, which involves combining the non-targeted analysis of new samples with big data comparisons to authenticated historic datasets, provides a robust method for verifying the quality of cannabis products. To meet International Organization for Standardization (ISO) standards, it is necessary to implement the authentomics platform technology and build an integrated database of cannabis analytical results. This study is the first to review the topic of the authentomics of cannabis and its potential to meet ISO standards.

Availability of bioactive flax lignan from foods and supplements.

Hyperlipidemia, high levels of blood lipids including cholesterol and triglycerides, is a major risk factor for cardiovascular disease. Traditional treatments of hyperlipidemia often include lifestyle changes and pharmacotherapy. Recently, flaxseed has been approved as a nutrient that lowers blood lipids. Several metabolites of flaxseed lignan secoisolariciresinol diglucoside (SDG), have been identified that reduce blood lipids. SDG is present in flaxseed hull as an ester-linked copolymer with 3-hydroxy-3-methylglutaric acid (HMGA). However, purification processes involved in hydrolysis of the copolymer and enriching SDG are often expensive. The natural copolymer of SDG with HMGA (SDG polymer) is a source of bioactive compounds useful in prophylaxis of hypercholesterolemia. After consumption of the lignan copolymer, SDG and HMGA are released in the stomach and small intestines. SDG is metabolized to secoisolariciresinol, enterolactone and enterodiol, the bioactive forms of mammalian lignans. These metabolites are then distributed throughout the body where they accumulate in the liver, kidney, skin, other tissues, and organs. Successively, these metabolites reduce blood lipids including cholesterol, triglycerides, low density lipoprotein cholesterol, and lipid peroxidation products. In this review, the metabolism and efficacies of flaxseed-derived enriched SDG and SDG polymer will be discussed.

Depletion of cyanogenic glycosides in whole flaxseed via Lactobacillaceae fermentation.

Flaxseed is categorized as a functional food due to its abundance in oil, α-linolenic acid, dietary fibre, and lignan. However, flaxseed contains cyanogenic glycosides (CGs). Ingestion of CGs can influence nutrient absorption and induce adverse health effects. Due to the presence of CGs in flaxseed many countries prohibit the import and sale of flaxseed and flaxseed-based foods. In this study, whole flaxseed was fermented with a mixed culture of Lactobacillaceae (i.e., Lactobacillus sp., Limosilactobacillus sp., and Lactiplantibacillus sp.) and the concentration of CGs was determined. This process succeeded in completely removing CGs within 72 h in both bench-scale and scale-up studies. In addition, fatty acid composition in flaxseed remained unchanged and concentrations of flaxseed oil, and SDG in flaxseed were increased after fermentation. CG-free flaxseed products are beneficial, as they can be sold as health product ingredients, or as animal feed in markets that currently restrict the use of materials that contain CGs.

Uptake of Flaxseed Dietary Linusorbs Modulates Regulatory Genes Including Induction of Heat Shock Proteins and Apoptosis.

Flaxseed (Linum usitatissimum L.) is gaining popularity as a superfood due to its health-promoting properties. Mature flax grain includes an array of biologically active cyclic peptides or linusorbs (LOs, also known as cyclolinopeptides) that are synthesized from three or more ribosome-derived precursors. Two flaxseed orbitides, [1-9-NαC]-linusorb B3 and [1-9-NαC]-linusorb B2, suppress immunity, induce apoptosis in a cell line derived from human epithelial cancer cells (Calu-3), and inhibit T-cell proliferation, but the mechanism of LO action is unknown. LO-induced changes in gene expression in both nematode cultures and human cancer cell lines indicate that LOs promoted apoptosis. Specific evidence of LO bioactivity included: (1) distribution of LOs throughout the organism after flaxseed consumption; (2) induction of heat shock protein (HSP) 70A, an indicator of stress; (3) induction of apoptosis in Calu-3 cells; and (4) modulation of regulatory genes (determined by microarray analysis). In specific cancer cells, LOs induced apoptosis as well as HSPs in nematodes. The uptake of LOs from dietary sources indicates that these compounds might be suitable as delivery platforms for a variety of biologically active molecules for cancer therapy.

A review of flaxseed lignan and the extraction and refinement of secoisolariciresinol diglucoside.

Lignan is a class of diphenolic compounds that arise from the condensation of two phenylpropanoid moieties. Oilseed and cereal crops (e.g., flaxseed, sesame seed, wheat, barley, oats, rye, etc.) are major sources of plant lignan. Methods for commercial isolation of the lignan secoisolariciresinol diglucoside (SDG) are not well reported, as most publications describing the detection, extraction, and enrichment of SDG use methods that have not been optimized for commercial scale lignan recovery. Simply scaling up laboratory methods would require expensive infrastructure to achieve a marketable yield and reproducible product quality. Therefore, establishing standard protocols to produce SDG and its derivatives on an industrial scale is critical to decrease lignan cost and increase market opportunities. This review summarizes the human health benefits of flaxseed lignan consumption, lignan physicochemical properties, and mammalian lignan metabolism, and describes methods for detecting, extracting, and enriching flaxseed lignan. Refining and optimization of these methods could lead to the development of inexpensive lignan sources for application as an ingredient in medicines, dietary supplements, and other healthy ingredients.

Status of linusorbs in cold-pressed flaxseed oil during oxidation and their response toward antioxidants.

Linusorbs (LOs, cyclolinopeptides) are a class of naturally occurring cyclic hydrophobic peptides found in flaxseed oil, whose oxidation states indicate the oxidative stability and bitterness of flaxseed oil. Subjected to 63 °C accelerated oxidation, most Met-containing LOs in cold-pressed flaxseed oil entirely depleted by the 6th day except CLP, and MetO2-containing LOs became the dominant ones. However, no MetO2 form of Trp-containing LOs, such as CLD, CLF and CLG, were detected. Given their oxidative kinetics, methionine sulfoxide (MetO) residue in some LOs was less sensitive toward oxidation in the presence of Trp (Tryptophan) group, and the oxidative stability of Met-containing LOs was CLP < CLB < CLL ≈ CLM < CLO, as compared to MetO-containing LOs: CLD < CLE < CLC < CLF ≈ CLG. When antioxidant was added into cold-pressed flaxseed oil to assess the additives' antioxidant effect, no significant difference was observed on oil oxidative indices in early stage except α-tocopherol, where they vary dramatically in suppressing Met oxidation of LOs: L-AP (L-ascorbyl palmitate) > TBHQ (tert-butyl hydroquinone) > Î³-tocopherol > carnosic acid > α-tocopherol. Besides its ability to suppress oxidation of Trp-containing LOs, L-AP also exhibits superior antioxidant effect on non-Trp-containing LOs due to its amphiphilic property. Due to the prooxidative effects of both α- and γ-tocopherol on LOs that contain Trp, it has been suggested that tocopherols may repair Trp residue on LOs, leading to increased tendency of MetO residues to oxidize. The findings of this research are critical for elucidating the antioxidative mechanism of LOs, which can further lead to the establishment of strategies in suppressing bitter after taste to produce high-quality flaxseed oil.

Health benefits of flaxseed and its peptides (linusorbs).

Flaxseed (Linum usitatissimum L.) has been associated with numerous health benefits. The flax plant synthesizes an array of biologically active compounds including peptides or linusorbs (LOs, a.k.a., cyclolinopeptides), lignans, soluble dietary fiber and omega-3 fatty acids. The LOs arise from post-translational modification of four or more ribosome-derived precursors. These compounds exhibit an array of biological activities, including suppression of T-cell proliferation, excessive inflammation, and osteoclast replication as well as induction of apoptosis in some cancer cell lines. The mechanisms of LO action are only now being elucidated but these compounds might interact with other active compounds in flaxseed and contribute to biological activity attributed to other flax compounds. This review focuses on both the biological interaction of LOs with proteins and other molecules and comprehensive knowledge of LO pharmacological and biological properties. The physicochemical and nutraceutical properties of LOs, as well as the biological effects of certain LOs, and their underlying mechanisms of action, are reviewed. Finally, strategies for producing LOs by either peptide synthesis or recombinant organisms are presented. This review will be the first to describe LOs as a versatile scaffold for the action of compounds to deliver physiochemically/biologically active molecules for developing novel nutraceuticals and pharmaceuticals.

Mechanical stress on canola seed during dehulling.

The mature canola seed comprises an embryo, a group of structures that have a high content of protein and oil, and a fibrous seed coat held together by endosperm tissues. High-quality canola meal production requires removal of fiber-rich seed hull that typically contributes little nutrient value. Canola hull accounts for more than 70.0% of the insoluble fiber within the seed, which limits its uses as feed. Dehulling canola seed, before oil extraction, increases the protein content and decreases fiber content in the meal, to up to 50.0% and to down to 10.0%, respectively. Due to the complexity in dehulling canola seeds, several methods have been explored by the canola processing industry. However, little research has been conducted in analyzing the effects that mechanical dehulling methods have on seeds. In this study, we conducted mechanical tests to characterize seed behavior and hull fracture under uncoupled stresses such as compression and shear;stresses generated by the common dehulling equipment used in the industry, such as mills. All tests were carried out on single seeds at different moisture contents, from 5.00% to 25.0% wet mass basis (w.b.). Over a range of moisture contents studied, shear loading mode limited deformation of embryos in comparison to compression loading. Additionally, hardness tests were performed with a nano-indenter machine to determine seed hull properties. The overall results from these experiments provided information used to construct a computational model that represents canola seed deformation under compression stress.

The flax genome reveals orbitide diversity.

BACKGROUND: Ribosomally-synthesized cyclic peptides are widely found in plants and exhibit useful bioactivities for humans. The identification of cyclic peptide sequences and their precursor proteins is facilitated by the growing number of sequenced genomes. While previous research largely focused on the chemical diversity of these peptides across various species, there is little attention to a broader range of potential peptides that are not chemically identified. RESULTS: A pioneering study was initiated to explore the genetic diversity of linusorbs, a group of cyclic peptides uniquely occurring in cultivated flax (Linum usitatissimum). Phylogenetic analysis clustered the 5 known linusorb precursor proteins into two clades and one singleton. Preliminary tBLASTn search of the published flax genome using the whole protein sequence as query could only retrieve its homologues within the same clade. This limitation was overcome using a profile-based mining strategy. After genome reannotation, a hidden Markov Model (HMM)-based approach identified 58 repeats homologous to the linusorb-embedded repeats in 8 novel proteins, implying that they share common ancestry with the linusorb-embedded repeats. Subsequently, we developed a customized profile composed of a random linusorb-like domain (LLD) flanked by 5 conserved sites and used it for string search of the proteome, which extracted 281 LLD-containing repeats (LLDRs) in 25 proteins. Comparative analysis of different repeat categories suggested that the 5 conserved flanking sites among the non-homologous repeats have undergone convergent evolution driven by functional selection. CONCLUSIONS: The profile-based mining approach is suitable for analyzing repetitive sequences. The 25 LLDR proteins identified herein represent the potential diversity of cyclic peptides within the flax genome and lay a foundation for further studies on the functions and evolution of these protein tandem repeats.

Astragalin and Isoquercitrin Isolated from Aster scaber Suppress LPS-Induced Neuroinflammatory Responses in Microglia and Mice.

The current study investigated the anti-neuroinflammatory effects and mechanisms of astragalin (Ast) and isoquercitrin (Que) isolated from chamchwi (Aster scaber Thunb.) in the lipopolysaccharide (LPS)-activated microglia and hippocampus of LPS induced mice. LPS induced increased cytotoxicity, nitric oxide (NO) production, antioxidant activity, reactive oxygen species (ROS), inducible nitric oxide synthase (iNOS) expression, the release of pro-inflammatory cytokines, protein kinase B phosphorylation, and mitogen-activated protein kinases (MAPK) phosphorylation in LPS-treated microglial cells. Intraperitoneal injection of LPS also induced neuroinflammatory effects in the murine hippocampus. Ast and Que significantly reduced LPS-induced production of NO, iNOS, and pro-inflammatory cytokines in the microglia and hippocampus of mice. Therefore, anti-inflammatory effects on MAPK signaling pathways mediate microglial cell and hippocampus inflammation. In LPS-activated microglia and hippocampus of LPS-induced mice, Ast or Que inhibited MAPK kinase phosphorylation by extracellular signal-regulated kinase, c-Jun N-terminal kinase, and p38 signaling proteins. Ast and Que inhibited LPS-induced ROS generation in microglia and increased 1,1-diphenyl-2-picrylhydrazyl radical scavenging. In addition, LPS treatment increased the heme oxygenase-1 level, which was further elevated after Ast or Que treatments. Ast and Que exert anti-neuroinflammatory activity by down-regulation of MAPKs signaling pathways in LPS-activated microglia and hippocampus of mice.

Immunomodulatory Activity of Extracellular Vesicles of Kimchi-Derived Lactic Acid Bacteria (Leuconostoc mesenteroides, Latilactobacillus curvatus, and Lactiplantibacillus plantarum).

Lactic acid bacteria present in Kimchi, such as Leuconostoc mesenteroides (Lm), Latilactobacillus curvatus (Lc), and Lactiplantibacillus plantarum (Lp) produce extracellular vesicles (ECVs) that modulate immune responses. The ECVs of probiotic Kimchi bacteria are abbreviated as LmV, LcV, and LpV. Treatment of macrophages (RAW264.7) with ECVs (LmV, LcV, and LpV) increased the production of nitric oxide (NO), tumor necrosis factor (TNF)-α, and interleukin-6 (IL-6). Immunostimulatory effects exerted on the RAW264.7 cells were stronger after treatments with LmV and LcV than with LpV. Treatment of mice with LcV (1 mg/kg, orally) induced splenocyte proliferation and subsequent production of both NO and cytokines (INF-γ, TNF-α, IL-4, and IL-10). Furthermore, pre-treatment of macrophages and microglial cells with ECVs prior to LPS stimulation significantly attenuated the production of NO and pro-inflammatory cytokines (TNF-α, IL-1ß, and IL-6). Therefore, ECVs (LmV, LcV, and LpV) prevent inflammatory responses in the LPS-stimulated microglial cells by blocking the extracellular signal-regulated kinase (Erk) and p38 signaling pathways. These results showed that LmV, LcV, and LpV from Kimchi probiotic bacteria safely exert immunomodulatory effects.