Whole-genome analysis, evaluation and regulation of in vitro and in vivo GABA production from Levilactobacillus brevis YSJ3.

Gamma-aminobutyric acid (GABA) produced by lactic acid bacteria (LAB) is safe and has several health benefits. Levilactobacillus brevis YSJ3 was selected from 110 LAB. It exhibited the highest in vitro GABA production level of 970.10 µg/mL. Whole-genome analysis revealed that L. brevis YSJ3 contained gadR, gadC, gadB and gadA. Furthermore, the Luedeking-Piret model was fitted, which indicated that GABA production was divided into three stages. The gadR 0079, gadC 0080, and gadB 0081 were confirmed to promote GABA synthesis. Moreover, 55 metabolites, particularly those involved in arginine metabolism, were significantly different at 6 and 20 h of cultivation. Notably, L. brevis YSJ3 significantly improved sleep in mice and increased GABA levels in the mice's gut compared with the control group. This suggests that the oral administration of L. brevis YSJ3 improves sleep quality, probably by increasing intestinal GABA levels. Overall, L. brevis YSJ3 was confirmed as a GABA-producing strain in vitro and in vivo, making it a promising probiotic candidate for its application in food and medicine.

Variation in glucosinolates and the formation of functional degradation products in two Brassica species during spontaneous fermentation.

Vegetables from the Brassica species are excellent sources of glucosinolates (GLSs), the precursors of health-promoting isothiocyanates (ITCs). Fermentation enhances the biotransformation of GLSs into potential bioactive ITCs. To explore the biotransformation of GLSs during Brassica fermentation, the changes in GLSs during the fermentation of two Brassica species (i.e., cauliflower and broccoli); the formation of corresponding breakdown products; and the shifts in physicochemical parameters, bacterial communities, and myrosinase activities involved in GLSs degradation were systematically investigated. Nine aliphatic, three indolic, and two benzenic GLSs were identified in fermented cauliflower (FC) and fermented broccoli (FB). Aliphatic glucoiberin and glucoraphanin were the major forms of GLS in FC and FB, respectively; indolic glucobrassicin was also abundant in both FC and FB. The total GLS content decreased by 85.29% and 65.48% after 3 d of fermentation in FC and FB, respectively. After 2 d of fermentation, a significant increase in bioactive GLS degradation products (P < 0.05), including sulforaphane (SFN), iberin (IBN), 3,3-diindolylmethane (DIM), and ascorbigen (ARG), was observed in FC and FB compared to in fresh cauliflower and broccoli. Moreover, variations in pH value and titratable acidity in FC and FB correlated with Brassica fermentation and were accomplished by lactic acid bacteria, including Weissella, Lactobacillus-related genera, Leuconostoc, Lactococcus, and Streptococcus. These changes may enhance the biotransformation of GSLs to ITCs. Overall, our results indicate fermentation leads to the degradation of GLSs and the accumulation of functional degradation products in FC and FB.

Reduced formation of biogenic amines in low-salt Zhacai via fermentation under CO2-modified atmosphere.

Reducing sodium salt content in traditional fermented vegetables and developing low-salt fermented products have attracted increasing attention.However, low-salt fermented vegetables are prone to accumulate toxic biogenic amines (BAs) caused by the undesirable metabolism of spoilage microorganisms. This study aimed to investigate the impact of a CO2-modified atmosphere (MA) approach to the fermentation of low-salt Zhacai and the accumulation of BAs. The results show CO2-MA effectively suppressed the production of excessive BAs in low-salt Zhacai, as evidenced by a decrease in the total BA content from 63.66 to 161.41 mg/ kg under natural air conditions to 1.88-24.76 mg/ kg under CO2-MA. Overall, the mechanism of hindering BA formation was closely related to the change in the microbial community and the downregulation of BA-producing enzymes. Lactic acid bacteria, including Lactiplantibacillus plantarum, Weissella spp., and Pediococcus spp., were enriched under CO2-MA, whereas amine-producing microorganisms (e.g., Halomonas spp., Psychrobacter spp., Corynebacterium spp., and Levilactobacillus brevis) were greatly inhibited. Moreover, metagenomic analysis revealed that genes encoding amino acid decarboxylase, amine deiminase, and amine synthase were downregulated, which could be the fundamental reason for BA reduction. This study provides an alternative method for reducing BA production in fermented food.

Dynamics of the glucosinolate-myrosinase system in tuber mustard (Brassica juncea var. tumida) during pickling and its relationship with bacterial communities and fermentation characteristics.

Pickled tuber mustard is a traditional fermented pickle widely consumed in China, and it is characterized by the presence of glucosinolates (GSLs). To understand the biotransformation of GSLs in tuber mustard during pickling, the dynamics of the glucosinolate-myrosinase (GSL-MYR) system and its potential associations with bacterial communities and fermentation characteristics (i.e., salinity, titratable acidity [TAA], and pH) were investigated. In total, 18 GSLs were identified in fresh tuber mustard; 12 were aliphatic, 4 were indolic, and 2 were aromatic, with aliphatic sinigrin and aromatic gluconasturtiin being the dominating components. The pickling process resulted in complete degradation of GSLs, with isothiocyanates (ITCs) and nitriles being the main breakdown products. Total ITCs reached maximum concentrations on day 21-28, while total nitriles peaked at the end of pickling. Based on Spearman's correlation analysis, our study showed that lactic acid bacteria (LAB) species might contribute to GSL transformation in pickled tuber mustard. Specifically, Weissella paramesenteroides, Pediococcus pentosaceus, and unclassified Lactococcus exhibited positive correlations with GSL contents (p < 0.01), suggesting that they might contribute to the increasing amounts of GSLs in the initial pickling, while the Lactobacillus-related populations that dominated in the later stages (i.e., Companilactobacillus alimentarius and Lactiplantibacillus Plantarum) were positively correlated with nitrile product concentrations. Moreover, redundancy analysis showed that pH and TAA had strong effect on myrosinase activity during tuber mustard pickling, which was dictated via the organic acids produced by microorganisms. This study provided a perspective for understanding the effect of fermentation on the transformation of tuber mustard GSLs.

Changes in Phytochemical Profiles and Biological Activity of Olive Leaves Treated by Two Drying Methods.

Olive leaves, which are the most abundant byproducts of the olive industry, offer multiple health benefits. The investigation of the phytochemical profiles and relevant biological activities is an essential step toward transforming these low-value byproducts into value-added ones. This study systematically investigated the phytochemical profiles, antioxidant capacity, and inhibition rates of olive leaves from four cultivars on the α-glucosidase, α-amylase, and angiotensin-converting enzyme (ACE). The leaves were prepared using two common drying methods, namely, hot air-drying and freeze-drying. A total of 33 bioactive compounds were identified in the olive leaves, namely, 19 flavonoids, 2 phenylethanoids, 2 coumarins, 2 hydroxycinnamic acids, 2 iridoids, and 6 triterpenic acids. Quantification of the bioactive compounds revealed high amounts of polyphenols, especially flavonoids [2,027-8,055 mg/kg dry weight (DW)], iridoids (566-22,096 mg/kg DW), and triterpenic acids (13,824-19,056 mg/kg DW) in the olive leaves. The hot air-dried leaves showed significantly (P < 0.05) higher iridoid (oleuropein and secoxyloganin) content than the fresh leaves, while freeze-drying resulted in significantly (P < 0.05) higher flavonoid aglycone and hydroxytyrosol content. Additionally, freeze-drying led to samples with the highest radical scavenging, α-amylase, α-glucosidase, and ACE inhibition abilities. The flavonoid (e.g., quercetin, luteolin, eriodictyol, kaempferol-7-O-glucoside, and luteolin-7-O-glucoside), hydroxytyrosol, and oleanolic acid contents in the olive leaves were positively correlated (P < 0.05) with their bioactive potentials.

Comparative Evaluation of the Phytochemical Profiles and Antioxidant Potentials of Olive Leaves from 32 Cultivars Grown in China.

Olives (Olea europaea L.) are a significant part of the agroindustry in China. Olive leaves, the most abundant by-products of the olive and olive oil industry, contain bioactive compounds that are beneficial to human health. The purpose of this study was to evaluate the phytochemical profiles and antioxidant capacities of olive leaves from 32 cultivars grown in China. A total of 32 phytochemical compounds were identified using high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry, including 17 flavonoids, five iridoids, two hydroxycinnamic acids, six triterpenic acids, one simple phenol, and one coumarin. Specifically, olive leaves were found to be excellent sources of flavonoids (4.92-18.29 mg/g dw), iridoids (5.75-33.73 mg/g dw), and triterpenic acids (15.72-35.75 mg/g dw), and considerable variations in phytochemical content were detected among the different cultivars. All tested cultivars were classified into three categories according to their oil contents for further comparative phytochemicals assessment. Principal component analysis indicated that the investigated olive cultivars could be distinguished based upon their phytochemical profiles and antioxidant capacities. The olive leaves obtained from the low-oil-content (<16%) cultivars exhibited higher levels of glycosylated flavonoids and iridoids, while those obtained from high-oil-content (>20%) cultivars contained mainly triterpenic acids in their compositions. Correspondingly, the low-oil-content cultivars (OL3, Frantoio selection and OL14, Huaou 5) exhibited the highest ABTS antioxidant activities (758.01 ± 16.54 and 710.64 ± 14.58 mg TE/g dw, respectively), and OL9 (Olea europaea subsp. Cuspidata isolate Yunnan) and OL3 exhibited the highest ferric reducing/antioxidant power assay values (1228.29 ± 23.95 mg TE/g dw and 1099.99 ± 14.30 mg TE/g dw, respectively). The results from this study may be beneficial to the comprehensive evaluation and utilization of bioactive compounds in olive leaves.

Metagenomics reveals the formation mechanism of flavor metabolites during the spontaneous fermentation of potherb mustard (Brassica juncea var. multiceps).

Fermented vegetable flavors are closely associated with microbial metabolism. Here, shifts in flavor metabolites and their correlations to the structure and function of fermentative microbial communities were explored during the spontaneous fermentation process of potherb mustard (Brassica juncea var. multiceps), a traditionally fermented vegetable from China. Halophilic bacteria (HAB, i.e., Halomonas, Salinivibrio, and Vibrio) and lactic acid bacteria (LAB, i.e., Lactobacillus-related genera and Weissella) became highly abundant after potherb mustard fermentation. Further, HAB and LAB abundances exhibited significant, positive correlations with metabolites important in fermented potherb mustard flavoring (e.g., organic acids, amino acids, alcohols, aldehydes, and nitriles). Metagenomic analysis indicated that Halomonas, Salinivibrio, Weissella, and Lactobacillus-related genera were likely actively engaged in pyruvate metabolism (ko00620) and citrate cycle (TCA cycle, ko00020), leading to higher lactic and acetic acid concentrations, along with lower pH, which would affect levels of volatile isothiocyanates and nitriles that contribute to flavoring of fermented potherb mustard. Further, HAB and LAB were the primary populations inferred to be responsible for amino acid and fatty acid metabolism in addition to the biosynthesis of numerous volatile flavor compounds. This study highlights the predominance and importance of LAB and HAB during spontaneous fermentation of potherb mustard and provides new insights into their roles in this process.

Comparative Analysis of Traditional and Modern Fermentation for Xuecai and Correlations Between Volatile Flavor Compounds and Bacterial Community.

Differences in flavor compounds and bacterial communities of Xuecai by traditional and modern fermentation are poorly understood. Allyl isothiocyanate (E9), ethyl acetate (E1), 3-butenenitrile (N1), phenol (P1), ethanol (A1), and 3-(2,6,6-trimethyl-1-cyclohexen-1-yl) acrylaldehyde (L11) were the main flavor compounds that differed between Xuecai produced by traditional and modern fermentation. Among these compounds, the contents of N1 and E9 were higher in modern fermentation Xuecai. Traditional fermentation Xuecai possessed higher contents of A1, P1, E1, and L11. High-throughput sequencing showed that Lactobacillus-related genera was the most abundant genus (50%) in modern fermentation Xuecai. However, in traditional fermentation Xuecai, Halanaerobium (29.06%) and Halomonas (12.96%) were the dominant genera. Halophilic bacteria (HB) positively contribute to the flavor of Xuecai. Carbohydrate metabolism and amino acid metabolism were the most abundant pathways associated with the bacterial communities of the Xuecai. This indicated that Xuecai flavor formation is mainly dependent on protein and carbohydrate degradation. This study provides a novel insight that HB may be important for flavor formation of Xuecai.

Correlation of the bacterial communities with umami components, and chemical characteristics in Zhejiang xuecai and fermented brine.

Xuecai is a Chinese-fermented vegetable product that has attracted much attention because of its unique umami taste. However, investigations of the bacterial communities, umami components and chemical characteristics in xuecai and the brine have not yet been conducted. In this study, the bacterial communities and their correlations with the umami components and the chemical characteristics are investigated. The results showed that significantly higher contents of total protein, total acid, amino acid nitrogen, total sugar, titratable acidity, salt, amino acids and organic acids, as well as significantly lower nitrite concentrations were observed in the brine samples. The supervised pattern recognition method revealed that the xuecai and brine could be distinguished by these indicators. In addition, the xuecai and brine were both dominated by Lactobacillus, Halanaerobium and Halomonas. A correlation analysis showed that halophilic bacteria had a positive effect on the umami components and chemical characteristics of the xuecai and brine. However, lactic acid bacteria both showed a negative correlation with amino acids and most organic acids. Membrane transport, carbohydrate metabolism and amino acid metabolism were the most abundant pathways. These results enhance our understanding of the bacterial communities, as well as the umami components and chemical characteristics in Zhejiang xuecai and brine.

Daily Supplementation with Fresh Angelica keiskei Juice Alleviates High-Fat Diet-Induced Obesity in Mice by Modulating Gut Microbiota Composition.

SCOPE: Angelica keiskei is associated with several health benefits, but little is known about the effect of A. keiskei juice (AKJ), which is rich in polyphenols, coumarins, and other healthful agents, on high-fat diet-induced obesity or its relationship with intestinal microbiota composition changes. METHODS AND RESULTS: C57BL/6 mice are fed either a normal diet (ND group; n = 8), a high-fat diet (HFD group; n = 8), or a high-fat diet supplemented with AKJ (AKJ group; n = 8) for 10 weeks. The results show that AKJ prevents weight gain, lowered fat accumulation, blood glucose, serum lipid levels, hepatic steatosis, and modulates the level of expression of genes involved in lipid metabolism in mice with obesity. AKJ is found to normalize HFD-induced gut dysbiosis. Particularly, AKJ ameliorates HFD-dependent changes in the relative abundance of several taxa back to normal status (e.g., AKJ increased Bacteroides and decreased Mollicutes_RF9, Ruminococcaceae_UCG-014, Faecalibacterium, and Lactobacillus). Spearman's correlation analysis reveals that those genera are closely correlated with body weight, fasting serum glucose, and serum lipid levels. CONCLUSION: The results show that consumption of phytochemical-rich AKJ may prevent HFD-induced obesity and metabolic disorders via changes in metabolic genes and gut microbiota composition.

Extract of ice plant (Mesembryanthemum crystallinum) ameliorates hyperglycemia and modulates the gut microbiota composition in type 2 diabetic Goto-Kakizaki rats.

Ice plant (Mesembryanthemum crystallinum) extract (IPE) is a rich source of d-pinitol, which is widely known to have potential anti-diabetic effects. In the present study, response surface methodology (RSM) was used to optimize d-pinitol extraction conditions with the Box-Behnken design. We then evaluated the anti-diabetic effects properties of IPE that was extracted under optimized conditions (53 °C, 119 min extraction time, and 1 : 11 dilution) in type 2 diabetic Goto-Kakizaki (GK) rats. IPE (400 mg kg-1 day-1) effectively controlled the increased fasting blood glucose level (decreased by 45% vs. GK-control rats) and impaired glucose tolerance (decreased area under curve (AUC) of glucose values by 24%, p < 0.05 vs. GK-control rats) after eight weeks of treatment. Furthermore, IPE significantly improved pancreatic islet morphology, ß-cell survival, and insulin secretion in diabetic rats, thus contributing to the antihyperglycemic effect. Finally, prebiotic effects of IPE on gut microbiota were observed and included increased abundance of the beneficial bacteria Bacteroidales_S24-7 and Ruminococcaceae_UCG-014 and decreased abundance of Treponema_2 and Lactobacillus. Overall, IPE has a substantial effect on attenuating hyperglycemia and modulating gut microbiota composition in diabetic GK rats. Therefore, IPE might be a promising functional food for the prevention of diabetes.

Multifunctional supramolecular vesicles based on the complex of ferrocenecarboxylic acid capped pillar[5]arene and a galactose derivative for targeted drug delivery.

Supramolecular vesicles based on the host-guest complexation of ferrocenecarboxylic acid capped pillar[5]arene and a galactose derivative have been constructed, which showed dual-responsiveness and cancer cells targetability resulting from its ferrocenecarboxylic acid units and galactose units, respectively. This work provides a good example for the construction of multifunctional nanocarriers for targeted drug delivery.

Regioselective Benzoylation of Diols and Carbohydrates by Catalytic Amounts of Organobase.

A novel metal-free organobase-catalyzed regioselective benzoylation of diols and carbohydrates has been developed. Treatment of diol and carbohydrate substrates with 1.1 equiv. of 1-benzoylimidazole and 0.2 equiv. of 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) in MeCN under mild conditions resulted in highly regioselective benzoylation for the primary hydroxyl group. Importantly, compared to most commonly used protecting bulky groups for primary hydroxyl groups, the benzoyl protective group offers a new protection strategy.