Binding Of Anthocyanins Research Articles

Exploring high hydrostatic pressure effects on anthocyanin binding to serum albumin and food-derived transferrins

This study investigated the effects of high hydrostatic pressure (HHP) on the binding interactions of cyanindin-3-O-glucoside (C3G) to bovine serum albumin, human serum albumin (HSA), bovine lactoferrin, and ovotransferrin. Fluorescence quenching revealed that HHP reduced C3G-binding affinity to HSA, while having a largely unaffected role for the other proteins. Notably, pretreating HSA at 500 MPa significantly increased its dissociation constant with C3G from 24.7 to 34.3 μM. Spectroscopic techniques suggested that HSA underwent relatively pronounced tertiary structural alterations after HHP treatments. The C3G-HSA binding mechanisms under pressure were further analyzed through molecular dynamics simulation. The localized structural changes in HSA under pressure might weaken its interaction with C3G, particularly polar interactions such as hydrogen bonds and electrostatic forces, consequently leading to a decreased binding affinity. Overall, the importance of pressure-induced structural alterations in proteins influencing their binding with anthocyanins was highlighted, contributing to optimizing HHP processing for anthocyanin-based products.

Antocianinas de la pitanga como fotosensibilizadores de celdas DSSC

Purified anthocyanin extracts were obtained from pitanga fruits (Eugenia Uniflora L.) and characterized by visible spectroscopy and FTIR, and the stability up to 85 ° C evaluated. These anthocyanins were mixed with different amounts of Mg2 +, Al3 +, Cr3+ at different working pH, finding the optimal complexation ratio for each system. The systems were also characterized by redox measurements, obtaining an oxidation potential close to 1 V for all cases, confirming the favourable characteristics of the compounds evaluated for their use in DSSC cells. FTIR confirmed the binding of anthocyanins and metal complexes to TiO2. The DSSC cells evaluated showed maximum conversion efficiency values of 0.24 % in complexes with chromium.

Black rice anthocyanins alleviate hyperuricemia in mice: Possible inhibitory effects on xanthine oxidase activity by cyanidin 3-O-glucoside

The aim of this study is to investigate the hypouricemic effect of black rice anthocyanins in vivo and the possible mechanism of their interaction with xanthine oxidase (XO). Four different secondary purified fractions were further prepared from the crude extract of black rice anthocyanins. In vitro enzyme inhibition experiments showed that fraction 2 indicated a reversible mixed inhibition of XO and the best inhibition. Fluorescence quenching spectroscopy and circular dichroism identified a single binding site within the active cavity of XO, the dominant molecular force was van der Waals, and the binding of anthocyanins to XO altered the standing secondary structure of the protein and inhibited the active center of the enzyme. Using a mouse model of hyperuricemia, fraction 2 and crude extract of black rice anthocyanins were found to have a strong ability to reduce uric acid levels and inhibit key enzymes in the uric acid production pathway, and to reduce kidney damage and oxidative stress. This study suggests that black rice anthocyanins have potential future applications as an effective anti-hyperuricemia bioactive substance.

Structure and composition of blueberry fiber pectin and xyloglucan that bind anthocyanins during fruit puree processing

Blueberry is a well-known source of antioxidants and prebiotic dietary fiber yet the fate of these health-beneficial ingredients during fruit processing it is not understood. Detailed composition and structure of blueberry pomace (PF) remaining after commercial blueberry fruit puree processing, as well as the water soluble and water insoluble fractions, were determined. The PF was comprised of dietary fiber (60.8%) that was largely insoluble (46.2% vs. 14.6% soluble dietary fiber) pectin, xyloglucan, arabinoxylan and mannan polysaccharides, fructose (11.22%) and glucose (10.37%) free sugars, protein (9%), fat (5%), and anthocyanins (6444.5 μg/g). The most abundant anthocyanins detected in PF were malvidin 3-arabinoside and peonidin 3-glucoside and the pectin was determined to be methyl-esterified and acetylated rhamnogalacturonan I with a 4,5-unsaturated function at the non-reducing end based on oligosaccharide structural analysis. Blueberry PF xyloglucan oligosaccharide side chain structures included xylose, arabinose and galactose consistent with arabino-xyloglucan structure. Microwave-assisted extraction of blueberry PF (pH 2 for 10 min at 80 °C) was performed to more clearly elucidate pectin structure, and revealed a pectin polymer with high molar mass (1072 kDa) and viscosity that was dependent upon the molar mass. A random coil shape was confirmed for both the blueberry PF and microwave extracted blueberry pectin, which agreed with previous reports that flexible blueberry pectin contributes to strong anthocyanin binding. In summary, the blueberry pomace anthocyanin content was slightly higher than that reported for fresh whole blueberry fruit, which suggests that most blueberry anthocyanins adhered to the pomace fiber, and most likely the insoluble pectin, during and after processing.

Anthocyanins decrease the internalization of TiO2 nanoparticles into 3D Caco-2 spheroids.

The influence of food components on nanoparticle (NP) internalization indicates a need to investigate the behaviors of NPs in a complex system. This study measured the changes of TiO2 NP colloidal stability and quenching of anthocyanin fluorescence to indicate NP-anthocyanin interactions, and cytotoxicity, oxidative stress, expression of ABC transporters and intracellular Ti concentrations in 3D Caco-2 spheroids co-exposed to NPs and anthocyanins to indicate the influence of anthocyanins on NP bio-effects. The anthocyanins were observed to have minimal impacts on colloidal properties of TiO2 NPs. Meanwhile, NP-anthocyanin co-exposure did not induce cytotoxicity or oxidative stress. The fluorescence quenching study indicated the binding of anthocyanins onto TiO2 NPs, and the binding affinity was inversely correlated with NP internalization into 3D Caco-2 spheroids. This may be partially related with the up-regulation of ABC transporters. Our results may provide novel insights into understanding the interactions of NPs and anthocyanins with human intestinal cells.

Fluorescence spectroscopy and molecular modeling of anthocyanins binding to bovine lactoferrin peptides

This work was aimed to obtain lactoferrin peptides, with anthocyanins-binding capabilities, by using eggplant peels extract as a source of anthocyanins. The chromatographic analysis of the extract evidenced the presence of five individual anthocyanins, with delfinidin-3-rutinoside being identified as the predominant. 20 small peptides were identified, from which four are containing Trp at C-terminal. By estimating the thermodynamic parameters, van der Waals and hydrogen bonding were found to have important roles in binding of anthocyanins to LF and LF-derived peptides. In order to complement the experimental results, the in silico methods were further employed to add single molecule level details on the potential interactions between different peptides and the main anthocyanins from eggplant peels. The docking tests indicated that the Trp containing peptides can bind, with different affinities either delphynidine-3-glycoside or delphynidine-3-rutinoside, therefore explaining the fluorescence quenching results. Our results have indicated a mechanism for the interactions between anthocyanins and LF and its small molecular weight peptides, whereas providing insights for formulating ingredients and foods with enhanced bioactives-binding properties.

The Binding mechanism of anthocyanins from sour cherries (Prunus cerasus L) skins to bovine β-lactoglobulin: A fluorescence and in silico-based approach

ABSTRACTFluorescence spectroscopy and computational methods were used to study the interaction between anthocyanins from sour cherries extract and bovine β-lactoglobulin. The experimental tests indicated that the number of binding sites was lower than 1, suggesting weak binding of anthocyanins by β-lactoglobulin, most probably as the result of the multitude of compounds present in the extract. Intrinsic and extrinsic fluorescence experiments have highlighted blue- and red-shifts in maximum emission. In agreement with the fluorescence spectroscopy findings, the in silico results suggested better affinity of the protein preliminary treated at high temperature for the anthocyanin. The molecular docking tests performed using the cyanidin 3-rutinoside as the ligand, one of the major components of anthocyanin extract indicated that the driving forces leading to complex formation mainly involve diamino and nonpolar neutral amino acids on the surface of the protein. The stability of the complex appeared to be mainly ensured by hydrophobic interactions and hydrogen bonds.

Functional and conformational changes to soy proteins accompanying anthocyanins: Focus on covalent and non-covalent interactions

The present study was aimed to evaluate the interaction between anthocyanins from black rice and soybean protein isolate (SPI) via non-covalent and covalent bindings and the impact of these interactions on the functional and conformational changes of soybean protein. The conformational changes of the protein structure with different concentrations of anthocyanins (0.05, 0.1%, and 0.2%) were analyzed using three-dimensional fluorescence and Fourier transform infrared spectroscopy. The anthocyanins were more likely to form covalent interactions with SPI instead of non-covalent interactions. The addition of anthocyanins changed the secondary structure of SPI with a decrease in β-sheets and an increase in β-turns and random coils. The emulsifying and foaming properties of SPI were improved after complexation with anthocyanins. This study might be useful for elucidating the mechanisms behind the binding of anthocyanins with SPI and the possible uses of SPI-anthocyanin complexes in food formulations.

Intermolecular binding of blueberry pectin-rich fractions and anthocyanin

Pectin was extracted from blueberry powder into three fractions of water soluble (WSF), chelator soluble (CSF) and sodium carbonate soluble (NSF). The fractions were incubated with cyanidin-3-glucoside (C3G), a mixture of five anthocyanidins (cyanidin, pelargonidin, malvidin, petunidin and delphinidin) or blueberry juice at pH 2.0–4.5. Free anthocyanins and bound anthocyanin–pectin mixtures were separated by ultrafiltration. WSF bound the least amount of anthocyanin at all pH values. CSF had stronger anthocyanin binding ability at pH 2.0–3.6, while NSF had stronger anthocyanin binding ability at pH 3.6–4.5. The pectin and anthocyanin binding was lowest at pH 4.5 and higher at pH 2.0–3.6. Nearly doubling C3G pigment content increased bound anthocyanin percentage by 16–23% at pH 3.6, which favored anthocyanin aromatic stacking, compared to 3–9% increase at pH 2.0. Ionic interaction between anthocyanin flavylium cations and free pectic carboxyl groups, and anthocyanin stacking may be two major mechanisms for pectin and anthocyanin binding.

Biopolymer nanoparticles as potential delivery systems for anthocyanins: Fabrication and properties

Biopolymer nanoparticles and microparticles may be used as delivery systems for bioactive compounds in food and pharmaceutical applications. In this study, biopolymer nanoparticles were assembled from whey protein isolate (WPI) and beet pectin (BP) using thermal processing and electrostatic complexation. This approach involved mixing the two biopolymers at pH5.8, heating (90°C, 5min) to induce protein nanoparticle formation and then adjusting the solution to pH4.0 to promote coating of the protein nanoparticles with pectin. This process led to the formation of relatively small anionic biopolymer nanoparticles (d<200nm) at pH4. The biopolymer particles had a high negative charged from pH8 to 4.0 but became less anionic at lower pH values. The mean particle diameter was relatively small (d<200nm) around pH4.0 but increased appreciably at lower and higher pH due to particle flocculation. The biopolymer nanoparticles were loaded with an anthocyanin-rich extract. A higher loading efficiency was observed when anthocyanin was added before heating the WPI-BP solution (LE=55%) rather than after (LE=35%), which was attributed to increased protein–polyphenol interactions. The encapsulation of anthocyanins within biopolymer particles improved their heat stability. However, encapsulated anthocyanin had a lower antioxidant activity than non-encapsulated anthocyanin, which was attributed to the thermal processing step required during particle fabrication and the binding of anthocyanins to biopolymers within the nanoparticles. Color measurements indicated that the encapsulation of anthocyanin within biopolymer particles did not inhibit its degradation after ascorbic acid addition. Overall, our results show that the encapsulation of anthocyanins within the biopolymer particles fabricated in this study was not particularly effective at improving their antioxidant activity or color stability. Alternative strategies are therefore needed to encapsulate this important color and nutraceutical agent.

Understanding the molecular mechanism of anthocyanin binding to pectin.

Association between anthocyanins and carbohydrates has drawn attention over the past few years and this interaction is of particularly importance in food chemistry since these compounds are often found together in plants and foodstuffs. This work intended to bring insights on the interaction between ionic carbohydrates (pectin) and two anthocyanins (cyanidin-3-O-glucoside, cy3glc and delphinidin-3-O-glucoside, dp3glc). The interaction between the flavylium cation and hemiketal anthocyanin forms was characterized by saturation transfer difference (STD) NMR spectroscopy and the respective dissociation constant (Kd) was obtained. This binding was also studied by Molecular Dynamics simulation. In the presence of the anthocyanin hemiketal form a weak interaction between anthocyanins and pectin seems to occur. A variation in the extent of this interaction was also noticed for the two anthocyanins with dp3glc bearing three hydroxyl groups, revealing to be a stronger binder to pectin (Kd ≈ 180 μM for dp3glc and Kd ≈ 250 μM for cy3glc). Experiments performed at acidic pH (flavylium cation) revealed a much stronger interaction (Kd ≈ 2 μM). These experimental results were also supported by theoretical studies which also revealed a stronger interaction in the presence of the anthocyanin flavylium cation and also a stronger interaction between pectin and dp3glc than with cy3glc (for the hemiketal form).

Interaction of anthocyanins with human serum albumin: Influence of pH and chemical structure on binding

The affinity of anthocyanins for human serum albumin (HSA) was determined by a fluorescence quenching method. The effects of pH and structure of anthocyanins on the binding constants were studied. The constants for binding of anthocyanins to HSA ranged from 1.08×105 to 13.2×105M−1. A hydrophobic effect at acidic pH was shown by the relatively high positive entropy values under the conditions studied. Electrostatic interactions, including hydrogen bonding, contributed to the binding at pH 7.4. The effect of structure of anthocyanins on the affinity was pH-dependent, particularly the effect of additional hydroxyl substituents. Hydroxyl substituents and glycosylation of anthocyanins decreased the affinity for binding to HSA at lower pH (especially pH 4), but increased the strength of binding at pH 7.4. In contrast, methylation of a hydroxyl group enhanced the binding at acidic pH, whilst this substitution reduced the strength of binding at pH 7.4. This paper shows that changes in anthocyanin structure or reductions in pH, which may occur in the region of inflammatory sites, have an effect on the binding of anthocyanins to HSA.

Lack of release of bound anthocyanins and phenolic acids from carrot plant cell walls and model composites during simulated gastric and small intestinal digestion

Separately, polyphenols and plant cell walls (PCW) are important contributors to the health benefits associated with fruits and vegetables. However, interactions with PCW which occur either during food preparation or mastication may affect bioaccessibility and hence bioavailability of polyphenols. Binding interactions between anthocyanins, phenolic acids (PAs) and PCW components, were evaluated using both a bacterial cellulose-pectin model system and a black carrot puree system. The majority of available polyphenols bound to PCW material with 60-70% of available anthocyanins and PAs respectively binding to black carrot puree PCW matter. Once bound, release of polyphenols using acidified methanol is low with only ∼20% of total anthocyanins to ∼30% of PAs being released. Less than 2% of bound polyphenol was released after in vitro gastric and small intestinal (S.I.) digestion for both the model system and the black carrot puree PCW matter. Confocal laser scanning microscopy shows localised binding of anthocyanins to PCW. Very similar patterns of binding for anthocyanins and PAs suggest that PAs form complexes with anthocyanins and polysaccharides. Time dependent changes in extractability with acidified methanol but not the total bound fraction suggests that initial non-specific deposition on cellulose surfaces is followed by rearrangement of the bound molecules. Minimal release of anthocyanins and PAs after simulated gastric and S.I. digestion indicates that polyphenols in fruits and vegetables which bind to the PCW will be transported to the colon where they would be expected to be released by the action of cell wall degrading bacteria.

Binding of polyphenols to plant cell wall analogues – Part 1: Anthocyanins

Anthocyanins are located within the vacuole of plant cells, and are released following cell rupture during eating or processing at which time they first come into contact with the plant cell wall. The extent of anthocyanin-cell wall interaction was investigated by monitoring the rate of anthocyanin depletion in the presence of pure cellulose or cellulose-pectin composites as cell wall models. It was found that anthocyanins interact with both cellulose and pectin over a two-stage process with initially (mins-hours) 13 similar to 18% of anthocyanins binding to cellulose or cellulose/pectincomposites. With prolonged exposure (days-weeks), a gradual increase in anthocyanin binding occurs, possibly due to anthocyanins stacking on top of a base layer. Binding of acylated and non-acylated anthocyanins followed a similar pattern with slightly more (5-10%) binding of the acylated forms. Composites with the highest pectin content had the greatest anthocyanin binding suggesting the existence of both ionic interactions (with pectin) and hydrophobic interactions (with cellulose) of anthocyanin with plant cell walls.

Interaction between gliadins and anthocyan derivatives

The interaction of gliadins with some anthocyanins (e.g. myrtillin, malvin, keracyanin, callistephin) and anthocyanidins (e.g. delphinidin, pelargonidin, cyanidin) has been analysed in aqueous solution at pH condition of the stomach, in which these compounds are initially metabolized.NMR, FT-IR and UV–Vis spectroscopic methods have been employed to determine the anthocyanin binding mode.The spectroscopic data seem to indicate that anthocyans are located along the polypeptide chains of gliadins in a generical molecular interaction between the two moieties. Our data do not exclude that hydrogen bonding interaction too is operating.Anthocyan–gliadins complexes are very soluble in acidic conditions.The results provide new insights into anthocyan–protein interaction and may have relevance to human health.