Enzymatically Extracted Apple Pectin Possesses Antioxidant and Antitumor Activity

The biological activity of apple pectin extracted conventionally or enzymatically using endo-xylanase and endo-cellulase, was tested in vitro. The analyses were performerd in tetraplicates and the statistical significance of the differences were assessed using ANOVA, Tukey post hoc and LSD (the least significant difference) tests. Multivariate regression analysis was applied to determine the structural components that have a crucial importance for antioxidant and antitumor properties of pectins. The pectins extracted by enzymes contained up to four times more ferulic acid and showed twice as great ability to neutralize free radicals and Fe(III) reduction. The antiradical potential positively correlated with phenols, fucose and rhamnose content. In the assays performed on HT-29 human adenocarcinoma and B16F10 melanoma cell cultures, the “green” pectins, contrary to acid isolated ones, exhibited remarkable anti-neoplastic potential while being nontoxic to nontransformed L929 cell line. The pectins in the dose of 1 mg/mL were capable of inhibiting adhesion (max 23.1%), proliferation (max 40.4%), invasion (max 76.9%) and anchorage-independent growth (max 90%) of HT-29 cells (significance level p < 0.001). These pectin preparations were slightly less active towards B16F10 cells. The enzyme-isolated apple pectins may be useful as a functional food additive and an ingredient of the ointment formulas for post-surgical melanoma treatment.     

Pectin: A Bioactive Food Polysaccharide with Cancer Preventive Potential

Pectin is an acidic heteropolysaccharide found in the cell walls and the primary and middle lamella of land plants. To be authorized as a food additive, industrial pectins must meet strict guidelines set forth by the Food and Agricultural Organization and must contain at least 65% polygalacturonic acid to achieve the E440 level. Fruit pectin derived from oranges or apples is commonly used in the food industry to gel or thicken foods and to stabilize acid-based milk beverages. It is a naturally occurring component and can be ingested by dietary consumption of fruit and vegetables. Preventing long-term chronic diseases like diabetes and heart disease is an important role of dietary carbohydrates. Colon and breast cancer are among the diseases for which data suggest that modified pectin (MP), specifically modified citrus pectin (MCP), has beneficial effects on the development and spread of malignancies, in addition to its benefits as a soluble dietary fiber. Cellular and animal studies and human clinical trials have provided corroborating data. Although pectin has many diverse functional qualities, this review focuses on various modifications used to develop MP and its benefits for cancer prevention, bioavailability, clinical trials, and toxicity studies. This review concludes that pectin has anti-cancer characteristics that have been found to inhibit tumor development and proliferation in a wide variety of cancer cells. Nevertheless, further clinical and basic research is required to confirm the chemopreventive or therapeutic role of specific dietary carbohydrate molecules.     

改良的橘皮果胶抑制半乳糖凝集素-3与肿瘤相关功能的研究进展

半乳糖凝集素-3(Gal-3)是半乳糖凝集素家族中唯一一种嵌合体结构蛋白,含有一个特定的糖基识别结构域(CRD),能够特异性识别并结合半乳糖,此结构域可介导细胞与细胞之间的黏附和识别,并且能够促进部分肿瘤细胞的增殖和转移等。改良的橘皮果胶(MCP)可通过特异性结合肿瘤相关蛋白Gal-3糖基识别结构域,抑制Gal-3与其相关肿瘤受体的结合,阻断肿瘤细胞之间相关信号的传导,从而表现出较强的抗肿瘤活性。本文综述了MCP在Gal-3介导肿瘤发生发展过程中相关作用的研究进展。     

Deoxycholic Acid Modulates Cell-Junction Gene Expression and Increases Intestinal Barrier Dysfunction

Diet-related obesity is associated with increased intestinal hyperpermeability. High dietary fat intake causes an increase in colonic bile acids (BAs), particularly deoxycholic acid (DCA). We hypothesize that DCA modulates the gene expression of multiple cell junction pathways and increases intestinal permeability. With a human Caco-2 cell intestinal model, we used cell proliferation, PCR array, biochemical, and immunofluorescent assays to examine the impact of DCA on the integrity of the intestinal barrier and gene expression. The Caco-2 cells were grown in monolayers and challenged with DCA at physiological, sub-mM, concentrations. DCA increased transcellular and paracellular permeability (>20%). Similarly, DCA increased intracellular reactive oxidative species production (>100%) and accompanied a decrease (>40%) in extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathways. Moreover, the mRNA levels of 23 genes related to the epithelial barrier (tight junction, focal adhesion, gap junction, and adherens junction pathways) were decreased (>40%) in (0.25 mM) DCA-treated Caco-2 cells compared to untreated cells. Finally, we demonstrated that DCA decreased (>58%) the protein content of occludin present at the cellular tight junctions and the nucleus of epithelial cells. Collectively, DCA decreases the gene expression of multiple pathways related to cell junctions and increases permeability in a human intestinal barrier model.     

Dispersions of nanoclays of different shapes into aqueous and solid biopolymeric matrices. Extended physicochemical study

Dispersions of nanofillers into aqueous and solid biopolymeric matrices were studied from the physicochemical viewpoint. This work was carried out based on the idea that the combination of biopolymers, derived from renewable resources, and nanofiller, environmentally friendly, may form a new generation of nanomaterials with excellent and unique properties at low cost. To this purpose, two pectins with different degrees of methyl esterification and nanoclays like halloysite and laponite RD were selected. The thermodynamic and structural studies on the aqueous mixtures of pectin and nanoclay were able to discriminate the interactions, which control the adsorption of pectin onto the filler and the aggregation of both pectin and clay particles. The gained insights were useful to interpret the mesoscopic structure of the nanocomposites (prepared from the aqueous mixtures by means of the casting method) evidenced by SEM, thermal stability, tensile properties, and transparency investigations. The attained knowledge represents a basic point for designing new hybrid nanostructures in both the aqueous and the solid phase for specific purposes.     

Plant Polysaccharides Modulate Immune Function via the Gut Microbiome and May Have Potential in COVID-19 Therapy

Plant polysaccharides can increase the number and variety of beneficial bacteria in the gut and produce a variety of active substances, including short-chain fatty acids (SCFAs). Gut microbes and their specific metabolites have the effects of promoting anti-inflammatory activity, enhancing the intestinal barrier, and activating and regulating immune cells, which are beneficial for improving immunity. A strong immune system reduces inflammation caused by external viruses and other pathogens. Coronavirus disease 2019 (COVID-19) is still spreading globally, and patients with COVID-19 often have intestinal disease and weakened immune systems. This article mainly evaluates how polysaccharides in plants can improve the immune system barrier by improving the intestinal microecological balance, which may have potential in the prevention and treatment of COVID-19.     

Physicochemical,Thermal and Rheological Properties of Pectin Extracted from Sugar Beet Pulp Using Subcritical Water Extraction Process

The objective of this study was to characterize the properties of pectin extracted from sugar beet pulp using subcritical water (SWE) as compared to conventional extraction (CE). The research involved advanced modeling using response surface methodology and optimization of operational parameters. The optimal conditions for maximum yield of pectin for SWE and CE methods were determined by the central composite design. The optimum conditions of CE were the temperature of 90 °C, time of 240 min, pH of 1, and pectin recovery yield of 20.8%. The optimal SWE conditions were liquid-to-solid (L/S) ratio of 30% (v/w) at temperature of 130 °C for 20 min, which resulted in a comparable yield of 20.7%. The effect of obtained pectins on viscoamylograph pasting and DSC thermal parameters of corn starch was evaluated. The contents of galacturonic acid, degree of methylation, acetylation, and ferulic acid content were higher in the pectin extracted by SWE, while the molecular weight was lower. Similar chemical groups were characterized by FTIR in both SWE and CE pectins. Color attributes of both pectins were similar. Solutions of pectins at lower concentrations displayed nearly Newtonian behavior. The addition of both pectins to corn starch decreased pasting and DSC gelatinization parameters, but increased ΔH. The results offered a promising scalable approach to convert the beet waste to pectin as a value-added product using SWE with improved pectin properties.     

Exploring Effects of Chitosan Oligosaccharides on the DSS-Induced Intestinal Barrier Impairment In Vitro and In Vivo

Intestinal barrier dysfunction is an essential pathological change in inflammatory bowel disease (IBD). The mucus layer and the intestinal epithelial tight junction act together to maintain barrier integrity. Studies showed that chitosan oligosaccharide (COS) had a positive effect on gut health, effectively protecting the intestinal barrier in IBD. However, these studies usually focused on its impact on the intestinal epithelial tight junction. The influence of COS on the intestinal mucus layer is still poorly understood. In this study, we explored the effect of COS on intestinal mucus in vitro using human colonic mucus-secreted HT-29 cells. COS relieved DSS (dextran sulfate sodium)-induced mucus defects. Additionally, the structural characteristics of COS greatly influenced this activity. Finally, we evaluated the protective effect of COS on intestinal barrier function in mice with DSS-induced colitis. The results indicated that COS could manipulate intestinal mucus production, which likely contributed to its intestinal protective effects.     

Extraction of Pectin from Satsuma Mandarin Peel: A Comparison of High Hydrostatic Pressure and Conventional Extractions in Different Acids

Satsuma mandarin peel pectin was extracted by high hydrostatic pressure-assisted citric acid (HHPCP) or hydrochloric acid (HHPHP), and the physiochemical, structural, rheological and emulsifying characteristics were compared to those from conventional citric acid (CCP) and hydrochloric acid (CHP). Results showed that HHP and citric acid could both increase the pectin yield, and HHPCP had the highest yield (18.99%). Structural characterization, including NMR and FTIR, demonstrated that HHPHP showed higher Mw than the other pectins. The viscosity of the pectin treated with HHP was higher than that obtained with the conventional method, with HHPHP exhibiting significantly higher viscosity. Interestingly, all the pectin emulsions showed small particle mean diameters (D_4,3 being 0.2–1.3 μm) and extremely good emulsifying stability with centrifugation and 30-day storage assays, all being 100%. Satsuma mandarin peel could become a highly promising pectin source with good emulsifying properties, and HHP-assisted acid could be a more efficient method for pectin extraction.     

Structural patterns of rhamnogalacturonans modulating Hsp-27 expression in cultured human keratinocytes

Polysaccharide extracts were obtained from chestnut bran (Castanea sativa), grape marc (Vitis vinifera) and apple marc (Malus spp.) and fractionated by size exclusion chromatography after endopolygalacturonase degradation. Compositional and linkage analyses by GC and GC-MS showed the characteristic rhamnogalacturonan structure with specific arabinan (apple marc) and type II arabinogalactan (chestnut bran, grape marc) side chains. Type II arabinogalactan rhamnogalacturonan from chestnut bran significantly stimulated the in vitro differentiation of human keratinocytes, giving evidence of a tight structure-function relationship. This molecule comprises short and ramified 3- and 3,6-beta- D-galactan and 5- and 3,5-alpha-L-arabinan side chains, but also contains significant amounts of t-Xyl and 4-Xyl with a characteristic 2:1 ratio. Enzymatic hydrolysis of this polysaccharide produced fragments of lower molecular weight with unchanged xylose content which conserved the same ability to stimulate human keratinocyte differentiation. It could be then speculated that dimeric xylosyl-xylose and/or longer oligomeric xylose side chains attached to a galacturonan and closely associated to hairy rhamno-galacturonan domains are essential patterns that could determine the biological activity of pectins.     

Pectin-Based Formulations for Controlled Release of an Ellagic Acid Salt with High Solubility Profile in Physiological Media

Among bioactive phytochemicals, ellagic acid (EA) is one of the most controversial because its high antioxidant and cancer-preventing effects are strongly inhibited by low gastrointestinal absorption and rapid excretion. Strategies toward an increase of solubility in water and bioavailability, while preserving its structural integrity and warranting its controlled release at the physiological targets, are therefore largely pursued. In this work, EA lysine salt at 1:4 molar ratio (EALYS), exhibiting a more than 400 times increase of water solubility with respect to literature reports, was incorporated at 10% in low methoxylated (LM) and high methoxylated (HM) pectin films. The release of EA in PBS at pH 7.4 from both film preparations was comparable and reached 15% of the loaded compound over 2 h. Under simulated gastric conditions, release of EA from HM and LM pectin films was minimal at gastric pH, whereas higher concentrations—up to 300 μM, corresponding to ca. 50% of the overall content—were obtained in the case of the HM pectin film after 2 h incubation at the slightly alkaline pH of small intestine environment, with the enzyme and bile salt components enhancing the release. EALYS pectin films showed a good prebiotic activity as evaluated by determination of short chain fatty acids (SCFAs) levels following microbial fermentation, with a low but significant increase of the effects produced by the pectins themselves. Overall, these results highlight pectin films loaded with EALYS salt as a promising formulation to improve administration and controlled release of the compound.     

Pectin, a versatile polysaccharide present in plant cell walls

Pectin or pectic substances are collective names for a group of closely associated polysaccharides present in plant cell walls where they contribute to complex physiological processes like cell growth and cell differentiation and so determine the integrity and rigidity of plant tissue. They also play an important role in the defence mechanisms against plant pathogens and wounding. As constituents of plant cell walls and due to their anionic nature, pectic polysaccharides are considered to be involved in the regulation of ion transport, the porosity of the walls and in this way in the control of the permeability of the walls for enzymes. They also determine the water holding capacity. The amount and composition of pectic molecules in fruits and vegetables and other plant produce strongly determine quality parameters of fresh and processed food products. Pectin is also extracted from suitable agro-by-products like citrus peel and apple pomace and used in the food industry as natural ingredients for their gelling, thickening, and stabilizing properties. Some pectins gain more and more interest for their health modulating activities. Endogenous as well as exogenous enzymes play an important role in determining the pectic structures present in plant tissue, food products, or ingredients at a given time. In this paper functional and structural characteristics of pectin are described with special emphasis on the structural elements making up the pectin molecule, their interconnections and present models which envisage the accommodation of all structural elements in a macromolecule. Attention is also given to analytical methods to study the pectin structure including the use of enzymes as analytical tools.     

Influence of mechanical properties of pectin films on charge density and charge density distribution in pectin macromolecule

The hydration and mechanical properties of citrus pectin films were examined in conditions relevant to those in the plant cell wall. The pectins used for this study varied in the degree of esterification (DE) (high or low) and charge distribution on the backbone (random or block). The hydration of the films was controlled in an osmotic pressure experiment using polyethylene glycol solutions (PEG 20000). Hysteresis tests at constant deformation rate (stress vs deformation) were used for investigating the mechanical behaviour of films. Mechanical and hydration properties of pectin films were examined as a function of charge density, charge density distribution and counterion environment—K⁺, Ca²⁺, Mg²⁺. Swelling decreased with increasing counterion concentration. The effect is stronger in the case of Ca²⁺ and Mg²⁺ for low esterified pectins and therefore crosslinks from divalent ions could be assumed. The crosslink effect is confirmed in mechanical experiments where an increase in the film tensile modulus is observed with increasing counterion concentration. It is shown for the first time that in case of highly concentrated pectin solutions Mg²⁺ cations also act as a crosslinker for pectin macromolecules.     

Identifying ligand-binding specificity of the oligopeptide receptor OppA from Bifidobacterium longum KACC91563 by structure-based molecular modeling

As a ATP-binding cassette (ABC) transporter the OppA receptor plays key roles in protecting the host organism and transporting nutrients across the intestine by the oligopeptide transporter from symbiotic bacteria and directs maturation of the host immune system. Among lactic acid bacteria, Bifidobacterium longum KACC91563, isolated from fecal samples of healthy Korean neonates, has the capability to alleviate food allergy effects. Operating as a peptide importer, the extracellular OppA receptor from gram-positive B. longum KACC91563 translocates nutrients, specifically peptides, from the outside environment of the intestinal tract to the inside of symbiotic cells. In the present study we attempt to explicate the relationship between the substrate’s specificity from the OppA importer and the probiotic effects of B. longum KACC91563 in the host intestine. It was first identified in this study the specialized structure–function relationship from the OppA importer of B. longum KACC91563 with its structural and functional determinants. This could provide insights into substrate specificity of unique immunological properties and a key switch for the substrate’s metabolism to reprogramming immune responses in the host intestine by structure-based molecular modeling. The probiotic effects of oligopeptide substrate (such as a proline-rich peptide containing at least one branched residue of leucine, isoleucine, and valine) and its metabolism for the OppA from B. longum KACC91563 are attributed to enhancement of the epithelial barrier by several different strain specific pathways to prevent the strong adhesion of pathogens.     

Compositional heterogeneity in pectic polysaccharides: NMR studies and statistical analysis

Pectins are a class of heterogeneous polysaccharides used in the food industry as a result of their ability to form gels. They are primarily composed of a (1ŕ4)-α-D-galactopyranosyluronic acid backbone where the carboxylic acid group is methyl esterified at a level which depends on the source of the pectin and processing conditions used to isolate the material. Of considerable interest is the distribution of the free acid and methyl ester groups along this polymer chain. NMR spectroscopy, coupled with statistical analysis, is a powerful technique for the study of sequence distribution of monomers. Experimental conditions (temperature, pD) are reported which are appropriate for the analysis of pectic polysaccharides. Results are presented from a study of sequence distribution in native, modified, and fractionated pectins using ¹H and ¹³C solution NMR methods. The triad sequence information was examined using Bernoullian and Markovian probability models in conjunction with continuous Gaussian distribution and discrete models. Intermolecular heterogeneity in pectins results in triad frequencies which reflect the distribution of acid and ester monomers for native and base saponified pectins. Fractionation of pectin through physicochemical methods and subsequent spectroscopic analysis provide insight into this heterogeneity. Segregation into discrete polymer populations shows a structural diversity best approximated by a 3-component statistical model.     

Studies on pectin coating of liposomes for drug delivery

The present study investigated the surface coating of charged liposomes by three different types of pectin (LM, HM and amidated pectin) by particle size determinations and zeta potential measurements. The pectins and the pectin coated liposomes were visualized by atomic force microscopy. The adsorption of pectin onto positive liposomes yielded a reproducible increase in particle size and a shift of the zeta potential from positive to negative side for all three pectin types, whereas the adsorption of pectin onto negative liposomes did not render any significant changes probably due to electrostatic repulsion. The positive liposomes coated with HM-pectin gave the largest pectin coated particles with the least negative zeta potential, while the opposite was observed for the LM-pectin coated positive liposomes. Furthermore, results from dynamic light scattering revealed narrow size distributions, indicating that the degree of aggregation was low for the pectin coated liposomes. As liposomes are able to encapsulate drugs and pectin has been found to be mucoadhesive, these pectin coated liposomes may be potential drug delivery systems.     

Controlled Release of Sulfasalazine Release from “Smart” Pectin Gel Microspheres under Physiological Simulated Fluids

Sulfasalazine (SLZ) is a synthetic nonsteroidal anti-inflammatory drug used mainly for the treatment of an inflammatory bowel and other diseases. Two pectins with different methylation degrees were blended to synthesized gel microspheres by ionotropic gelation for SLZ encapsulation. The encapsulation efficiency was found to be around of 99% in all formulations tested. However, different SLZ release profiles related to the methylation degrees of pectin were observed. Mixture of low methylated (LM) and high methylated (HM) pectins in the presence of calcium(II) displayed the best microsphere morphologies among the formulations tested determined by optical and electronic microscopies. The percentage of drug release using a mixture of LM and HM pectins after 255?min in simulated gastric fluid (pH?=?1.2), simulated intestinal fluid (pH?=?6.8), and phosphate buffer (pH?=?7.4) were 15.0%, 47.0%, and 52.2%, respectively.     

Diffusing wave spectroscopy investigations of acid milk gels containing pectin

The influence of the polysaccharide pectin on the gelation of acidified milk is studied in concentrated, undiluted, quiescent systems, primarily using diffusing wave spectroscopy. For pectins with a low degree of methylesterification (DM), interactions with milk-serum calcium yielded precipitated polysaccharide aggregates, even without acidification, that subsequently did not interact with casein micelles. However, high DM fine structures do not interact significantly with serum-calcium and absorb onto casein micelles as the pH is reduced below 5. A limited surface coverage of high DM pectin facilitates efficient bridging which enhances the rate of micelle aggregation and subsequent gelation and produces a clear signature in the shape of the measured MSD. The work highlights the fact that the behaviour of pectin in milk systems depends not only on the interaction of different polymeric fine structures with casein micelles, but also to a large extent on the interactions with calcium.     

Effect of temperature on the intrinsic viscosity and conformation of different pectins

The effects of temperature on the intrinsic viscosity and on the conformation of pectin obtained from Citrus, Apple and Sunflower in a 0.17M NaCl solution were studied. Mark-Houwink plots for Orange, Apple and Sunflower pectin were obtained using HPSEC with online light scattering and viscosity detection. The intrinsic viscosity and flow activation energy E _a of pectin from the sources studied were measured over the temperature range 20–60°C. E _a values were 0.67, 0.69, 1.34, and 1.44 × 10⁷ J/(kmol) for commercial Citrus, Orange, Sunflower and Apple pectin, respectively. Intrinsic viscosity decreased linearly with increasing temperature, for all pectins except Apple one. These results clearly indicated that Apple pectin underwent structural changes that were more drastic than those that occurred for pectin from the other sources. E _a increased with decreasing weight average molar mass M _w indicating that pectin with low M _w were more asymmetric than pectin with higher values of M _w. Changes in the Huggins coefficients K _h with temperature for pectin from the various sources were attributed to the ability of pectin to aggregate, disaggregate and re-aggregate according to the temperature at which it was stored.     

Revision of adsorption models of xyloglucan on microcrystalline cellulose

Interactions among cellulose, hemicellulose and pectins are important for plant cell wall assembly and properties and also for industrial applications of these polysaccharides. Therefore, binding of pectin and xyloglucan on microcrystalline cellulose was investigated in this experiment by adsorption isotherms, zeta potential and scanning electron microscopy (SEM). Analysis of three isotherm models (Langmuir, Freundlich and Fowler-Guggenheim isotherms) showed that the experimental adsorption isotherm was well described via the Fowler-Guggenheim model, which includes lateral interaction between the adsorbate. The adsorption isotherm and zeta potential measurement showed that at temperature 25 °C only xyloglucan adsorbed on the microcrystalline cellulose. In case of xyloglucan on cellulose, the equilibrium was reached in about 3–4 h, and the kinetics of adsorption were well described by the multiexponential equation. Analysis of the model suggests that two steps can be distinguished: diffusion and reconformation in an adsorbed layer. No adsorption of pectin was observed in this study. SEM study showed that xyloglucan may prevent cellulose from aggregation.