RESUMEN

Aim: Mitochondriogenesis refers to the process of creating and maintaining mitochondria, which plays an essential role in cellular metabolism. Mitochondrial processes such as energy generation, the response to oxidative stress, and cell death are all tightly regulated by enzymes. The flavonoid molecule malvidin-3-glucoside (M3G), which may be found in a wide variety of fruits and vegetables, has been shown to improve mitochondrial activity. However, the precise enzymes that mediate M3G's effect on mitochondriogenesis are yet unknown. Method: Here, we used in silico molecular modeling tools to look at how enzymes contribute to mitochondriogenesis after M3G administration. We used computational methods to discover candidate target enzymes known to interact with M3G and play important roles in mitochondrial physiology. Molecular docking was conducted to measure the binding affinity and stability of the M3G-enzyme complexes. The found enzymes' structural and functional features were analyzed using bioinformatics techniques, and the predicted functional implications of their interaction with M3G were formulated. Result: Our goal in doing these studies was to understand better how M3G regulates mitochondriogenesis by the action of altering SIRT-1, AMPK, and PGC-1α via M3G. Conclusion: In sum, our findings provide light on the molecular pathways by which M3G influences mitochondriogenesis. Furthermore, experimental validation of the discovered enzymes and their interactions with M3G may aid in the development of therapeutic approaches to improve mitochondrial function and cellular health.

RESUMEN

As one innate immune pattern recognition receptor, Toll-like receptor 4 (TLR4) recently has been considered as a critical player in glucolipid metabolism. Blueberries contain high level of anthocyanins, especially malvidin-3-glucoside (Mv-3-glc), which contribute the anti-inflammatory, hypoglycemic, and hypolipidemic effects. It is speculated that Mv-3-glc is able to possess these functions by binding to TLR4. Here, the noncovalent interactions of Mv-3-glc and TLR4 was explored through multi-techniques including fluorescence and ultraviolet-visible (UV-Vis) absorption spectroscopy, as well as molecular docking. The results demonstrated that Mv-3-glc was able to quench TLR4 intrinsic fluorescence effectively. A stable complex was formed spontaneously and the reaction was exothermic. The degree of binding of Mv-3-glc to TLR4 showed a strong dependence on the chemical concentration, temperature, and pH values. The negative signs for enthalpy (ΔH = -69.1 ± 10.8 kJ/mol) and entropy (ΔS = -105.0 ± 12.3 J/mol/K) from the interaction of the Mv-3-glc and TLR4 shows that the major driving forces are the hydrogen bonding and van der Waals' force, which is consistent with the molecular docking results. In addition, molecular docking predicted that the active center with specific amino acid residues, Phe126, Ser127, Leu54, Ile153, and Tyr131 was responsible for the site of Mv-3-glc binding to TLR4/myeloid differentiation protein-2 (MD-2). These findings confirmed that Mv-3-glc could bind to TLR4, which would be beneficial to understand the target therapeutic effects of blueberry anthocyanins on TLR4 in regulating glucolipid metabolism.

Asunto(s)

RESUMEN

Anthocyanins are important compounds in grapes and wine and significantly influence their characteristics. Ultraviolet light-C (UV-C) can be used as a tool to induce secondary metabolism, and in this study, it was used in 'Cabernet Sauvignon' grapes in the postharvest period to improve the anthocyanin profile and sensory attributes in wine produced with irradiated grapes. Grapes in the postharvest period were exposed to 0, 2, or 3 kJ m-2 UV-C radiation. After winemaking and storage time (6 months), physicochemical and color analyses, anthocyanin quantification and identification (HPLC-PDA-MS/MS), and sensory analyses were carried out. 'Cabernet Sauvignon' wine produced with grapes irradiated with 3 kJ m-2 UV-C showed increased spectrophotometric color, which is likely due to a 22.5% increase in total anthocyanin monomers, 59.3% of pyranoanthocyanins, 92.3% of direct condensation products, and 62.8% of acetaldehyde-mediated condensation products. In addition, this irradiated dose presented higher perception scores for visual color, aroma, taste, and was preferred by the tasters over the wine produced with non-irradiated grapes. This study is the first of its kind to show that the UV-C radiation treatment of grapes in the postharvest period can be a promising tool to improve the anthocyanin profile and sensorial quality of 'Cabernet Sauvignon' wine. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-021-05191-5.

RESUMEN

Anthocyanins are present in bright colored fruit and vegetables with growing evidence for their health benefits. Several methods exist in the literature to measure the total monomeric anthocyanin content in foods. Although the simplest method uses UV-Vis spectrophotometry, it requires the use of anthocyanin molar absorption coefficients (Ɛ). While commonly reported for some compounds, these values vary substantially between studies. This study collated and compared existing Ɛ values for a range of anthocyanin-3-glucosides, measured new Ɛ values for these compounds and underwent an inter-laboratory validation of spectrometry methods. The Ɛ values used for the determination of anthocyanin content in Australian blueberries, were shown to greatly affect the estimated total anthocyanin. Significant differences in the Ɛ values were observed when measured at 520 nm, or their absorbance maximum and substantial difference in the estimated total anthocyanins were observed when expressed as equivalent of cya-3-glu or mal-3-glu.

Asunto(s)

RESUMEN

The effect of extrusion cooking on bioactive compounds in third-generation snacks (TGSE) and microwave-expanded snacks (MWSE) prepared using black bean, blue maize, and chard (FBCS) was evaluated. FBCS was extruded at different moisture contents (MC; 22.2-35.7%), extrusion temperatures (ET; 102-142 °C), and screw speeds (SP; 96-171 rpm). Total anthocyanin content (TAC), contents of individual anthocyanins, viz., cyanidin-3-glucoside, malvidin-3-glucoside, pelargonidin-3-glucoside, pelargonidin-3-5-diglucoside, and delphinidin-3-glucoside chloride, total phenolic content (TPC), antioxidant activity (AA), and color parameters were determined. TAC and individual anthocyanin levels increased with the reduction in ET. ET and MC affected the chemical and color properties; increase in ET caused a significant reduction in TPC and AA. Microwave expansion reduced anthocyanin content and AA, and increased TPC. Extrusion under optimal conditions (29% MC, 111 rpm, and 120 °C) generated products with a high retention of functional compounds, with high TAC (41.81%) and TPC (28.23%). Experimental validation of optimized process parameters yielded an average error of 13.73% from the predicted contents of individual anthocyanins. Results suggest that the TGSE of FBCS obtained by combining extrusion and microwave expansion achieved significant retention of bioactive compounds having potential physiological benefits for humans.

RESUMEN

The grapevine red blotch disease (GRBD) was first noticed in 2008, impacting grape ripening. In general, GRBD reduces grape and wine quality resulting in significant economic losses. The purpose of the present study was to evaluate the effect of GRBD on agronomical parameters of 'Cabernet Sauvignon' vines at harvest. Using a metabolomics approach, the influence on primary and secondary metabolite profiling in skin + pulp/flesh and seeds were also determined. GRBD influenced °Brix and berry weight, as well as primary and secondary metabolites in both tissues. 1D 1H NMR was effective in quantifying the main primary and secondary metabolites affected by GRBD. RP-HPLC was similarly able to quantify the main phenolics affected. Multivariate analysis showed the influence of the virus on grape metabolites using both tools in two berry tissues. The effectiveness of both tools to describe sample variability was compared and the most affected metabolites in each tissue could be identified.

Asunto(s)

RESUMEN

Anthocyanin accumulation in grape berry skin is a crucial determinant of red/black grape berry quality. Recently, viticulture has been hampered by the issue of global warming, which has led to loss of anthocyanin accumulation in berry skin. The objectives of this study were to investigate the effect of allantoin on anthocyanin accumulation in berry skins of field-grown grapevines and to elucidate the molecular mechanism of the allantoin-induced anthocyanin accumulation in the berry skins. Allantoin enhanced anthocyanin accumulation in grape cultured cells and berry skins from field-grown grapevines at the early stage of ripening. MybA1 and UFGT, which encode the key transcription factor and enzyme in anthocyanin biosynthesis, were upregulated in allantoin-treated berry skins. Allantoin seems to increase the contents of delphinidin-based anthocyanin 3-glucosides in berry skins through the upregulation of F3'5'H gene that drives the synthesis of delphinidin-based anthocyanin 3-glucosides, compared with control berry skins. Allantoin increased soluble sugar contents in berries 10 days after treatment and upregulated abscisic acid (ABA)-responsible HT6 gene that encodes a key hexose transporter in sugar accumulation during ripening, in berry skins. These results suggested that physiological changes in allantoin-treated grape berries occurred in an ABA-dependent manner. Allantoin produced bioactive ABA through the ß-glucosidase-catalyzed hydrolysis of glucose-conjugated ABA but not through the ABA biosynthesis by NCED, a key enzyme in ABA biosynthesis, in berry skins. Allantoin application in viticulture under global warming conditions is expected to contribute to mitigating loss of red/black berry skin color.

Asunto(s)

RESUMEN

Protein-tyrosine phosphatase 1B (PTP1B, EC 3.1.3.48) is an important regulator of insulin signalling. Herein, we employed experimental and computational biology techniques to investigate the inhibitory properties of phenolics, identified from four in vitro gastrointestinal digested (IVGD) soft fruits, on PTP1B. Analysis by LC-MS/MS identified specific phenolics that inhibited PTP1B in vitro. Enzyme kinetics identified the mode of inhibition, while dynamics, stability and binding mechanisms of PTP1B-ligand complex were investigated through molecular modelling, docking, molecular dynamics (MD) simulations, and MM/PBSA binding free energy estimation. IVGD extracts and specific phenolics identified from the four soft fruits inhibited PTP1B (P < 0.0001) activity. Among the phenolics tested, the greatest inhibition was shown by malvidin-3-glucoside (P < 0.0001) and gallic acid (P < 0.0001). Malvidin-3-glucoside (Ki = 3.8 µg/mL) was a competitive inhibitor and gallic acid (Ki = 33.3 µg/mL) a non-competitive inhibitor of PTP1B. Malvidin-3-glucoside exhibited better binding energy than gallic acid and the synthetic inhibitor Dephostatin (-7.38 > -6.37 > -5.62 kcal/mol) respectively. Principal component analysis demonstrated malvidin-3-glucoside PTP1B-complex occupies more conformational space where critical WPD-loop displayed a higher degree of motion. MM/PBSA binding free energy for malvidin-3-glucoside to PTP1B was found to be higher than other complexes mediated by Van der Waals energy rather than electrostatic interaction for the other two inhibitors (-80.32 ± 1.25 > -40.64 ± 1.43 > -21.63 ± 1.73 kcal/mol) respectively. Altogether, we have established novel insights into the specific binding of dietary phenolics and have identified malvidin-3-glucoside as an PTP1B inhibitor, which may be further industrially developed for the treatment of type-2 diabetes.

Asunto(s)

RESUMEN

The detailed phenolic composition of different berry parts from two novel V. vinifera L. red grape genotypes (Moribel and Tinto Fragoso), together the well-known Tempranillo, was established using high performance liquid chromatography-diode array detection-electrospray ionization tandem mass spectrometry (HPLC-DAD-ESI-MS/MS) over two consecutive vintages (2016 and 2017). More than 50 phenolic compounds were identified and quantified: 25 anthocyanins, 17 flavonols, 7 hydroxycinnamic acid derivatives, 2 stilbenes, and several flavan-3-ols. As far as we know, some anthocyanin and flavonol dihexosides were reported for the first time in V. vinifera L. grapes. Application of Principal Component Analysis (PCA) to experimental data showed a good separation of the novel grape genotypes and Tempranillo according to the phenolic profile of skins and seeds, mainly based on the proportion of trisubstituted anthocyanin derivatives, flavonols and flavan-3-ols, being a useful tool to differentiate these grape varieties.

Asunto(s)

RESUMEN

The grape is an important fruit regarding economic and health benefit parameters, because of its large consumption around the world and their bioactive phenolic compounds. The drying process of BRS Morena grapes, whether pre-treated or not with olive oil for producing raisins, resulted in qualitative and quantitative changes in their phenolic composition (anthocyanins, flavonols, stilbenes, hydroxycinammic acid derivatives, flavan-3-ols and proanthocyanidins). The raisins with the pre-treatment preserved more anthocyanins and proanthocyanidins than the raisins not pre-treated. Moreover, the total dehydration time accelerated by approximately 40% in the raisins pre-treated. Therefore, the production of raisins of BRS Morena grapes pre-treated with olive oil as a natural surfactant constitutes an interesting process from both the industrial and health points of view, because of the remarkable reduction in the processing time and the preservation of high concentrations of flavonoids, which have important claims to health benefits from biological activities.

Asunto(s)

RESUMEN

This work describes the lipophilization reactions of malvidin 3-glucoside with different saturated fatty acid chain lengths using an enzymatic synthesis and the study of their physical-chemical and antioxidant properties. The lipophilic anthocyanins were obtained with satisfactory yields (22-40%) after column chromatography purifications, and they revealed the same appealing chromatic features of the parent anthocyanin. All of the compounds were characterized by mass spectrometry, confirming the regioselective acylation on the glucose moiety. The octanol-water partition coefficients and the hydrophobicity index of the different derivatives were determined, confirming a lipophilicity increase concomitant with the fatty acid chain length. The antioxidant profile was also evaluated by using two in vitro methods (ß-carotene-lineolate method and oxygen consumption assay). Overall, a maximum of antioxidant activity was achieved when malvidin 3-glucoside was conjugated with caprylic acid (C8). Altogether, the results obtained provide a good perspective for the technological application of these functionalized anthocyanins in the cosmetics and food industries.

Asunto(s)

RESUMEN

An efficient process for the purification of anthocyanin monomeric isomers from wild blueberries of Lake Saint-Jean region (Quebec, Canada) was developed and easy scalable at industrial purpose. The blueberries were soaked in acidified ethanol, filtered, and the filtrate was cleaned by solid phase extraction using silica gel C-18 and DSC-SCX cation-exchange resin. Anthocyanin-enriched elutes (87 wt.%) were successfully fractionated by preparative liquid chromatography. The major anthocyanins mono-galactoside, -glucoside and -arabinoside isomers of delphinidin, cyanidin, petunidin, peonidin and malvidin were isolated with a purity up to 100% according to their LC-MS and (1)H NMR spectra. The oxygen radical absorbance capacity (ORAC) of the obtained pure anthocyanins was evaluated. Delphinidin-3-galactoside has the highest capacity (13.062 ± 2.729 µmol TE/µmol), and malvidin-3-glucoside the lowest (0.851 ± 0.032 µmol TE/µmol). A mechanistic pathway preview is suggested for the anthocyanins scavenging free radical activity by hydrogen transfer.

Asunto(s)

RESUMEN

The main aim of this work was to develop an untargeted normal phase LC-MS method, starting from a targeted method already validated for the analysis of 135 polar metabolites. Since the LC instrument and column were the same, most of the chromatographic conditions remained identical, while the adaptations focused on maintaining the ionic strength of the eluents constant. The sample preparation was simplified and the effectiveness of LC-MS for long batches was evaluated, in order to record the maximum number of metabolites with good chromatographic resolution and the best MS stability and accuracy. The method was applied to study the influence of storage conditions on wine composition. Slightly sub-optimum storage conditions had a major impact on the polar metabolite fingerprint of the red wines analysed and the markers revealed included phenolics, vitamins and metabolites indentified in wine for the first time (4-amino-heptanedioic acid and its ethyl ester).

Asunto(s)

RESUMEN

The combined effect of anthocyanin-flavanol-flavonol ternary interactions on the colorimetric and chemical stability of malvidin-3-glucoside has been studied. Model solutions with fixed malvidin-3-glucoside/(+)-catechin ratio (MC) and variable quercetin-3-ß-d-glucoside concentration (MC+Q) and solutions with fixed malvidin-3-glucoside/quercetin-3-ß-d-glucoside ratio (MQ) and variable (+)-catechin concentration (MQ+C) were tested at levels closer to those existing in wines. Color variations during storage were evaluated by differential colorimetry. Changes in the anthocyanin concentration were monitored by HPLC-DAD. CIELAB color-difference formulas were demonstrated to be of practical interest to assess the stronger and more stable interaction of quercetin-3-ß-d-glucoside with MC binary mixture than (+)-catechin with MQ mixture. The results imply that MC+Q ternary solutions kept their intensity and bluish tonalities for a longer time in comparison to MQ+C solutions. The stability of malvidin-3-glucoside improves when the concentration of quercetin-3-ß-d-glucoside increases in MC+Q mixtures, whereas the addition of (+)-catechin in MQ+C mixtures resulted in an opposite effect.

Asunto(s)

RESUMEN

Nearly 20 million tons of winery by-products, with many biological activities, are discarded each year in the world. The extraction of bioactive compounds from Chenin Blanc, Petit Verdot, and Syrah grape by-products, produced in the semi-arid region in Brazil, was optimized by a Central Composite Rotatable Design. The phenolic compounds profile, antioxidant capacity against synthetic free radicals (DPPH and ABTS), reactive oxygen species (ROS; peroxyl radical, superoxide radical, hypochlorous acid), cytotoxicity assay (MTT) and quantification of TNF-α production in RAW 264.7 cells were conducted. Gallic acid, syringic acid, procyanidins B1 and B2, catechin, epicatechin, epicatechin gallate, quercetin 3-ß-d-glucoside, delfinidin 3-glucoside, peonidin 3-O-glucoside, and malvidin 3-glucoside were the main phenolic compounds identified. In general, rachis showed higher antioxidant capacity than pomace extract, especially for Chenin Blanc. All extracts showed low cytotoxicity against RAW 264.7 cells and Petit Verdot pomace suppressed TNF-α liberation in vitro. Therefore, these winery by-products can be considered good sources of bioactive compounds, with great potential for application in the food and pharmaceutical industries.

Asunto(s)

RESUMEN

Three Primitivo (Vitis vinifera, cv.) red wines were microvinified by means of different winemaking technologies: no stem-contact fermentation destemming 100% of grapes (D100); stem-contact fermentation destemming 75%, 50% of grapes (D75-D50) respectively. The objectives of this work were to improve proanthocyanidins content in wine, to monitor the relationships between anthocyanins/tannins and to detect the effects on the polymerization state of polyphenols after 12 months storage of wines. D100 showed higher anthocyanins content but lower color intensity and phenolic compounds content with respect to the theses D75 and D50, the last two showing lower anthocyanins content due their partial adsorption by grape stems during the fermentation. D75 gave the best results in terms of anthocyanins/color intensity balance and showed a better wine tannin component with respect to D50. Moreover after 12 months storage D50 reached a more advanced and stable polymerization state of colored pigments than the other wines.

Asunto(s)

RESUMEN

Field studies were conducted in Puglia (Italy) to evaluate the influence of defoliation around cluster zones on grape and wine quality. Nero di Troia grapes were subjected to four different treatments: N: no leaf removal; E: leaf removal in the area of the clusters along the east side (at complete veraison); E/W: leaf removal in the area of the clusters along the east and west side (at complete veraison); and F: almost complete leaf removal along the west side (at complete veraison) and at pre-harvest also along the east side. Grapes of defoliated vines generally showed higher sugar content, lower titratable acidity, total flavonoids, flavonoids different from anthocyanins, and total phenolic content than grapes from non-defoliated vines while their total anthocyanin concentration was not affected by defoliation at a significant level. Concerning wines, alcohol content, residual soluble solids, different forms of anthocyanins but also volatile acidity were generally higher in samples from defoliated vines. Differences were also highlighted among the defoliation treatments: the best results in terms of dry matter, sugar and alcohol content were observed in the samples submitted to the more severe defoliation as a consequence of the higher light availability and berry temperature. Concerning the concentration of the individual phenolics, significant differences were highlighted for: caffeic and caftaric acids, peonidin- and malvidin-3-p-coumaroylglucoside, which were higher in the E wines; quercetin-3-glucoside, galactoside, and rhamnoside, and procyanidins, which were higher in F wines.

RESUMEN

Although phenolics are recognized to be related with health benefits by limiting lipid oxidation, in wine, they are the primary substrates for oxidation resulting in the quinone by-products with the participation of transition metal ions. Nevertheless, high quality Port wines require a period of aging in either bottle or barrels. During this time, a modification of sensory properties of wines such as the decrease of astringency or the stabilization of color is recognized to phenolic compounds, mainly attributed to anthocyanins and derived pigments. The present work aims to illustrate the oxidation of red Port wine based on its phenolic composition by the effect of both thermal and oxygen exposures. A kinetic approach toanthocyanins degradation was also achieved. For this purpose a forced red Port wine aging protocol was performed at four different storage temperatures, respectively, 20, 30, 35 and 40°C, and two adjusted oxygen saturation levels, no oxygen addition (treatment I), and oxygen addition (treatment II). Three hydroxycinnamic esters, three hydroxycinnamic acids, three hydroxybenzoic acids, two flavan-3-ols, and six anthocyanins were quantitated weekly during 63days, along with oxygen consumption. The most relevant phenolic oxidation markers were anthocyanins and catechin-type flavonoids, which had the highest decreases during the thermal and oxidative red Port wine process. Both temperature and oxygen treatments affected the rate of phenolic degradation. In addition, temperature seems to influence mostly the phenolics kinetic degradation.

RESUMEN

The acylation of procyanidin B4 with a saturated fatty acid chloride containing 18 carbon atoms was studied in order to obtain procyanidin B4 3-O-di-stearic acid conjugate. This compound was structurally characterised by mass spectrometry and 1D and 2D NMR techniques. Derivatization of malvidin-3-glucoside using stearoyl chloride in acetonitrile was also performed yielding mono-, di- and tri-stearic ester derivatives. The novel derivatives obtained revealed significant antioxidant activity, although lower than the respective precursors. However, the chemical modification of anthocyanins and procyanidins (water soluble pigments) to more lipophilic compounds has the advantage of increased bioavailability in biological matrices, and to potentiate their application in food matrices and cosmetic products.

Asunto(s)

RESUMEN

Anthocyanin degradation has been proposed as one of the primary causes for reduced colour and quality in red wine grapes grown in a warm climate. To study anthocyanin degradation we infused berries with L-phenyl-(13)C6-alanine and then tracked the fate of the anthocyanins comparing normal (25 °C) and warm (45 °C) temperature conditions. An untargeted metabolomics approach was aided by filtering the MS data using software algorithms to extract all M and M+6 isotopic peak pairs, allowing the analysis to focus solely on the metabolites of phenylalanine. A paired-comparison t-test was performed over the 8 biological replicates revealing 13 metabolites that were statistically different between 25 °C and 45 °C treatments. Most of these features had lower abundances in 45 °C samples, confirming that 45 °C treatment caused anthocyanin degradation. In addition, resveratrol was significantly reduced following heat treatment. However, 5 metabolites increased following the 45 °C treatment. These unidentified metabolites are therefore suspects for anthocyanin degradation products.

Asunto(s)