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#28910863   2017/09/14 Save this To Up

Acceleration of aglycone isoflavones and γ-aminobutyric acid production from doenjang using whole cell biocatalysis accompanied by protease treatment.

Recently, soy bean isoflavone aglycones (i.e. daidzein and genistein) and γ-aminobutyric acid (GABA) have begun to receive considerable consumer attention due to their potential as nutraceuticals. To produce these ingredients, multiple microorganisms and their enzymes are commonly used for catalysis in the nutraceutical industry. In this work, we introduce a novel fermentation process that uses whole cell biocatalysis to accelerate γ-aminobutyric acid and isoflavone aglycones production in doenjang (traditional Korean soybean paste). Microbial enzymes transform soybean isoflavone glycosides (i.e. daidzin and genistin) and monosodium glutamate into soy bean isoflavone aglycones and GABA. Lactobacillus brevis GABA 100 and Aspergillus oryzae KACC significantly reduced production time with the aid of a protease. The resulting levels of γ-aminobutyric acid and daidzein are higher, and genistein production resembled the levels in traditional doenjang fermented for over a year. Concentrations of γ-aminobutyric acid, daidzein, and genistein were measured as 7162 μg/g, 60 μg/g, and 59 μg/g, respectively on the seventh day of fermentation. Our results demonstrate that the administration of whole cell Lactobacillus brevis GABA 100 and Aspergillus oryzae KACC 40250 paired with a protease treatment is an effective method to accelerate γ-aminobutyric acid, daidzein, and genistein production in doenjang.

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#28887188   2017/09/09 Save this To Up

An extremely thermostable maltogenic amylase from Staphylothermus marinus: Bacillus expression of the gene and its application in genistin glycosylation.

The most extremely thermostable maltogenic amylase (SMMA) from archaeon Staphylothermus marinus has many potential applications in food processing. To ensure safety of microbial origin, a recombinant plasmid containing the enzymic gene and a constitutive promoter AmyR2 was constructed, and then transformed into a GRAS microorganism Bacillus subtilis. The purified SMMA from the liquid cultures of Bacillus has a specific activity of 66.96U/mg, two times more than that from Escherichia coli. SMMA was further employed to catalyze the genistion glycosylation using γ-CD as both glucosyl donors and solubilizer. Glycosylated genistins with one to four additional α-glucosyls and a molar percentage of 69.87% in genistin reaction mixture were identified and quantified by HPLC-UV-MS. The glycosylated genistins at 0.2-1.2mM showed an enhanced DPPH free radical scavenging capacity. To our knowledge, this is the first report on the Bacillus expression of archaeal maltogenic amylase.

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DNA (cytosine 5) methyltr Integrin β1 (CD29) Antib LPAM-1(Integrin α4, CD49 α-Internexin Antibody So INPP5F antibody Source Ra Interferon alpha-8 antibo Interferon alpha-6 antibo interleukin 17 receptor C Pancreatic Amylase antibo TCP-1 theta antibody Sour TGF beta induced factor 2 INPP1 antibody Source Rab

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#28873716   2017/09/06 Save this To Up

Characterization of four β-glucosidases acting on isoflavone-glycosides from Bifidobacterium pseudocatenulatum IPLA 36007.

Bifidobacterium pseudocatenulatum IPLA 36007 acts on isoflavone glycosides, releasing their corresponding aglycones. This strain-specific activity might be a key step in making isoflavones bioavailable and harnessing their oestrogenic activity. To investigate the molecular mechanisms involved in this activity, four glycosyl hydrolase-encoding genes in the IPLA 36007 genome (AW18_01575, AW18_09810, AW18_08145, and AW18_08090) were selected, synthesized with heterologous promoter and terminator signals (r-β-gluA, r-β-gluB, r-β-gluD and r-β-gluE, respectively), cloned into Escherichia coli, overexpressed as His-tagged proteins, and the enzymes purified and characterized. All four enzymes - GluAHis, GluBHis, GluDHis and GluEHis - proved to have β-glucosidase activity and deglycosylated (although at different rates) the isoflavone glycosides daidzin and genistin, releasing the aglycone moieties daidzein and genistein, respectively. GluDHis and GluEHis were also shown to hydrolyse β-glucosyl disaccharides such as cellobiose and gentiobiose, while GluAHis and GluBHis did not. Differences in activity were recorded for all four β-glucosidases at different pHs and temperatures under otherwise similar assay conditions, suggesting they have complementary activities under different environmental conditions. Two of the recombinant genes, r-β-gluA, and r-β-gluD, were cloned and expressed in the model lactic acid bacterium Lactococcus lactis, suggesting starter and probiotic organisms could be endowed with β-glucosidase activity. B. pseudocatenulatum IPLA 36007 contains additional β-glucosidases to those studied in this work, indicating a high level of redundancy for this enzymatic activity. Knowledge of glycoside-degrading enzymes should facilitate the development of novel, more effective or more selective prebiotics or functional foods for the promotion of bifidobacterial numbers in the human gut. It might also be of interest in the development of novel probiotics with specific health-promoting activities.

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#28852902   2017/08/30 Save this To Up

A new method to evaluate anti-allergic effect of food component by measuring leukotriene B4 from a mouse mast cell line.

Leukotrienes (LTs), chemical mediators produced by mast cells, play an important role in allergic symptoms such as food allergies and hay fever. We tried to construct an evaluation method for the anti-LTB4 activity of chemical substances using a mast cell line, PB-3c. PB-3c pre-cultured with or without arachidonic acid (AA) was stimulated by calcium ionophore (A23187) for 20 min, and LTB4 production by the cells was determined by HPLC with UV detection. LTB4 was not detected when PB-3c was pre-cultured without AA. On the other hand, LTB4 production by PB-3c pre-cultured with AA was detectable by HPLC, and the optimal conditions of PB-3c for LTB4 detection were to utilize the cells pre-cultured with 50 µM AA for 48 h. MK-886 (5-lipoxygenase inhibitor) completely inhibited LTB4 production, but AACOCF3 (phospholipase A2 inhibitor) slightly increased LTB4 production, suggesting that LTB4 was generated from exogenous free AA through 5-lipoxygenase pathway. We applied this technique to the evaluation of the anti-LTB4 activity of food components. PB-3c pre-cultured with 50 µM AA for 48 h was stimulated with A23187 in the presence of 50 µM soybean isoflavones (daidzin, genistin, daidzein, and genistein), equol, quercetin, or kaempferol. Genistein, equol, quercetin, and kaempferol strongly inhibited LTB4 production without cytotoxicity. These results suggest that a new assay system using PB-3c is convenient to evaluate LTB4 inhibition activity by food components. This method could be utilized for elucidation of the mechanisms of LTB4 release suppression by food components such as flavonoids and the structure-activity relationship.

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#28847347   2017/08/29 Save this To Up

Composition, Distribution, and Antioxidant Activity of Phenolic Compounds in 18 Soybean Cultivars.

Natural phenols are an important functional compound widely distributed in plants with benefits that promote human health. The content of total phenols, flavonoids, and anthocyanins and their composition distribution in 18 soybean cultivars was investigated. There are four phenolic acid distribution forms in these soybean cultivars, namely free, esterified, glycosided, and insoluble-bound. Total phenols, flavonoids, and anthocyanins from 6 black soybean cultivars were found in higher numbers than those from 12 other yellow soybean cultivars. Free and esterified phenolic acids were the main phenolic acid form in all 18 soybean samples. Chlorogenic acid and caffeic acid were the dominant phenolic acids in eight detected phenolic acids, and daidzin and genistin were the abundant isoflavones in five detected isoflavones. Furthermore, the antioxidant activities of total phenols from the 6 black soybean cultivars were greater than those from the 12 yellow soybean cultivars, and there was a significant positive correlation between antioxidant activity and total phenolic content. Black soybeans could be a potential resource for developing natural antioxidants that may play a crucial role in human health protection.

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#28791538   2017/08/09 Save this To Up

Attenuation of neurobehavioral and neurochemical abnormalities in animal model of cognitive deficits of Alzheimer's disease by fermented soybean nanonutraceutical.

The present study was performed to evaluate the efficacy of nanonutraceuticals (NN) for attenuation of neurobehavioral and neurochemical abnormalities in Alzheimer's disease. Solid-state fermentation of soybean with Bacillus subtilis was performed to produce different metabolites (nattokinase, daidzin, genistin and glycitin and menaquinone-7). Intoxication of rats with colchicine caused impairment in learning and memory which was demonstrated in neurobehavioral paradigms (Morris water maze and passive avoidance) linked with decreased activity of acetylcholinesterase (AChE). NN treatment led to a significant increase in TLT in the retention trials as compared to acquisition trial TLT suggesting an improved learning and memory in rats. Further, treatment of NN caused an increase in the activity of AChE (42%), accompanied with a reduced activity of glutathione (42%), superoxide dismutase (43%) and catalase (41%). It also decreased the level of lipid peroxidation (28%) and protein carbonyl contents (30%) in hippocampus as compared to those treated with colchicine alone, suggesting a possible neuroprotective efficacy of NN. Interestingly, in silico studies also demonstrated an effective amyloid-β and BACE-1 inhibition activity. These findings clearly indicated that NN reversed colchicine-induced behavioral and neurochemical alterations through potent antioxidant activity and could possibly impart beneficial effects in cognitive defects associated with Alzheimer's disease.

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#28771341   2017/08/03 Save this To Up

Soyasapogenol B and Genistein Attenuate Lipopolysaccharide-Induced Memory Impairment in Mice by the Modulation of NF-κB-Mediated BDNF Expression.

Lactobacillus plantarum C29-fermented defatted soybean (FDS), which contains soyasaponins such as soyasaponin I (SI) and soyasapogenol B (SB) and isoflavones such as genistin (GE) and genistein (GT), attenuated memory impairment in mice. Moreover, in the preliminary study, FDS and its soyasaponins and isoflavones significantly inhibited NF-κB activation in LPS-stimulated microglial BV2 cells. Therefore, we examined the effects of FDS and its constituents SI, SB, GT, and GE on LPS-induced memory impairment in mice. Oral administration of FDS (80 mg/kg), which has higher concentrations of SB and GE than DS, recovered LPS-impaired cognitive function in Y-maze (55.1 ± 3.5%) and passive avoidance tasks (50.9 ± 19.2 s) to 129.2% (74.1 ± 3.5%) and 114.2% (290.0 ± 22.4 s) of normal mice, respectively (P < 0.05). SB and GE (10 μM) also more potently attenuated LPS-impaired cognitive behavior than SI and GT, respectively. SB (10 mg/kg) was the most effective: treatment recovered LPS-impaired spontaneous alternation and latency time to 105.7% and 126.8% of normal control mice, respectively (P < 0.05). SB and GE significantly increased BDNF expression and CREB phosphorylation in LPS-treated mice and corticosterone-stimulated SH-SY5Y cells. Furthermore, SB and GE (10 μM) also significantly inhibited NF-κB activation in LPS-treated mice. These findings suggested that FDS and its constituent soyasaponins and isoflavones may attenuate memory impairment by the regulation of NF-κB-mediated BDNF expression.

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#28766479   2017/08/02 Save this To Up

An HPLC Method for the Determination of Isoflavones and the Evaluation of Their Antioxidant Capacity in Both Homogeneous and Microheterogeneous Systems.

In this work, we developed an HPLC method to simultaneously quantify and hence evaluate the stability, distribution, and antioxidant capacity of six isoflavones: genistein, genistin, daidzein, daidzin, glycitin, and biochanin A. Isoflavones have been described as having an important estrogenic activity to treat menopausal symptoms and can reduce postmenopausal bone loss and also participate in the prevention of cardiovascular diseases. These beneficial properties are believed derived from their capacity to act as free-radical scavengers. Isoflavones are formulated in capsules and creams and also can be used as antioxidants in liposomes. HPLC separation was achieved on an Agilent Hypersil ODS C18 column. The mobile phase consisted of 0.02-0.2% orthophosphoric acid in water-acetonitrile with gradient elution. The diode array detector was operated at 260 nm. The hydrophobicity of isoflavones was determined through their distribution in octanol-buffer. These results allowed us to establish a relation between chemical structure, pKa, lipophilicity, and the characteristics of the dispersion medium. Photolysis of hydrogen peroxide was used to measure the HO(•) scavenging capability of isoflavones. In liposomes, the order of reactivity of the studied compounds was genistein > biochanin A > genistin > daidzein > daidzin > glycitin.

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#28761823   2017/08/01 Save this To Up

Terminal Deoxynucleotidyl Transferase (TdT) Inhibiti on of Cord Blood Derived B and T Cells Expansion.

Purpose: Terminal deoxynucleotidyl transferase(TdT) is a DNA polymerase that is present in immature pre-B and pre-T cells. TdT inserts N-nucleotides to the V (D) J gene segment during rearrangements of genes, therefore, it plays a vital role in the development and variation of the immune system in vertebrates. Here we evaluated the relationship between cytokines like interleukin-2 (IL-2), interleukin-7 (IL-7), and interleukin-15 (IL-15) and TdT expression in cord blood mononuclear cells and also effect of inhibition in the expansion of B and T cells derived from cord blood. Methodes: The cord blood mononuclear cells were cultured with different combination of cytokines for 21days, which they were harvested in definite days (7, 14 and 21) and evaluated by flow cytometry. Results: Our data indicated that TdT expression increased in cord blood mononuclear cells using immune cell key cytokines without being dependent on the type of cytokines. TdT inhibition reduced both the expansion of B and T cells derived from cord blood and also declined the apoptosis and proliferation. Considered together, TdT played an important role in the control of the expansion of B and T cells derived from cord blood. Conclusion: considered together, it was observed that TdT expression was increased by cytokines and TdT inhibition not only reduced B and Tcells derived from cord blood, but it also affected the rate of apoptosis and proliferation.

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#28698467   2017/07/12 Save this To Up

Effect of Soy Isoflavones on Growth of Representative Bacterial Species from the Human Gut.

The present work aimed to assess the susceptibility of dominant and representative bacterial populations from the human gut to isoflavones and their metabolites. To do so, the minimum inhibitory concentration (MIC) of isoflavone glycosides, isoflavone aglycones, and equol to 37 bacterial strains was determined by broth microdilution. Additionally, for 10 representative strains, growth curves, growth rate (μ), and optical density (OD600 nm) of the cultures at 24 h were also determined. MICs of daidzin, genistin, daidzein, and genistein were >2048 μg mL(-1) for all strains assayed, while that of equol ranged from 16 μg mL(-1) for Bifidobacterium animalis subsp. animalis to >2048 μg mL(-1) for Enterobacteriaceae strains. Changes in growth curves, μ, and final OD were observed among the species in the presence of all tested compounds. Genistein reduced μ of Bacteroides fragilis, Lactococcus lactis subsp. lactis, and Slackia equolifaciens, while both genistein and equol increased that of Lactobacillus rhamnosus and Faecalibacterium prausnitzii. Compared to controls, lower final OD in the presence of aglycones and equol were recorded for some strains but were higher for others. Altogether, the results suggest that isoflavone-derived compounds could modify numbers of key bacterial species in the gut, which might be associated with their beneficial properties.

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