The Antidiabetic Effect of Grape Pomace Polysaccharide-Polyphenol Complexes

Type 2 diabetes mellitus (T2DM) is one of the most prevalent chronic metabolic diseases of the 21st century. Nevertheless, its prevalence might be attenuated by taking advantage of bioactive compounds commonly found in fruits and vegetables. This work is focused on the recovery of polyphenols and polysaccharide–polyphenol conjugates from grape pomace for T2DM management and prevention. Bioactives were extracted by solid–liquid extraction and by pressurized hot water extraction (PHWE). Polyphenolic fraction recovered by PHWE showed the highest value for total phenolic content (427 mg GAE.mg−1), mainly anthocyanins and proanthocyanidins, and higher antioxidant activity compared to the fraction recovered by solid–liquid extraction. Polysaccharide–polyphenol conjugates comprehended pectic polysaccharides to which approximately 108 mg GAE of phenolic compounds (per mg fraction) were estimated to be bound. Polyphenols and polysaccharide–polyphenol conjugates exhibited distinct antidiabetic effects, depending on the extraction methodologies employed. Extracts were particularly relevant in the inhibition of a-glucosidase activity, with free polyphenols showing an IC50 of 0.47 mg.mL−1 while conjugates showed an IC50 of 2.7, 4.0 and 5.2 mg.mL−1 (solid–liquid extraction, PHWE at 95 and 120°C, respectively). Antiglycation effect was more pronounced for free polyphenols recovered by PHWE, while the attenuation of glucose uptake by Caco-2 monolayers was more efficient for conjugates obtained by PHWE. The antidiabetic effect of grape pomace bioactives opens new opportunities for the exploitation of these agri-food wastes in food nutrition, the next step towards reaching a circular economy in grape products.

ISOLATION AND PHYSICOCHEMICAL CHARACTERIZATION OF PECTIN FROM NONCONVENTIONAL NATU-RAL RAW MATERIALS

Obtaining and studying the functional properties and determining the subsequent purpose in the practical use of pectin substances requires a thorough study of physical and chemical characteristics of the polysaccharide. By using the acidic hydrolysis method, pectic polysaccharides were isolated from nonconventional raw materials of natural origins - basidial fungus mushrooms Fomes fomentarius, Ganoderma lucidum, Inonotus hispidus, Polýporus squamosus. The polysaccharides isolated were characterized: such as content of free and etherifying carboxyl groups, total -СООН groups, -ОСН3 groups, degree of etherification and by data of the IR-spectroscopy. The pectic polysaccharides isolated in identical conditions from the basidial mushrooms were powders with colors from light yellow to is light-brown by the appearance, possess sub-acidic taste, have no extraneous smack and a smell. The pectin from Inonotus hispidus was much lighter than other pectins (Fomes fomentarius, Ganoderma lucidum, Polýporus squamosus). The pectin’s isolated were similar to the apple pectin by the organoleptic properties. They can classified as averagely etherified pectins regarding the degree etherification. Etherification degree of the studied pectins was in the range of 50.00–66.67%. The methoxyl groups were ranged between 8.47–11.43%. Content of free carboxylic groups in the range of 2.25–5.40%.

Isolation, Chemical Characterization and Antioxidant Activity of Pectic Polysaccharides of Fireweed (Epilobium angustifolium L.)

The aim of this study was to isolate pectins with antioxidant activity from the leaves of Epilobium angustifolium L. Two pectins, EA-4.0 and EA-0.8, with galacturonic acid contents of 88 and 91% were isolated from the leaves of E. angustifolium L. by the treatment of plant raw materials with aqueous hydrochloric acid at pH 4.0 and 0.8, respectively. EA-4.0 and EA-0.8 were found to scavenge the DPPH radical in a concentration-dependent manner at 17–133 μg/mL, whereas commercial apple pectin scavenged at 0.5–2 mg/mL. The antioxidant activity of EA-4.0 was the highest and exceeded the activity of EA-0.8 and a commercial apple pectin by 2 and 39 times (IC50—0.050, 0.109 and 1.961 mg/mL), respectively. Pectins EA-4.0 and EA-0.8 were found to possess superoxide radical scavenging activity, with IC50s equal to 0.27 and 0.97 mg/mL, respectively. Correlation analysis of the composition and activity of 32 polysaccharide fractions obtained by enzyme hydrolysis and anionic exchange chromatography revealed that the antioxidant capacity of fireweed pectins is mainly due to phenolics and is partially associated with xylogalacturonan chains. The data obtained demonstrate that pectic polysaccharides appeared to be bioactive components of fireweed leaves with high antioxidant activity, which depend on pH at their extraction.

The Role of Pectobacterium atrosepticum Exopolysaccharides in Plant–Pathogen Interactions

The phytopathogenic bacterium Pectobacterium atrosepticum (Pba), one of the members of the soft rot Pectobacteriaceae, forms biofilm-like structures known as bacterial emboli when colonizing the primary xylem vessels of the host plants. The initial extracellular matrix of the bacterial emboli is composed of the host plant’s pectic polysaccharides, which are gradually substituted by the Pba-produced exopolysaccharides (Pba EPS) as the bacterial emboli “mature”. No information about the properties of Pba EPS and their possible roles in Pba-plant interactions has so far been obtained. We have shown that Pba EPS possess physical properties that can promote the maintenance of the structural integrity of bacterial emboli. These polymers increase the viscosity of liquids and form large supramolecular aggregates. The formation of Pba EPS aggregates is provided (at least partly) by the acetyl groups of the Pba EPS molecules. Besides, Pba EPS scavenge reactive oxygen species (ROS), the accumulation of which is known to be associated with the formation of bacterial emboli. In addition, Pba EPS act as suppressors of the quantitative immunity of plants, repressing PAMP-induced reactions; this property is partly lost in the deacetylated form of Pba EPS. Overall, our study shows that Pba EPS play structural, protective, and immunosuppressive roles during Pba–plant interactions and thus should be considered as virulence factors of these bacteria.

Effect of the Olive Oil Extraction Process on the Formation of Complex Pectin–Polyphenols and Their Antioxidant and Antiproliferative Activities

The aim of this research was to investigate the interaction of phenols and pectic polysaccharides during the olive oil extraction process. For this, pectin was extracted from fresh olive fruits and compared to the pectin isolated from the paste resulting from the extraction of the olive oil after milling with malaxation at 30 °C/30 min and subsequent centrifugation of the olive paste from the same lot of olive fruits in a system called ABENCOR (AB). The results indicate that these interactions were enhanced during the olive oil extraction process. In addition, the resulting AB extracts exhibited high antioxidant activity (ORAC) and strong antiproliferative activity in vitro against colon carcinoma Caco-2 cell lines compared to olive fruit extracts. The polyphenols associated mainly with the acidic pectin substance, with a higher content in AB extracts, seem to be responsible for these activities, and appear to maintain their activities in part after complexation. However, even in olive fruit extracts with smaller amounts of phenols in their compositions, pectic polysaccharides may also be involved in antioxidant and antiproliferative activities.

Mapping Pectic-Polysaccharide Epitopes in Cell Walls of Forage Chicory (Cichorium intybus) Leaves

The cell walls of forage chicory (Cichorium intybus) leaves are known to contain high proportions of pectic polysaccharides. However, little is known about the distribution of pectic polysaacharides among walls of different cell types/tissues and within walls. In this study, immunolabelling with four monoclonal antibodies was used to map the distribution of pectic polysaccharides in the cell walls of the laminae and midribs of these leaves. The antibodies JIM5 and JIM7 are specific for partially methyl-esterified homogalacturonans; LM5 and LM6 are specific for (1→4)-β-galactan and (1→5)-α-arabinan side chains, respectively, of rhamnogalacturonan I. All four antibodies labelled the walls of the epidermal cells with different intensities. JIM5 and JIM7, but not LM5 or LM6, labelled the middle lamella, tricellular junctions, and the corners of intercellular spaces of ground, xylem and phloem parenchyma. LM5, but not LM6, strongly labelled the walls of the few sclerenchyma fibres in the phloem of the midrib and lamina vascular bundles. The LM5 epitope was absent from some phloem parenchyma cells. LM6, but not LM5, strongly labelled the walls of the stomatal guard cells. The differential distribution of pectic epitopes among walls of different cell types and within walls may reflect the deposition and modification of these polysaccharides which are involved in cell wall properties and cell development.

Physical, Anatomical, and Biochemical Composition of Skins Cell Walls from Two Grapevine Cultivars (Vitis vinifera) of Champagne Region Related to Their Susceptibility to Botrytis cinerea during Ripening

This work investigated the structural, biochemical, and molecular characteristics of grape skin cell wall during ripening, related to susceptibility to Botrytis cinerea. The comparative study between the two main grape cultivars in Champagne region, Pinot noir and Chardonnay, quantified: (1) the maturity and physical profile of grape skin; (2) the morphological characteristics; (3) soluble pectic polysaccharides located in grape skin cell walls; and (4) the gene expression of the two main degrading enzymes (VvPME1 and VvPG1) and PME activity. During the maturation period, the grape skins of the two cultivars appear different in their structure and composition. Chardonnay is characterized by higher relative humidity (RH) and level of VvPG1 expression, lower disease incidence and penetrometry values, and thicker cell walls than Pinot noir skins. Thus, the cell wall composition is sufficiently different between grape varieties from the same area to allow their discrimination and could be used to better manage the harvest date.