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Journal of Agricultural and Food... Mar 2024Daidzein is a major isoflavone compound with an immense pharmaceutical value. This study applied a novel P450 CYP82D26 which can biosynthesize daidzein from...
Daidzein is a major isoflavone compound with an immense pharmaceutical value. This study applied a novel P450 CYP82D26 which can biosynthesize daidzein from (2)-naringenin. However, the recombinant P450 systems often suffer from low coupling efficiency, leading to an electron transfer efficiency decrease and harmful reactive oxygen species release, thereby compromising their stability and catalytic efficiency. To address these challenges, the SH3-GBD-PDZ (SGP) protein scaffold was applied to assemble a multienzyme system comprising CYP82D26, P450 reductase, and NADP-dependent aldehyde reductase in desired stoichiometric ratios. Results showed that the coupling efficiency of the P450 system was significantly increased, primarily attributed to the channeling effect of NADPH resulting from the proximity of tethered enzymes and the electrostatic interactions between NADPH and SGP. Assembling this SGP-scaffolded assembly system in yielded a titer of 240.5 mg/L daidzein with an 86% (2)-naringenin conversion rate, which showed a 9-fold increase over the free enzymes of the P450 system. These results underscore the potential application of the SGP-scaffolded multienzyme system in enhancing the coupling and catalytic efficiency of the P450 system.
Topics: NADP; NADPH-Ferrihemoprotein Reductase; Proteins; Isoflavones; Flavanones
PubMed: 38468401
DOI: 10.1021/acs.jafc.3c09854 -
Phytochemical Analysis : PCA Jul 2024Yangxinshi tablet (YXST) is a traditional Chinese medicine preparation characterized by its high efficacy and safety for the treatment of cardiovascular diseases....
INTRODUCTION
Yangxinshi tablet (YXST) is a traditional Chinese medicine preparation characterized by its high efficacy and safety for the treatment of cardiovascular diseases. Anionic compounds have been revealed as potential active components. However, there is currently limited research regarding its quality control.
OBJECTIVE
We aimed to establish a strategy for the simultaneous separation and determination of five key anionic compounds in YXST.
METHOD
A sensitive and efficient analytical method was developed and applied for the simultaneous separation and determination of five key compounds in YXST using large-volume sample stacking with polarity switching and micelle electrokinetic chromatography (LVSS-PS-MEKC) coupled with diode array detection. Crucial parameters, including sample volume, applied voltage, composition and pH of the running buffer, concentration of organic modifier, and switching time of the polarity, were systematically evaluated and optimized using a single variable method to enhance separation performance. Furthermore, the impact of cyclodextrin and sodium dodecyl sulfate as electrolyte modifiers was also investigated.
RESULTS
Under the optimal conditions, baseline separation of the five compounds (daidzein, puerarin, glycyrrhiztinic acid, chlorogenic acid, and salvianolic acid B) was achieved within 20 min. In comparison to the conventional MEKC mode, the constructed LVSS-PS-MEKC method exhibited a more than sixfold increase in the enrichment factor. The method was validated in terms of linearity, precision, accuracy, 24 h stability, and recovery and successfully applied to analyze YXST samples.
CONCLUSION
A sensitive strategy was developed for the simultaneous separation and determination of five key anionic components in YXST, offering a robust and efficient strategy for pharmaceutical analysis.
Topics: Chromatography, Micellar Electrokinetic Capillary; Tablets; Drugs, Chinese Herbal; Anions; Reproducibility of Results; Sodium Dodecyl Sulfate
PubMed: 38462488
DOI: 10.1002/pca.3347 -
The Journal of Physical Chemistry. B Mar 2024The effects that the main soy isoflavones, genistein and daidzein, have upon the biophysical properties of a model lipid bilayer composed of...
The effects that the main soy isoflavones, genistein and daidzein, have upon the biophysical properties of a model lipid bilayer composed of 1,2-dioleoyl--glycero-3-phosphocholine (DOPC) or DOPC with cholesterol (4 to 1 mol ratio) have been investigated by transbilayer water permeability, differential scanning calorimetry, and confocal Raman microspectroscopy. Genistein is found to increase water permeability, decrease phase transition temperature, reduce enthalpy of transition, and induce packing disorder in the DOPC membrane with an increasing concentration. On the contrary, daidzein decreases water permeability and shows negligible impact on thermodynamic parameters and packing disorder at comparable concentrations. For a cholesterol-containing DOPC bilayer, both genistein and daidzein exhibit an overall less pronounced effect on transbilayer water permeability. Their respective differential abilities to modify the physical and structural properties of biomembranes with varying lipid compositions signify a complex and sensitive nature to isoflavone interactions, which depends on the initial state of bilayer packing and the differences in the molecular structures of these soy isoflavones, and provide insights in understanding the interactions of these molecules with cellular membranes.
Topics: Genistein; Isoflavones; Lipid Bilayers; Cholesterol; Water
PubMed: 38417149
DOI: 10.1021/acs.jpcb.3c08390 -
Journal of Medicine and Life Nov 2023Ifosfamide (IFO), an alkylating chemotherapy agent, is known for its association with neurotoxicity and encephalopathy. This trial was designed to evaluate the...
Ifosfamide (IFO), an alkylating chemotherapy agent, is known for its association with neurotoxicity and encephalopathy. This trial was designed to evaluate the protective action of daidzein (DZN) against IFO-induced neurotoxicity in male rats by determining the difference in certain inflammatory and apoptotic markers in the brain tissue of rats. Twenty-eight Wistar rats, weighing 120-150 g, were divided into four groups of seven rats: Group 1 (Control) received no treatment; Group 2 was orally administered DZN (100 mg/kg/day) for seven days; Group 3 received a single intraperitoneal (IP) dose of IFO (500 mg/kg); Group 4 received oral DZN (100 mg/kg/day) for one week prior to a single IP dose of IFO on the seventh day. Twenty-four hours post-treatment, serum and brain tissue samples were collected for analysis. The results indicated a significant increase in serum inflammatory markers (TNF-alpha, IL-6, and iNOS) and the anti-inflammatory marker (IL-10), along with elevated caspase-3 enzyme activity in the brain tissue of the IFO-treated group compared to the control group. Conversely, pre-treatment with DZN significantly reduced serum inflammatory markers and caspase-3 levels in tissue. The findings suggest that daidzein has anti-inflammatory and anti-apoptotic properties, potentially offering protection against IFO-induced neurotoxicity in rats.
Topics: Rats; Male; Animals; Ifosfamide; Rats, Wistar; Neuroprotective Agents; Caspase 3; Antineoplastic Agents, Alkylating; Fanconi Syndrome; Anti-Inflammatory Agents; Isoflavones
PubMed: 38406792
DOI: 10.25122/jml-2023-0082 -
Journal of Food Science Apr 2024Soybean paste, a traditional fermented condiment, exhibits distinct quality attributes by its microbial communities. This study employed Illumina sequencing and LC-MS to...
Soybean paste, a traditional fermented condiment, exhibits distinct quality attributes by its microbial communities. This study employed Illumina sequencing and LC-MS to scrutinize the bacterial biota and metabolome of highly preserved (HP) and easily spoiled (ES) soybean pastes. Firmicutes were prevalent in both pastes, with HP showcasing greater microbial α-diversity compared to ES pastes. Bacillus predominated in HP pastes, whereas Lactobacillus was most abundant in ES pastes. Significant metabolic differences were observed between HP and ES samples in lipids, peptides, nucleic acids, secondary metabolite biosynthesis, protein digestion, amino acid metabolism, inflammatory mediator regulation, and neomycin, kanamycin, and gentamicin biosynthesis. Lactobacillus exhibited positive associations with daidzein and 3,4,5-trihydroxypentanoylcarnitine, whereas Bacillus showed negative correlations with 1,n6-ethenoadenosine, 2-deoxy-2,3-dehydro-n-acetyl-neuraminic acid, 3,4,5-trihydroxypentanoyl carnitine, and fructosyl valine. These findings highlight the collaborative impact of bacterial communities and metabolites on soybean paste quality attributes. This research enhances our comprehension of preservation mechanisms in fermented foods, particularly soybean pastes. PRACTICAL APPLICATION: The investigation would provide insights into the soybean pastes fermentation, safe and quality control methods, bio-preservative development strategies, and so on of soybean pastes for related studies and the consumers. Bacteria and their metabolites could be used to optimize the fermentation processes for the preservative and safe regulations.
Topics: Glycine max; Bacteria; Fermented Foods; Bacillus; Fermentation
PubMed: 38391005
DOI: 10.1111/1750-3841.16982 -
Analytical and Bioanalytical Chemistry Mar 2024Exposure to polyphenols is relevant throughout critical windows of infant development, including the breastfeeding phase. However, the quantitative assessment of...
Exposure to polyphenols is relevant throughout critical windows of infant development, including the breastfeeding phase. However, the quantitative assessment of polyphenols in human breast milk has received limited attention so far, though polyphenols may positively influence infant health. Therefore, a targeted LC-MS/MS assay was developed to investigate 86 analytes representing different polyphenol classes in human breast milk. The sample preparation consisted of liquid extraction, salting out, freeze-out, and a dilution step. Overall, nearly 70% of the chemically diverse polyphenols fulfilled all strict validation criteria for full quantitative assessment. The remaining analytes did not fulfill all criteria at every concentration level, but can still provide useful semi-quantitative insights into nutritional and biomedical research questions. The limits of detection for all analyzed polyphenols were in the range of 0.0041-87 ng*mL, with a median of 0.17 ng*mL. Moreover, the mean recovery was determined to be 82% and the mean signal suppression and enhancement effect was 117%. The developed assay was applied in a proof-of-principle study to investigate polyphenols in breast milk samples provided by twelve Nigerian mothers at three distinct time points post-delivery. In total, 50 polyphenol analytes were detected with almost half being phenolic acids. Phase II metabolites, including genistein-7-β-D-glucuronide, genistein-7-sulfate, and daidzein-7-β-D-glucuronide, were also detected in several samples. In conclusion, the developed method was demonstrated to be fit-for-purpose to simultaneously (semi-) quantify a wide variety of polyphenols in breast milk. It also demonstrated that various polyphenols including their biotransformation products were present in breast milk and therefore likely transferred to infants where they might impact microbiome development and infant health.
Topics: Female; Humans; Infant; Biological Monitoring; Chromatography, Liquid; Genistein; Glucuronides; Liquid Chromatography-Mass Spectrometry; Milk, Human; Polyphenols; Tandem Mass Spectrometry
PubMed: 38363307
DOI: 10.1007/s00216-024-05179-y -
Food & Function Mar 2024Daidzein, an isoflavone found abundantly in legumes, may benefit from bypassing upper gut absorption to reach the colon where it can be metabolized into the potent...
Daidzein, an isoflavone found abundantly in legumes, may benefit from bypassing upper gut absorption to reach the colon where it can be metabolized into the potent estrogen equol by the gut microbiome. To achieve this, we developed mucin coated protein-tannin multilayer microcarriers. Highly porous functionalized calcium carbonate (FCC) microparticles efficiently absorbed daidzein from a dimethyl sulfoxide solution, with a loading capacity of 21.6 ± 1.8 wt% as measured by ultra-high pressure liquid chromatography - mass spectrometry (UPLC-MS). Daidzein-containing FCC microparticles were then coated with a bovine serum albumin (BSA)-tannin -layer film terminated with mucin ((BSA-TA)-mucin) by layer-by-layer deposition from corresponding aqueous solutions followed by FCC decomposition with HCl. Raman spectroscopy confirmed mucin-tannin complexation involving both hydrophobic interactions and hydrogen bonding. The resulting multilayer microcarriers contained 54 wt% of nanocrystalline daidzein as confirmed by X-ray diffraction and UPLC-MS. Preliminary screening of several types of mucin coatings using an INFOGEST digestion model demonstrated that mucin type III from porcine stomach provided the highest protection against upper intestinal digestion. (BSA-TA)-mucin and (BSA-TA)-mucin microcarriers retained 71 ± 16.4% and 68 ± 4.6% of daidzein, respectively, at the end of the small intestinal phase. Mucin-free (BSA-TA) retained a lower daidzein amount of 46%. Daidzein release and further conversion into equol were observed during colonic studies with fecal microbiota from a healthy non-equol-producing donor and . The developed approach has potential for encapsulating other hydrophobic nutraceuticals or therapeutics, enhancing their bioaccessibility in the colon.
Topics: Equol; Chromatography, Liquid; Mucins; Tannins; Tandem Mass Spectrometry; Isoflavones; Polyphenols
PubMed: 38362621
DOI: 10.1039/d3fo03356b -
Endocrine Journal Apr 2024Soybean is a source of protein, fibers, and phytochemical isoflavones which are considered to have numerous health benefits for children and adulthood. On the other... (Review)
Review
Soybean is a source of protein, fibers, and phytochemical isoflavones which are considered to have numerous health benefits for children and adulthood. On the other hand, isoflavones are widely known as phytoestrogens that exert their action via the estrogen signaling pathway. With this regard, isoflavones are also considered as endocrine-disrupting chemicals. Endogenous estrogen plays a crucial role in brain development through binding to estrogen receptors (ERs) or G protein-coupled estrogen receptors 1 (GPER1) and regulates morphogenesis, migration, functional maturation, and intracellular metabolism of neurons and glial cells. Soy isoflavones can also bind to ERs, GPER1, and, furthermore, other receptors to modulate their action. Therefore, soy isoflavone consumption may affect brain development during the pre-and post-natal periods. This review summarizes the current knowledge on the mechanisms of isoflavone action, particularly in the early stages of brain development by introducing representative human, and animal models, and in vitro studies, and discusses their beneficial and adverse impact on neurobehavior. As a conclusion, the soy product consumption during the pre-and post-natal periods under proper range of dose showed beneficial effects in neurobehavior development, including improvement of anxiety, aggression, hyperactive behavior, and cognition, whereas their adverse effect by taking higher doses cannot be excluded. We also present novel research lines to further assess the effect of soy isoflavone administration during brain development.
Topics: Isoflavones; Humans; Brain; Signal Transduction; Animals; Glycine max; Phytoestrogens; Receptors, Estrogen; Receptors, G-Protein-Coupled
PubMed: 38346749
DOI: 10.1507/endocrj.EJ23-0314 -
Molecular Nutrition & Food Research Mar 2024The human gut microbiota regulates estrogen metabolism through the "estrobolome," the collection of bacterial genes that encode enzymes like β-glucuronidases and... (Review)
Review
The human gut microbiota regulates estrogen metabolism through the "estrobolome," the collection of bacterial genes that encode enzymes like β-glucuronidases and β-glucosidases. These enzymes deconjugate and reactivate estrogen, influencing circulating levels. The estrobolome mediates the enterohepatic circulation and bioavailability of estrogen. Alterations in gut microbiota composition and estrobolome function have been associated with estrogen-related diseases like breast cancer, enometrial cancer, and polycystic ovarian syndrome (PCOS). This is likely due to dysregulated estrogen signaling partly contributed by the microbial impacts on estrogen metabolism. Dietary phytoestrogens also undergo bacterial metabolism into active metabolites like equol, which binds estrogen receptors and exhibits higher estrogenic potency than its precursor daidzein. However, the ability to produce equol varies across populations, depending on the presence of specific gut microbes. Characterizing the estrobolome and equol-producing genes across populations can provide microbiome-based biomarkers. Further research is needed to investigate specific components of the estrobolome, phytoestrogen-microbiota interactions, and mechanisms linking dysbiosis to estrogen-related pathology. However, current evidence suggests that the gut microbiota is an integral regulator of estrogen status with clinical relevance to women's health and hormonal disorders.
Topics: Female; Humans; Phytoestrogens; Gastrointestinal Microbiome; Equol; Estrogens; Breast Neoplasms
PubMed: 38342595
DOI: 10.1002/mnfr.202300688 -
International Journal of Biological... Mar 2024This study aimed to prepare carrageenan/sodium alginate double-stabilized layers of zein nanoparticles loaded with daidzein using ultrasound technology to investigate...
This study aimed to prepare carrageenan/sodium alginate double-stabilized layers of zein nanoparticles loaded with daidzein using ultrasound technology to investigate the effect of ultrasound treatment on the stability of composite nanoparticles and encapsulation of daidzein. Compared with composite nanoparticles without ultrasound treatment, the encapsulation efficiency of nanoparticles was increased (90.36 %) after ultrasound treatment (320 W, 15 min). Ultrasound treatment reduced the particle size and PDI of nanoparticles and improved the stability and solubility of nanoparticles. Transmission electron microscopy (TEM) and scanning electron microscopy (SEM) revealed that the nanoparticles treated with ultrasound were smooth spherical and uniformly distributed. Fourier transform infrared spectroscopy (FTIR) results showed that the main forces that form nanoparticles are hydrogen bonding, electrostatic interactions and hydrophobic interactions. Fluorescence and CD chromatography showed that ultrasound treatment alters the secondary structure of zein and maintains nanoparticle stability. Encapsulation of daidzein in nanocarriers with ultrasound treatment can effectively scavenge DPPH and ABTS free radicals, improve antioxidant activity, and realize the slow release of daidzein in the gastrointestinal tract. The results showed that ultrasonication helps the construction of hydrophobic bioactives delivery carriers and provides better protection for unstable bioactives.
Topics: Zein; Carrageenan; Alginates; Nanoparticles; Particle Size; Isoflavones
PubMed: 38340944
DOI: 10.1016/j.ijbiomac.2024.130070