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Isoflavones (IFs) are biocompounds found in considerable amounts in soybean grains. However, to originate soybean-based materials, the grains must be subjected to numerous thermal and mechanical treatments, which can impair the IFs content. The influence of these treatments was investigated and a protocol for IFs extraction and quantification is proposed. Sequential extractions were performed on industrially pretreated soybean samples (broken, flakes, and collets), on okara submitted to different drying methods (freeze-drying, forced convection, and under vacuum), and on soybean oils extracted with hexane and ethanol. ß-glucosides levels were decreased by the thermal processes of lamination, expansion, and drying, while the aglycone forms were not affected. Lyophilization was identified as the most viable drying method for the quantification of IFs in okara. Soybean oils extracted with ethanol presented significant amounts of aglycone. Furthermore, two stages of extraction were sufficient for the recovery of the IFs from different matrices.
Assuntos
Isoflavonas , Glucosídeos , Glycine maxRESUMO
Soybean isoflavone aglycones have been investigated as potential wound healing compounds for topical application. The aim of this study was to evaluate the wound healing properties of a soybean isoflavone aglycones-rich fraction (IAF) when incorporated into lipid nanoemulsions dispersed in acrylic-acid hydrogels. Formulations exhibited a mean droplet size in the sub 200 nm range, negative ζ-potential (-60 mV), and displayed non-Newtonian pseudoplastic behavior. The addition of a gelling agent decreased the IAF release from formulations and improved the retention of these compounds in intact porcine ear skin when compared with a control propylene glycol solution. No IAF were detected in receptor fluid of Franz-type diffusion cells. However, increasing amounts of IAF were noticed in both skin layers and the receptor fluid when the tissue was partially injured (tape stripping), or when the epidermis was completely removed. In vitro studies showed that IAF elicits an increased proliferation and migration of keratinocytes (HaCaT cell line). Subsequently, the healing effect of the formulations was evaluated in a model of dorsal wounds in rats, by assessing the size of the lesions, histology, inflammatory markers, and antioxidant activity. Overall findings demonstrated the potential of IAF-loaded formulations to promote wound healing by increasing angiogenesis by â¼200 %, reducing the lipid oxidation (TBARS) by â¼52 % and the inflammation (TNFα) by â¼35 %, while increasing re-epithelialization by â¼500 %, visualized by the epithelium thickness.
Assuntos
Hidrogéis , Isoflavonas , Animais , Isoflavonas/farmacologia , Ratos , Pele , Glycine max , Suínos , CicatrizaçãoRESUMO
Soybean is one of the most important commodities in the world, being applied in feed crops and food, pharmaceutical industries in different ways. Soy is rich in isoflavones that in aglycone forms have exhibited significant anti-obesity and anti-lipogenic effects. Obesity is a global problem as several diseases have been related to this worldwide epidemic. The aim of this work was to verify the effect of free and immobilized ß-glucosidase, testing Lentikats, and sol-gel as carriers. Moreover, we wanted to examine if the different types of hydrolysis would generate extracts with distinct biological activity concerning lipid accumulation, PPAR-α regulation, and TNF-α, IL-6, and IL-10 concentrations using in vitro assays. Our results show that all formulations of ß-glucosidase could hydrolyze soy isoflavones. Thus, after 24 h of incubation, daidzein content increased 2.6-, 10.8-, and 12.2-fold; and genistein content increased 11.7, 11.4, and 11.4 times with the use of free enzyme, Lentikats®, and sol-gel immobilized enzyme, respectively. Moreover, both methodologies for enzyme immobilization led to promising forms of biocatalysts for application in the production of soy extracts rich in isoflavones aglycones, which are expected to bring about health benefits. A mild lipogenic effect was observed for some concentrations of extracts, as well as a slight inhibition in PPAR-α expression, although no significant differences were noticeable in the cytokines TNF-α, IL-10, and IL-6 as compared with the control.
Assuntos
Enzimas Imobilizadas/metabolismo , Glycine max/química , Iridoides/análise , Iridoides/farmacologia , Isoflavonas/química , Metabolismo dos Lipídeos/efeitos dos fármacos , beta-Glucosidase/metabolismo , Animais , Enzimas Imobilizadas/química , Hidrólise , Interleucina-10/metabolismo , Interleucina-6/metabolismo , Camundongos , Células RAW 264.7 , Fator de Necrose Tumoral alfa/metabolismo , beta-Glucosidase/químicaRESUMO
BACKGROUND: Urinary excretion of isoflavones and their metabolites after the consumption of soybean meal (SBM) and fermented soybean meal (FSBM) biscuits was investigated in humans in a randomized double-blinded crossover clinical trial. RESULTS: Aglycones were the most abundant metabolites in urine in both treatments. After FSBM consumption, total urinary excretion of aglycones and metabolites was 54% higher when compared to SBM biscuits. All metabolites were more rapidly excreted after FSBM biscuit consumption when compared to SBM biscuits. Urinary recovery of isoflavones was higher after FSBM biscuit consumption, reaching 67%. Women excreted higher amounts of colonic metabolites and lower amounts of aglycones than men. The prevalence of O-demethylangolensin-producer, equol-producer and nonproducer metabotypes was 56%, 11% and 36% after SBM consumption and 72%, 11% and 17% after FSBM consumption, respectively. CONCLUSIONS: Together, our findings indicate that fermentation improves isoflavone bioavailability and possibly reduces the impact of gut microbiota on the metabolism of isoflavones. © 2020 Society of Chemical Industry.
Assuntos
Glycine max/microbiologia , Isoflavonas/metabolismo , Adulto , Disponibilidade Biológica , Pão/análise , Feminino , Fermentação , Humanos , Masculino , Pessoa de Meia-Idade , Saccharomyces cerevisiae/metabolismo , Glycine max/metabolismo , Adulto JovemRESUMO
Glucosinolates are secondary plant metabolites of Brassicaceae. They exert their effect after enzymatic hydrolysis to yield aglycones, which become nitriles and epithionitriles through the action of epithiospecifier (ESP) and nitrile-specifier proteins (NSP). The mechanism of action of broccoli ESP and NSP is poorly understood mainly because ESP and NSP structures have not been completely characterized and because aglycones are unstable, thus hindering experimental measurements. The aim of this work was to investigate the interaction of broccoli ESP and NSP with the aglycones derived from broccoli glucosinolates using molecular simulations. The three-dimensional structure of broccoli ESP was built based on its amino-acid sequence, and the NSP structure was constructed based on a consensus amino-acid sequence. The models obtained using Iterative Threading ASSEmbly Refinement (I-TASSER) were refined with the OPLS-AA/L all atom force field of GROMACS 5.0.7 and were validated by Veryfy3D and ERRAT. The structures were selected based on molecular dynamics simulations. Interactions between the proteins and aglycones were simulated with Autodock Vina at different pH. It was concluded that pH determines the stability of the complexes and that the aglycone derived from glucoraphanin has the highest affinity to both ESP and NSP. This agrees with the fact that glucoraphanin is the most abundant glucosinolate in broccoli florets.
Assuntos
Brassica/química , Enzimas/química , Modelos Moleculares , Nitrilas/química , Proteínas de Plantas/química , Sequência de Aminoácidos , Isotiocianatos/química , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Estrutura Molecular , Conformação ProteicaRESUMO
ABSTRACTThe use of cell and plant tissue culture techniques to produce economically important active metabolites has been growing. Among these substances are total limonoid aglycones, which are produced by "pera" orange (Citrus sinensis (L.) Osbeck, Rutaceae) and have received considerable attention because of their anticancer actions. The main objective of the present study was to analyze and compare the levels of limonoid aglycones in seeds, callus cultures (originating from seeds), callus cultures (originating from hypocotyls), cell suspensions from hypocotyls cells, and cell suspensions from cotyledons. The cell cultures or C. sinensis were obtained by inoculating two strains of callus in MS medium supplemented with 2.0 µM 2,4-dichlorophenoxyacetic acid, 7.0 µM benzyl aminopurine, and 3% (w/v) sucrose in the dark. The highest concentrations of limonoid aglycone that were obtained were observed in cotyledon cell lines (240 mg/100 g dry weight) that were produced on day 21 of culture and hypocotyl cell lines on day 7 (210 mg/100 g dry weight). Explants of different origins under the same culture conditions had different limonoid aglycone content. The present results may suggest strategies for enhancing the productivity of biologically important limonoid aglycones and investigating the complex pathways of these secondary metabolites in plant tissue cultures.
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There is a growing interest in the pharmaceutical field concerning isoflavones topical delivery systems, especially with regard to their skin care properties and antiherpetic activity. In this context, the present work describes an ultra-fast liquid chromatography method (UFLC) for determining daidzein, glycitein, and genistein in different matrices during the development of topical systems containing isoflavone aglycones (IA) obtained from soybeans. The method showed to be specific, precise, accurate, and linear (0.1 to 5 µg mL(-1)) for IA determination in soybean acid extract, IA-rich fraction obtained after the purification process, IA loaded-nanoemulsions, and topical hydrogel, as well as for permeation/retention assays in porcine skin and porcine esophageal mucosa. The matrix effect was determined for all complex matrices, demonstrating low effect during the analysis. The stability indicating UFLC method was verified by submitting IA to acidic, alkaline, oxidative, and thermal stress conditions, and no interference of degradation products was detected during analysis. Mass spectrometry was performed to show the main compounds produced after acid hydrolysis of soybeans, as well as suggest the main degradation products formed after stress conditions. Besides the IA, hydroxymethylfurfural and ethoxymethylfurfural were produced and identified after acid hydrolysis of the soybean extract and well separated by the UFLC method. The method's robustness was confirmed using the Plackett-Burman experimental design. Therefore, the new method affords fast IA analysis during routine processes, extract purification, products development, and bioanalytical assays.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Genisteína/isolamento & purificação , Glycine max/química , Isoflavonas/isolamento & purificação , Administração Tópica , Animais , Transporte Biológico , Esôfago/efeitos dos fármacos , Esôfago/metabolismo , Furaldeído/análogos & derivados , Furaldeído/química , Furaldeído/isolamento & purificação , Furaldeído/farmacologia , Genisteína/química , Genisteína/farmacologia , Hidrogéis , Hidrólise , Isoflavonas/química , Isoflavonas/farmacologia , Mucosa/efeitos dos fármacos , Mucosa/metabolismo , Permeabilidade , Extratos Vegetais/química , Pele/efeitos dos fármacos , Pele/metabolismo , SuínosRESUMO
The aim of this study was to evaluate the profile of the contents of different forms of soybean isoflavones and the effect of germination time on these compounds and the physical parameters in BRS 284 soybean sprouts. Soybean seeds were germinated for 168 h, and the sprouts were collected every 24 h. The physical parameters and contents of different forms of isoflavones of the seeds and soybean sprouts were evaluated, and the data were subjected to regression analysis. The soybean seeds contained 26.0% ß-glucosides, 72.9% malonylglucosides and 1.2% aglycones. The yield of soybean sprouts was 632.4%. The effect of germination time was quadratic on the length, moisture and on the daidzin, genistin and genistein content; linear on the fresh weight and on the malonyldaidzin content. The dry matter and malonylglycitin content was constant, and glycitin and glycitein were not detected in the soybean sprouts.
Assuntos
Glycine max/química , Plântula/química , Sementes/química , Germinação , IsoflavonasRESUMO
The objective of this study was to investigate the immobilisation efficiency of soybean β-glucosidase (181.6 U/mL; 23.8 mg protein/mL) on activated chitosan beads. Central Composite Rotational Design (CCDR) 23 was used and the application of immobilised enzyme in commercial soy drink was evaluated. The activation of chitosan beads was achieved with established 2.5% glutaraldehyde, pH 7.5, 8 h incubation time (6 h with agitation and 2 h without agitation) at 37ºC. The highest immobilisation efficiency (%) of soybean β-glucosidase on chitosan beads obtained was 37.74 U/mL and 18.84 mg protein/4 chitosan beads at pH 7.5 and 20 h coupling time of enzyme-matrix (7 h with agitation and 13 h without agitation) at 4ºC. The immobilised enzyme incubated at 50ºC, pH 5.5 resulted in 24% increase in the aglycones content in commercial soy drink after 60 min.
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This study aimed to evaluate the antioxidant properties, bioactive compounds and other physico-chemical parameters from black soybean and its derivatives over 30 days under refrigeration at 4 °C and develop a probiotic petit suisse produced from black soybean. The soymilk showed the highest levels of isoflavones (109 mg/100 g), total phenolics (600 mg/100 g) and total anthocyanins (388 mg/100 g) with the highest response in the tests with DPPH⢠and ABTS+⢠on a dry basis. There was a significant increase (p ≤ 0.05) in antioxidant activity during storage due to the hydrolysis of isoflavone glycosides to aglycones in soymilk sample, having a strong linear correlation between the concentration of isoflavone aglycones and the antioxidant activity for ABTS+⢠(R = 0.9437, 0.9624 and 0.9992) and DPPH⢠(R = 0.9865, 0.9978 and 0.9911), respectively, for soymilk, quark and petit suisse. The conversion of isoflavone was influenced directly by the characteristics of each sample, inhibiting or promoting the action of the enzyme. The petit suisse developed is an alternative for consumers, providing isoflavones and anthocyanins, possessing probiotic average counts (108 CFU g⻹) during storage.