Traditional Fermented Soybean Food Research Articles

Whole genome sequencing of the poly-γ-glutamic acid-producing novel Bacillus subtilis Tamang strain, isolated from spontaneously fermented kinema

Kinema, a traditional fermented soybean food from the Himalayas, is well-liked for its sticky texture and flavourful umami taste. Among 175 bacterial strains from spontaneously fermented kinema samples, Bacillus subtilis Tamang strain stood out for its high stickiness and viscosity. The strain’s Poly-γ-glutamic acid (γ-PGA) contains various groups of glutamic acid and has a molecular weight of 660 kDa. It demonstrates the ability to solubilize iron, preserve ferritin in Caco-2 cells, and exhibit antibacterial properties. The genome of B. subtilis Tamang is devoid of plasmid elements but does feature nine insert elements. Noteworthy is the presence of unique secondary metabolites with potential antimicrobial effects, such as amyloliquecidin GF610, bogorol A, and thermoactinoamide A. A total of 132 carbohydrate-active enzymes (CAZy) were identified, hinting at possible prebiotic characteristics. The genome analysis revealed genes responsible for γ-PGA production via the capBCA complex. Furthermore, genes associated with fibrinolytic activity, taste enhancement, biopeptides, immunomodulators, and vitamins like B12 and K2 were found, along with probiotics and various health benefits. The genetic material for L-asparaginase production, known for its anti-cancer properties, was also detected, as well as CRISPR-Cas systems. The absence of virulence factors and antimicrobial resistance genes confirms the safety of consuming B. subtilis Tamang as a food-grade bacterium.

Evaluation of Menopausal Syndrome Relief and Anti-Obesity Efficacy of the Korean Fermented Food Doenjang: A Randomized, Double-Blind Clinical Trial.

Foods that help improve menopausal syndrome are being studied worldwide. Doenjang is a traditional Korean fermented soybean food with potential health benefits for menopausal women. In this clinical trial using Doenjang, we aimed to compare the effectiveness of traditional Doenjang and commercial Doenjang in menopausal women. Furthermore, we compared whether Doenjang has a better effect if the number of beneficial microbes is higher. The analyses included the following groups: traditional Doenjang containing either a high dose (HDC; n = 18) or low dose (LDC; n = 18) of beneficial microbes and commercial Doenjang (CD; n = 20). The Kupperman index and hematological changes were examined before and after the use of Doenjang pills. The effects of Doenjang on obesity and body composition were studied before and after ingestion. Lastly, the microorganisms and short-chain fatty acid changes in the stool were compared. The Kupperman index decreased after Doenjang consumption in all three groups, with the greatest decrease in the LDC group. Only the groups that took traditional Doenjang pills exhibited reduced LDL cholesterol. No changes in obesity and inflammation-related indicators were observed. The number of Firmicutes, associated with obesity, decreased in the CD group but the numbers of Bacteroidetes increased in the HDC and CD groups. Thus, traditional Doenjang is more effective in alleviating menopausal syndrome than commercial Doenjang. Further research on the anti-obesity effect or changes in microbiomes and short-chain fatty acids in feces is needed.

Correlation between monosaccharide, oligosaccharide, and microbial community profile changes in traditional soybean brick (meju) fermentation

Meju is essential for making diverse traditional fermented soybean foods in Korea. To understand the changes in carbohydrates during fermentation, we aimed to identify autochthonous microorganisms from spontaneously fermented meju and compare the alterations in monosaccharides and oligosaccharides throughout the fermentation process. Microbial diversity was determined using a metabarcoding approach, and monosaccharide and oligosaccharide profiles were obtained by HPLC-Q-TOF MS and HPLC-MS/MS analyses, respectively. The dominant bacterial genera were Weissella, Lactobacillus, and Leuconostoc, while Mucor was highly abundant in the fungal community. The total monosaccharide content increased from Day 0 to Day 50, with the highest amount being 4.37 mg/g. Oligosaccharide profiling revealed the degradation of soybean dietary fiber during fermentation, and novel oligosaccharide structures were also discovered. Correlation analysis revealed that the fungus Mucor was positively related to pentose-containing oligosaccharides, galactose, and galacturonic acid, indicating that Mucor may degrade soybean dietary fibers such as xylogalacturonan, arabinogalactan, and rhamnogalacturonan. The negative relationships between the abundances of Weissella and oligo- and monosaccharides suggested that the bacteria may utilize saccharides for fermentation. These findings provide insights into the mechanisms underlying carbohydrate degradation and utilization; the key components involved in saccharide transformation that contribute to the characteristics of traditional meju were subsequently identified.

Degradation of Bisphenol A by Bacillus subtilis P74 Isolated from Traditional Fermented Soybean Foods.

Bisphenol A (BPA), one of the most widely used plasticizers, is an endocrine-disrupting chemical that is released from plastic products. The aim of this study was to screen and characterize bacteria with excellent BPA-degrading abilities for application in foods. BPA degradation ability was confirmed in 127 of 129 bacterial strains that were isolated from fermented soybean foods. Among the strains, B. subtilis P74, which showed the highest BPA degradation performance, degraded 97.2% of 10 mg/L of BPA within 9 h. This strain not only showed a fairly stable degradation performance (min > 88.2%) over a wide range of temperatures (30-45 °C) and pH (5.0-9.0) but also exhibited a degradation of 63% against high concentrations of BPA (80 mg/L). The metabolites generated during the degradation were analyzed using high-performance liquid chromatography-mass spectrometry, and predicted degradation pathways are tentatively proposed. Finally, the application of this strain to soybean fermentation was conducted to confirm its applicability in food.

A COMPUTATIONAL GENOME ANALYSIS OF STRAIN BACILLUS SUBTILIS MIZ-8 ISOLATED FROM BEKANG REVEALS A DISTINCT CHROMOSOME AND PLASMID CONFERRING SELECTIVE ADVANTAGE

Bacillus subtilis Miz-8 is isolated from Bekang, a traditional fermented soybean food of Mizoram, India. Here, we report computationally analyzed strain Miz-8 mapped to the annotated genome of the reference strain Bacillus subtilis subsp. natto BEST195. Miz-8 has a circular chromosome and a small plasmid with genome size of 4105264 bp and 5838 bp, respectively. Genome contains the genes responsible for synthesis of biosynthetic metabolites like surfactin, fengycin, bacillibactin, bacilysin, subtilosin A and alpha amylase production. The strain Miz-8 harbored virulence genes identical to the strain BEST195 render themselves harmless for human consumption. However, Strain Miz-8 has one intact prophage region but no integrase protein, making it incapable of lateral transfer of antimicrobials. Antibiotic resistance genes were predicted among which tmrB gene was on perfect hit. Plasmid of strain Miz-8 contains no prophage sequences and antibiotic resistance genes. Furthermore, there were several single nucleotide polymorphisms and 344 insertion-deletion polymorphisms. Bacillus subtilis Miz-8's genomic information unmasks its functional significance on human health.

Bitter Peptides in Fermented Soybean Foods - A Review.

Fermented soybean foods with a long history are popular worldwide because of rich nutrition. However, many traditional fermented soybean foods have unacceptable bitterness, which mostly comes from the bitter peptides produced from the hydrolysis of soybean proteins. In this review, the bitter peptides in fermented soybean foods is briefly reviewed. The structural properties of bitter receptors and bitter peptides were reviewed. Bitterness is perceived through the binding between bitter compounds and specific sites of bitter receptors (25 hTAS2Rs), which further activate the downstream signal pathway mediated by G-protein. And it converts chemical signals into electrical signals, and transmit them to the brain. In addition, the influencing factors of bitter peptides in fermented soybean foods were summarized. The bitterness of fermented soybean foods primarily results from the raw materials, microbial metabolism during fermentation, unique techniques, and interactions of various flavor compounds. Moreover, the structure-bitterness relationship of bitter peptides was also discussed in this review. The bitterness degree of the bitter peptide is related to the polypeptide hydrophobicity, amino acids in the peptide, peptide molecular weight and polypeptide spatial structure. Studying the bitter peptides and their bitter characteristics in fermented soybean foods is beneficial for improving the sensory quality of fermented soybean foods and prompting more consumers accept them.

Evaluation of Body Changes and the Anti-Obesity Effect after Consumption of Korean Fermented Food, Cheonggukjang: Randomized, Double-Blind Clinical Trial.

Cheonggukjang is a traditional Korean fermented soybean food with potential health benefits. For this reason, Cheonggukjang is consumed in the form of pills in addition to being used as a food ingredient. There are few clinical studies that have evaluated changes in various health indicators through blood and stool tests before and after consumption of Cheonggukjang. In this study, symptoms and hematological changes were analyzed before and after the intake of traditional Cheonggukjang pills containing high-dose (n = 19) or low-dose (n = 20) beneficial bacteria and commercial Cheonggukjang pills (n = 20). Anti-obesity effects and body composition changes were determined before and after Cheonggukjang consumption. Lastly, the changes in microorganisms and short-chain fatty acids in the stool were compared. No changes in obesity and inflammation-related indicators were observed before and after Cheonggukjang consumption. The Firmicutes/Bacteroidetes ratio, associated with obesity, decreased in all three groups after Cheonggukjang consumption, but no statistical significance was indicated. Cheonggukjang contained various BAs, but they did not adversely affect symptoms and hematological changes in the participants. BAs generated during the manufacturing process of Cheonggukjang did not have any adverse effects in this randomized, double-blind clinical trial. Further research is needed in future concerning the anti-obesity effect or regarding changes in the microbiome and short-chain fatty acids in feces.

Microbiome analysis of thai traditional fermented soybeans reveals short-chain fatty acid-associated bacterial taxa

Thua Nao is a Thai traditional fermented soybean food and low-cost protein supplement. This study aimed to evaluate the bacterial community in Thua Nao from northern Thailand and assess potentially active short-chain fatty acids (SCFAs)-related bacteria. Sixty-five Thua Nao consisting of 30 wet and 35 dried samples were collected from six provinces: Chiang Rai, Chiang Mai, Mae Hong Son, Lampang, Lamphun, and Phayao. Bacterial diversity was significantly higher in the wet samples than in the dried samples. The dominant phyla were Firmicutes (92.7%), Proteobacteria (6.7%), Actinobacteriota (0.42%), and Bacteroidota (0.26%). The genus Bacillus (67%) was the most represented in all samples. Lactobacillus, Enterococcus, and Globicatella were enriched in the wet samples. Assessment of the SCFA-microbiota relationships revealed that high butyrate and propionate concentrations were associated with an increased Clostridiales abundance, and high acetate concentrations were associated with an increased Weissella abundance. Wet products contained more SCFAs, including acetate (P = 2.8e−08), propionate (P = 0.0044), butyrate (P = 0.0021), and isovalerate (P = 0.017), than the dried products. These results provide insight into SCFA-microbiota associations in Thua Nao, which may enable the development of starter cultures for SCFA-enriched Thua Nao production.

Antibacterial potential of an ethnic probiotic containing food, Akhuni/Axone and its chemical characterization

‘Akhuni/Axone’, a popular traditional fermented soybean food of northeast India, was evaluated for antibacterial activity against Staphylococcus aureus and Escherichia coli along with its mechanism of action and chemical characterization. The effects of “Akhuni/Axone” treatment on these bacteria were assessed using the following methods: zone of inhibition, minimum inhibitory concentration (MIC), minimum bactericidal concentrations (MBC), time kill curve analysis, cellular leakage measurement, oxidative stress analysis, EtBr influx/efflux assay and SEM microscopy. Among petroleum ether, ethyl acetate, ethanol and methanol extracts of ‘Akhuni/Axone’, ethyl acetate extract (AKEA) showed significant antibacterial activity against both the pathogens. The MIC of AKEA was found to be 8.09 ± 0.86 mg/ml and 10.79 ± 0.91 mg/ml against S. aureus and E. coli respectively whereas MBC values were 10.79±0.63 mg/ml and 13.45 ± 1.12 mg/ml against S. aureus and E. coli respectively. The time-kill analysis and cellular leakage assay results indicated that AKEA has bactericidal effect in a concentration and time dependent manner towards both the pathogens. Oxidative stress analysis indicated that AKEA treatment led to excessive ROS production and oxidative stress. EtBr influx/efflux assay indicated AKEA as efflux pump inhibitor and SEM microscopy confirmed the morphological changes on the cell wall and cytoplasmic membrane of bacteria. The chemical analysis of AKEA through 1H NMR, 13C NMR and GC-MS showed a high abundance of triglycerides. These findings indicate that ‘Akhuni/Axone’ can inhibit bacteria, S aureus and E coli and results from this study will be helpful in further studies on other antimicrobial compounds and for developing food-based natural preservatives.

Glucoregulatory Properties of Fermented Soybean Products

Type 2 diabetes mellitus is a chronic metabolic disease, characterized by persistent hyperglycemia, the prevalence of which is on the rise worldwide. Fermented soybean products (FSP) are rich in diverse functional ingredients which have been shown to exhibit therapeutic properties in alleviating hyperglycemia. This review summarizes the hypoglycemic actions of FSP from the perspective of different target-related molecular signaling mechanisms in vitro, in vivo and clinical trials. FSP can ameliorate glucose metabolism disorder by functioning as carbohydrate digestive enzyme inhibitors, facilitating glucose transporter 4 translocation, accelerating muscular glucose utilization, inhibiting hepatic gluconeogenesis, ameliorating pancreatic dysfunction, relieving adipose tissue inflammation, and improving gut microbiota disorder. Sufficiently recognizing and exploiting the hypoglycemic activity of traditional fermented soybean foods could provide a new strategy in the development of the food fermentation industry.

α-Glucosidase Inhibitory Activity of Fermented Okara Broth Started with the Strain Bacillus amyloliquefaciens SY07.

In this work, a new strain of Bacillus amyloliquefaciens SY07 isolated from a traditional fermented soybean food was reported to possess remarkable α-glucosidase inhibitor-producing ability. Different culture media were applied for the proliferation of B. amyloliquefaciens SY07, and it was found that fermented okara broth presented the highest α-glucosidase inhibitory activity, while Luria-Bertani medium showed a negative effect. The extract from fermented okara broth acted in a dose-dependent manner to inhibit α-glucosidase activity, with an IC50 value of 0.454 mg/mL, and main inhibitors in the fermentation extract presented a reversible, uncompetitive pattern according to Lineweaver–Burk plots. Moreover, 1-deoxynojirimycin, a recognized α-glucosidase inhibitor, was found in the extract. Results indicated that B. amyloliquefaciens SY07 could utilize okara, a by-product from the soy processing industry, to generate α-glucosidase inhibitors effectively, and be regarded as a novel excellent microbial candidate for safe, economical production of potential functional foods or ingredients with hypoglycemic effect.

Bacillus subtilis BN strain promotes Th1 response via Toll-like receptor 2 in polarized mouse M1 macrophage.

Bacillus subtilis BN strain (BN strain) was isolated from natto, a traditional Japanese fermented soybean food product. The present study investigated the Th1 responses of the BN strain on a mouse macrophage cell line, J774.1. In cell cultures, the BN strain (spore cell cultured in Schaeffer's sporulation media) significantly increased the production of interleukin (IL-)12 protein. The BN strain induced the mRNA expression of M1 polarization genes, such as inducible nitric oxide synthase and IL-12p40 mRNA, and suppressed the mRNA expression of intracellular marker genes of M2 polarization, such as arginase 1 mRNA. The BN strain downregulated the mRNA expression of Toll-like receptor 4 (TLR4), while it upregulated the mRNA expression of TLR2, MyD88, and nuclear factor kappa B (NF-κB). The production of IL-12 protein induced by the BN strain was decreased by inhibitors of MyD88, NF-κB, and IκB kinase. Moreover, the production of IL-12 was strongly suppressed by neutralizing antibody against TLR2. These results suggest that the BN strain promotes Th1 response via TLR2 signal in mouse M1 macrophage. PRACTICAL APPLICATIONS: Bacillus subtilis is known to have beneficial effects for the host. B. subtilis BN stain (BN strain) was isolated from natto, a traditional Japanese fermented soybean food product. The effects of the BN strain on the Th1 response in macrophage cell cultures were investigated in this work. We found that the spore cells of BN strain promoted the production of Th1-type cytokine, and induced macrophage M1 polarization via Toll-like receptor 2. This study can serve as a significant reference for the development of functional food and feed with immunostimulatory effects. Over time, new food and feed products containing the BN strain may emerge, such as Juice, powder, and tablet.

Protective effect of traditional Korean fermented soybean foods (doenjang) on a dextran sulfate sodium-induced colitis mouse model.

Objective: The cause of ulcerative colitis (UC) is unknown, and the use of anti-inflammatory and immunosuppressive drugs with certain side effects is currently replacing treatment. Therefore, it is important to find new healthy foods or ingredients that exhibit potential protective and anti-inflammatory effects on UC. This study investigated the potential protective effect of doenjang on dextran sulfate sodium (DSS)-induced colitis in a mouse model. Materials and methods: Four doenjang samples (TCD21-51-1, TCD21-55-1, TMD21-16-1, and TFD21-1-1) were used. To examine the effects of the four doenjang samples on UC caused by DSS in a mouse model, the clinical symptoms of UC, such as body weight, disease activity index (DAI), and colon macroscopic damage index (CMDI) were analyzed. Moreover, immune-related blood cell counts, serum levels and protein expression of tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6), and nitric oxide (NO) production were measured in DSS-induced UC in mice for analysis. Results: The four doenjang samples increased the colon length shortened by DSS, reduced DAI (diarrhea and hemoccult), CMDI (ulceration, inflammation, and hemorrhage) and the content of immune-related cells in the blood. Moreover, the levels of TNF-α, IL-6, and NO increased by DSS were decreased by doenjang, and tissue damage was significantly reduced. Conclusions: These findings confirmed that doenjang exerts protective effects against UC, suggesting its possible use in developing therapeutic strategies or functional products.

Contamination characteristics and risk assessment of aflatoxins in homemade soybean paste, a traditional fermented soybean food, in South Korea

Soybean paste is manufactured through microbial fermentation and may become contaminated with aflatoxins. Herein, we conducted nationwide large-scale monitoring (n = 1436) over three years (2018–2020) to investigate aflatoxin levels according to geographic, demographic, manufacturing, quality factors, and risk characteristics of homemade soybean paste produced through fermentation. The mean level of total aflatoxins was 5.88 μg/kg (range, 0.01–281.92), with the most common contaminating type being the B type. Aflatoxin levels significantly differed according to the region, age of the manufacturer, type of starter used, and the amino-type nitrogen content and pH of the homemade soybean paste (p < 0.05). Aflatoxin levels was significantly higher when starters were manufactured using the traditional method (inoculation with a naturally occurring strain in the surrounding environment). The aflatoxin exposure level estimated through the average intake of homemade soybean paste in all age groups was 0.1012 ng/kg body weight/day. The risk assessment for the genotoxic and carcinogenic potential of aflatoxins using the margin of exposure approach revealed values of 3705–3954 for average intake of homemade soybean paste, indicating public health concern. These results suggest that follow-up studies and safety management strategies are needed to reduce aflatoxin levels in homemade soybean paste.

Natto extract, a Japanese fermented soybean food, directly inhibits viral infections including SARS-CoV-2 in vitro

Natto, a traditional Japanese fermented soybean food, is well known to be nutritious and beneficial for health. In this study, we examined whether natto impairs infection by viruses, such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) as well as bovine herpesvirus 1 (BHV-1). Interestingly, our results show that both SARS-CoV-2 and BHV-1 treated with a natto extract were fully inhibited infection to the cells. We also found that the glycoprotein D of BHV-1 was shown to be degraded by Western blot analysis and that a recombinant SARS-CoV-2 receptor-binding domain (RBD) was proteolytically degraded when incubated with the natto extract. In addition, RBD protein carrying a point mutation (UK variant N501Y) was also degraded by the natto extract. When the natto extract was heated at 100 °C for 10 min, the ability of both SARS-CoV-2 and BHV-1 to infect to the cells was restored. Consistent with the results of the heat inactivation, a serine protease inhibitor inhibited anti-BHV-1 activity caused by the natto extract. Thus, our findings provide the first evidence that the natto extract contains a protease(s) that inhibits viral infection through the proteolysis of the viral proteins.

Comparison of volatile and non-volatile metabolites in sufu produced with bacillus licheniformis by rapid fermentation

ABSTRACT Sufu is a pleasant-tasting, traditional Chinese fermented soybean food that is rich in nutrients. In this study, the changes of volatile and nonvolatile metabolites in sufu fermented by bacillus licheniformis, were investigated. The results indicated that a total of 55 kinds of nonvolatile compounds were detected, including 2 carbohydrates, 4 alcohols, 17 amino acids, 18 organic acids, 6 biogenic amines, and 8 other substances. Furthermore, a total of 58 volatile compounds identified were composed of 11 esters, 16 alcohols, 10 acids, and 21 miscellaneous compounds. Inoculation of bacillus licheniformis enriched the metabolite profile of sufu and improved its functionality and safety of edibility. It was observed that the pure fermented starter resulted in controlled acceleration of sufu maturation.

Metagenomic analysis of bacterial community structure and functions during the fermentation of da-jiang, a Chinese traditional fermented food

The objective of this study was to understand the role of bacteria during the fermentation of da-jiang, a popular traditional Chinese fermented soybean food. In this study, 15 homemade da-jiang samples were analyzed bacteria community via high-throughput sequencing technology. The sequencing data analysis showed that 248 genera, including 841 species of bacteria, the dominant bacteria were Leuconostoc, Acinetobacter, Enterobacter, Enterococcus, Pantoea, Lactococcus, Lactobacillus, Pseudomonas, Staphylococcus, Tetragenococcus,and Weissella. Furthermore, the functional genes of bacteria during the fermentation are mainly associated with carbohydrate metabolism, amino acid metabolism, membrane transport, inorganic ion transport and metabolism, signal transduction,and metabolism of coenzyme and vitamins. Most the bacteria can participate in primary metabolism, the characteristics of da-jiang related to a few species. Pseudomonas Chlororaphis is closely associated with the color of da-jiang. Pseudomonas fluorescens is closely associated to the production of tyramine and histamine. This information may provide important insight into the microbial community structure and key enzymes in traditional da-jiang and might help elucidate the biological functions of these microbes, as well as their use as a starter culture to obtain high-quality da-jiang.

High-Throughput Sequencing and Metabolomics Reveal Differences in Bacterial Diversity and Metabolites Between Red and White Sufu.

Sufu is a traditional fermented soybean food produced in China. However, the microbial compositions and metabolites of different types of sufu have not been studied in detail. Accordingly, in this study, we evaluated the differences in bacterial communities and metabolites between commercial red sufu (RS) and white sufu (WS). Principal coordinate analysis and the unweighted pair group method with arithmetic means analysis of 16S rRNA genes revealed that the bacterial community structures of RS and WS differed dramatically. At the phylum level, the relative abundances of Firmicutes and Proteobacteria were significantly different between RS and WS (P < 0.01). Moreover, the abundances of Lactococcus and Tetragenococcus genera were significantly different between RS and WS (P < 0.01). Among metabolites, most free amino acids, few of volatile flavor compounds, and some organic acids showed significant differences between RS and WS (P < 0.05). Additionally, correlations between microbiota and metabolites were determined. Aggregated boosted tree analysis showed that formic acid had the highest relative influence (20.27%) on bacterial community diversity (Chao 1), following by arginine (5.38%), propanol (4.57%), oxalic acid (4.46%), and hexanol (4.43%). Moreover, Streptococcaceae and Moraxellaceae had the highest relative influence on the concentration of formic acid (12.84% and 8.75%, respectively). The profiles obtained in this study improve our understanding of the relationships between bacterial flora and metabolites in different types of sufu. These findings may help us interpret the roles of bacterial communities in the flavor and characteristics of sufu.

Chungkookjang, a soy food, fermented with Bacillus amyloliquefaciens protects gerbils against ishcmeic stroke injury, and post-stroke hyperglycemia.

Chungkookjang is a traditional Korean fermented soybean food with anti-diabetic and thrombolytic activity. It may also have anti-stroke activity. We determined that chungkookjang made with Sunchang Research Center for Fermentation Microbes 100730 and 100731 strains of Bacillus amyloliquefaciens(SRCM100730; CKJ730) and SRCM100731(CKJ731) protected against ischemic stroke and post-stroke hyperglycemia in Mongolian gerbils with ischemic stroke induced by transient occlusion of the carotid arteries. Gerbil fed 4.5% cooked soybeans (CSB), CKJ730, CKJ731, or cellulose (negative-control) in 40 energy% fat diets for 3 weeks, and then had artery occlusion for 8 min and continued taking the assigned diet for 5 weeks. CKJ730 and CKJ731 had thrombolytic activity and prevented neuronal cell death. Consequently, they improved short-term memory and spontaneous alteration compared to the negative-control. CKJ730 and CKJ731 improved neurological symptoms including drooped eyes, crouched posture, flexor reflex, and walking patterns the most among the stroke-induced gerbils. CKJ730 and CKJ731 increased active time, grip strength, and blood flow measured by Doppler compared to the negative-control. CKJ730 protected against post-stroke glucose dysregulation by restoring β-cell mass in the gerbils with transient artery occlusion. Serum tumor necrosis factor-α and interleukin-1β levels were lower in CKJ730 and CKJ731 than the negative-control. CSB also improved glucose metabolism and suppressed inflammatory cytokines, but less than CKJ730 and CKJ731. Clostridia increased, and Bacteriodia slightly decreased in the negative-control group, compared to the normal-control. CKJ730 and CKJ731 changed the amounts of Bacteriodia and Clostridia to be similar to normal-control. In conclusion, the daily intake of chungkookjang fermented with B. amyloliquefaciens improved the gut microbiome, increased blood flow to the brain, suppressed systemic inflammation, and may reduce the susceptibility to injury from ischemic stroke in gerbils subjected to ischemic injury.

High-throughput sequencing approach to characterize dynamic changes of the fungal and bacterial communities during the production of sufu, a traditional Chinese fermented soybean food

Red sufu is a traditional food produced by the fermentation of soybean. In this study, sufu samples were periodically collected during the whole fermentation to investigate the dynamic changes of fungal and bacterial communities using high-throughput sequencing technology. The overall process can be divided into pre- and post-fermentation. During post-fermentation, the pH value showed a gradual decrease over time while the amino nitrogen content increased. Trichosporon, Actinomucor and Cryptococcus were the main genera in pre-fermentation while Monascus and Aspergillus were dominant in post-fermentation. This huge shift in fungal composition was caused by process procedure of pouring dressing mixture. However, the bacterial composition was not greatly changed after pouring dressing mixture, the Acinetobacter and Enterobacter were the predominant genera throughout the whole process. Furthermore, Bacillus species were first detected after adding dressing mixture, but declined abruptly to a very low level (0.07%) by the end of the fermentation. Our work demonstrates the dynamic changes of physicochemical properties and microbial composition in every fermentation stage, the knowledge of which could potentially serve as a foundation for improving the safety and quality of sufu in the future.