Pectin forms polymeric pigments by complexing anthocyanins during red winemaking and ageing.

The long-term stability of red wine color depends on the formation of polymeric pigments from anthocyanins. Although there is still a lot of uncertainty about the specific structure of this diverse group of pigments, there is consensus that they are reaction products of anthocyanins and other polyphenols. Interactions between anthocyanins and pectic polysaccharides have been suggested to stabilize anthocyanins. This study explores the impact of such interactions by adding pectin during red winemaking. The results demonstrate that these interactions induce the formation of additional polymeric pigments which enhance the pigment stability during fermentation and aging. While initial pigment formation is higher in wines with added pectin, a notable proportion of the complexes degrades in the later stages of fermentation. Presumably, tannins form insoluble complexes with pectin, reducing tannin concentration by more than 300 mg/L. Anthocyanin concentrations decrease by over 400 mg/L, and polymeric pigments double. Anthocyanins that form polymeric pigments with pectic polysaccharides expand the range of pigments in red wines with possible consequences for the sensory properties of the wine. These findings highlight the complex interactions between pectin, anthocyanins, and tannins, and their influence on pigment formation and wine composition during fermentation and aging.

Improved Generation, Physicochemical Characteristics, and Food Application Studies of a Red Colorant Obtained from Oxidative Coupling of Chlorogenic Acid and Tryptophan.

Due to a widespread consumer reluctance toward synthetic food dyes, the interest in natural compounds from plants has increased. This study aimed to optimize the oxidation process between chlorogenic acid (CQA) and tryptophan (Trp) using sodium periodate (NaIO4) to obtain a red-colored pigment. The impact of temperature and different ratios of Trp to NaIO4 on the reaction progress was investigated. After the best conditions for the reaction were established, three pH values were tested. The reaction time could be reduced from 72 to 24 h with a yield of 46 ± 2% w/w based on the quantity of CQA. After the first purification step of the product by size exclusion chromatography, the pigment obtained was characterized for its solubility, and its hydrolyzed form was used for investigations into the stability at different pH values, storage under light and in the dark (period of 28 days), in the presence of reducing agents, and for heat resistance. Finally, several food matrices were successfully colored with the natural pigment in amounts from 0.005 to 0.01% (w/w). In conclusion, the present study provides new insights into the feasible production and comprehensive characterization of a red pigment derived from oxidative coupling of CQA and Trp, as well as its application in food systems.

Toward gentle chokeberry juice production by ultrasound-assisted enzymatic maceration.

Sustainable processes accompanied by high extraction yields and minimized amounts of by-products are a major goal of current fruit juice production. Controlled degradation of cell wall polysaccharides, in particular pectin, may contribute to reduced emergence of side streams. Possible strategies for the optimization are the selection of enzyme preparations based on comprehensive studies of their activities, the adjustment of maceration temperature toward more gentle conditions, and the application of alternative technologies such as ultrasound (US) during maceration. The present study provides insights into the effects of ultrasound-assisted enzymatic maceration (UAEM) on pectin degradation, total anthocyanin content, thermal and storage stability, and juice yield during chokeberry juice production on pilot-plant scale. The two enzyme preparations applied predominantly possessed polygalacturonase or pectin lyase activity. Cell wall polysaccharide degradation was improved by US and resulted in a 3% increase in juice yield by UAEM using an enzyme preparation that shows mostly polygalacturonase activity. Thermostability of anthocyanins was improved in juices produced using pectin lyase and applying US and matched the stability of anthocyanins in juices produced using polygalacturonase. Storage stability of anthocyanins was improved in juice produced using polygalacturonase during UAEM. UAEM also resulted in lower yields of pomace making the production more resource-efficient. Overall, the use of polygalacturonase has promising potential to advance conventional chokeberry juice production by applying US at gentle conditions.

Generation and structure elucidation of a red colorant formed by oxidative coupling of chlorogenic acid and tryptophan.

In view of the poor acceptance of synthetic food colorants by consumers, there is intense interest in novel natural compounds, preferably from plant-derived sources. We oxidized chlorogenic acid using NaIO4 and reacted the resultant quinone with tryptophan (Trp) to obtain a red-colored product. The colorant was precipitated, freeze-dried, purified by size exclusion chromatography, and subsequently characterized using UHPLC-MS, high-resolution mass spectrometry, and NMR spectroscopy. Additional mass spectrometric studies were performed on the reaction product generated with Trp educts labeled with 15N and 13C. The data obtained from these studies allowed the identification of a complex compound consisting of two Trp and one caffeic acid moieties, and the proposition of a tentative pathway of its formation. Thus, the present investigation expands our knowledge about the formation of red colorants based on the reaction of plant phenols and amino acids.

Preparative Fractionation of Phenolic Compounds and Isolation of an Enriched Flavonol Fraction from Winemaking Industry By-Products by High-Performance Counter-Current Chromatography.

High-performance counter-current chromatography (HPCCC) was used as a tool for the isolation and fractionation of phenolic compounds (PCs) in extracts from wine lees (WL) and grape pomace (GP). The biphasic solvent systems applied for HPCCC separation were n-butanol:methyl tert-butyl ether:acetonitrile:water (3:1:1:5) with 0.1% trifluoroacetic acid (TFA) and n-hexane:ethyl acetate:methanol:water (1:5:1:5). After refining the ethanol:water extracts of GP and WL by-products by ethyl acetate extraction, the latter system yielded an enriched fraction of the minor family of flavonols. Recoveries of 112.9 and 105.9 mg of purified flavonols (myricetin, quercetin, isorhamnetin, and kaempferol) in GP and WL, respectively, from 500 mg of ethyl acetate extract (equivalent to 10 g of by-product) were obtained. The HPCCC fractionation and concentration capabilities were also exploited for the characterization and tentative identification of constitutive PCs by ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS). In addition to the isolation of the enriched flavonol fraction, a total of 57 PCs in both matrixes were identified, 12 of which were reported for the first time in WL and/or GP. The application of HPCCC to GP and WL extracts may be a powerful approach to isolate large amounts of minor PCs. The composition of the isolated fraction demonstrated quantitative differences in the individual compound composition of GP and WL, supporting the potential exploitation of these matrixes as sources of specific flavonols for technological applications.

Grape-derived pectic polysaccharides alter the tannin and pigment composition of Cabernet Sauvignon red wines.

Tannins, anthocyanins, and polymeric pigments are essential phenolic constituents of red wine because they provide color, color stability, and mouthfeel properties like astringency. The behavior of these compounds is significantly affected by pectic polysaccharides, whereby the extent of their influence on red wine quality depends on their structural features and their interactions with the polyphenols. In the present study, the composition of the pectic polysaccharides of commercially available Cabernet Sauvignon wines and their impact on anthocyanin, tannin, and polymeric pigment analyses was characterized. This was accomplished by preparation of polysaccharide deprived wines and comparison of the polyphenolic composition of both, the wines and their corresponding polysaccharide-free counterparts. The results show that the cell wall fragments enhance the spectral absorbance of anthocyanins by facilitating anthocyanin self-association, leading to a co-pigmentation-like effect. Low molecular weight pectins like rhamnogalacturonan II and polygalacturonic acids with a low degree of esterification are assumed to form soluble complexes with anthocyanins and also prevent protein precipitation of tannins, which was reduced by 6-13%. High molecular weight pectins with a high degree of esterification lead to the increased precipitability of pigments and tannins by a factor of 1.3 to 32.4 and 1.1 to 1.9, respectively, seemingly impairing the incorporation of anthocyanins in tannins to form precipitable polymeric pigments that are responsible for the longevity of red wine color. The increased precipitability of the pigments due to the interactions with the polysaccharides may indicate the formation of pigmented yet non-covalent aggregates that show comparable properties to the covalently formed precipitable pigments. The formation of those non-covalent structures may affect red wine color stability and astringency.

Effects of flavonoids on membrane adaptation of food-associated bacteria.

The effects of naringenin and the biflavonoids amentoflavone and tetrahydroamentoflavone on select bacterial lipids (carotenoids, fatty acids, and menaquinones) and membrane fluidity based on Laurdan generalized polarization were investigated. For this purpose, the pigment-forming food-associated microorganisms Staphylococcus xylosus (DSM 20266T and J70), Staphylococcus carnosus DSM 20501T, and Micrococcus luteus (ATCC 9341 and J3) were studied. The results suggest an envelope stress response by microorganisms due to flavonoids and an employment of adaptive mechanisms using carotenoids, fatty acids, and menaquinones. The flavonoid monomer naringenin impacted carotenoids, fatty acids, menaquinones, and membrane fluidity. Naringenin significantly influenced the carotenoid profile, particularly by an increase in the relative proportion of 4,4'-diaponeurosporenoic acid in Staphylococcus xylosus. Amentoflavone caused changes mainly in the membrane of Micrococcus luteus and decreased the menaquinone content. Tetrahydroamentoflavone mainly affected the carotenoids in the investigated strains. The noticeably different CCS value of tetrahydroamentoflavone compared to naringenin and amentoflavone revealed further insights into the structure-dependent effects of flavonoids. This study provides valuable insights into the response of pigment-forming food-associated microorganisms to naringenin, amentoflavone, and tetrahydroamentoflavone, which is important for the targeted and safe application of the latter as natural preservatives and useful for further research on the mechanisms of action.

Conversion of hydroxycinnamic acids by Furfurilactobacillus milii in sorghum fermentations: Impact on profile of phenolic compounds in sorghum and on ecological fitness of Ff. milii.

The conversion of phenolic compounds by lactobacilli in food fermentations contributes to food quality. The metabolism of phenolics by lactobacilli has been elucidated in the past years but information on the contribution of specific enzymes in food fermentations remains scarce. This study aimed to address this gap by disruption of genes coding for the hydroxycimmanic acid reductase Par1, the hydroxycinnamic acid decarboxylase Pad, the hydrocinnamic esterase EstR, and strains with disruption of all three genes in Furfurilactobacillus milii FUA3583. The conversion of phenolics by Ff. milii and its isogenic mutants in sorghum fermentations was studied by LC-UV and LC-UV-MS/MS analyses. Ff. milii FUA3583 converted hydroxycinnamic acids predominantly with Par1. Vinylphenols were detected only in mutants lacking par1. A phenotype for the estR defective mutant was not identified. The formation of pyrano-3-deoxyanthocyanidins was observed only after fermentation with strains expressing Pad. Specifically, formation of these compounds was low with Ff. milii FUA3583, substantially increased in the Par1 mutant and abolished in all mutants with disrupted pad. Competition experiments with Ff. milii FUA3583 and its isogenic mutants demonstrated that expression of one of the two metabolic pathways for hydroxycinnamic acids increases the ecological fitness of the strain. Disruption of EstR in a Δpar1Δpar2Δpad background improved ecological fitness, indirectly demonstrating a phenotype of the esterase in Ff. milii. The documentation of the functionality of genes coding for conversion of hydroxycinnamic acids may support the selection of starter cultures for improved quality of fermented cereal products.

Recent Advances in Research on Polyphenols: Effects on Microbiota, Metabolism, and Health.

Polyphenols have attracted huge interest among researchers of various disciplines because of their numerous biological activities, such as antioxidative, antiinflammatory, antiapoptotic, cancer chemopreventive, anticarcinogenic, and antimicrobial properties, and their promising applications in many fields, mainly in the medical, cosmetics, dietary supplement and food industries. In this review, the latest scientific findings in the research on polyphenols interaction with the microbiome and mitochondria, their metabolism and health beneficial effects, their involvement in cognitive diseases and obesity development, as well as some innovations in their analysis, extraction methods, development of cosmetic formulations and functional food are summarized based on the papers presented at the 13th World Congress on Polyphenol Applications. Future implications of polyphenols in disease prevention and their strategic use as prophylactic measures are specifically addressed. Polyphenols may play a key role in our tomorrow´s food and nutrition to prevent many diseases.

Application of Crude Pomace Powder of Chokeberry, Bilberry, and Elderberry as a Coloring Foodstuff.

Berry pomace, rich in polyphenols, especially anthocyanins, accumulates during the production of red juices. Pomace from chokeberry (Aronia melanocarpa Michx.), bilberry (Vaccinium myrtillus L.), and elderberry (Sambucus nigra L.) represent good sources of coloring foodstuffs. Pomace powders (PP) were prepared by milling the seedless fractions of the three dried berry pomaces (50 °C, 8 h). Techno-functional properties of the powders such as particle size distribution, bulk density, sedimentation velocity, and swelling capacity were determined to evaluate the powders for possible food applications. Total anthocyanin content was quantified by UHPLC-DAD before and during a storage experiment to monitor the degradation of anthocyanins in the PP and in a yogurt model application. The high content of phenolic compounds and the still intact cell structure ensured high stability of anthocyanins over 28 days of storage. In the model application, color saturation was stable over the whole storage time of 14 days. Regarding the techno-functional properties, only a few differences between the three PP were observed. The particle size of elderberry PP was larger, resulting in lowest bulk density (0.45 g/mL), high cold-water solubility (16.42%), and a swelling capacity of 10.16 mL/g dw. Sedimentation velocity of the three PP was fast (0.02 mL/min) due to cluster formation of the particles caused by electrostatic and hydrophobic properties. Compared to other high-intensity coloring foodstuffs, the use of PP, showing acceptable color stability with potential health-promoting effects, represents a wide applicability in different food applications and especially in products with a longer shelf-life.

Effects of Extraction Conditions on Banana Peel Polyphenol Oxidase Activity and Insights into Inactivation Kinetics Using Thermal and Cold Plasma Treatment.

The objective of this work was to characterize banana peel polyphenol oxidase (PPO) and to study the inactivation kinetics during thermal and cold atmospheric pressure plasma treatment. Since varietal differences in enzyme characteristics are a well-known phenomenon, 'Prata' banana peel PPO was characterized, and PPO activity and thermal stability of the peel PPO of the two dessert banana cultivars 'Cavendish' and 'Prata' were compared to identify the cultivar better suited for industrial food applications. A crude extract obtained from the peels of the Brazilian banana variety 'Prata' revealed highest PPO activities (46.0-55.2 nkat/mL) at 30-40 °C in a range of pH 6.0-6.5 after addition of 0.5 g/gsample polyvinylpyrrolidone and 0.5% (v/v) Triton X-100 during extraction. 'Cavendish' PPO activity was four times higher. Banana peel PPO exhibited the highest affinity towards dopamine (KM = 0.94 mM). Thermal inactivation of 'Prata' and 'Cavendish' PPO was achieved at 90 °C after 5 and 15 min, respectively, whereas cold plasma treatment did not decrease PPO activity below 46% of the initial enzyme activity. The inactivation behavior of PPO could successfully be described by a two-fraction model indicating at least two types of isoenzymes with different thermal stability. The overall high thermal stability was mainly attributed to membrane-bound PPO. The results may help to prevent enzymatic browning of banana peels and thereby facilitate their valorization as food ingredients.

Methylation of Cyanidin-3-O-Glucoside with Dimethyl Carbonate.

The approach presented in this study is the first for the hemisynthesis of methylated anthocyanins. It was possible to obtain cyanidin-3-O-glucoside derivatives with different degrees of methylation. Cautious identification of 4'-, 5-, and 7-OH monomethylated derivatives was also accomplished. The methylation agent used was the "green chemical" dimethyl carbonate (DMC), which is characterized by low human and ecological toxicity. The influence of the temperature, reaction time, and amount of the required diazabicyclo[5.4.0]undec-7-en (DBU) catalyst on the formation of the products was examined. Compared to conventional synthesis methods for methylated flavonoids using DMC and DBU, the conditions identified in this study result in a reduction of reaction time, and an important side reaction, so-called carboxymethylation, was minimized by using higher amounts of catalyst.

Chemical Hemisynthesis of Sulfated Cyanidin-3-O-Glucoside and Cyanidin Metabolites.

The metabolism of anthocyanins in humans is still not fully understood, which is partly due to the lack of reference compounds. It is known that sulfation is one way of the complex phase II biotransformation mechanism. Therefore, cyanidin-3-O-glucoside and the cyanidin aglycone were chemically converted to their sulfates by reaction with sulfur trioxide-N-triethylamine complex in dimethylformamide. The reaction products were characterized by UHPLC coupled to linear ion trap and IMS-QTOF mass spectrometry. Based on MS data, retention times, and UV-Vis spectra, the compounds could tentatively be assigned to A-, C-, or B-ring sulfates. Analysis of urine samples from two volunteers after ingestion of commercial blackberry nectar demonstrated the presence of two sulfated derivatives of the cyanidin aglycone and one sulfated derivative of the cyanidin-3-O-glucoside. It was found that both the A ring and the B ring are sulfated by human enzymes. This study marks an important step toward a better understanding of anthocyanin metabolism.

Effects of ultrasound on the enzymatic degradation of pectin.

Ultrasound-assisted enzymatic maceration (UAEM) has gained considerable interest in the fruit juice industry, owing to its potential to increase juice yield and content of polyphenols while simultaneously saving time and energy. In this study, the effects of UAEM (ultrasonic probe, 20 kHz, 21 W*cm-2 and 33 W*cm-2) on pectin degradation in a continuous circulation system were investigated over 60 and 90 min. Main pectinolytic enzymes activities of (polygalacturonase, pectin lyase and pectin methylesterase) of a commercial enzyme preparation were examined for individual synergistic effects with US. Pectin hydrolysis by UAEM differed significantly compared to treatment with ultrasound or enzymes alone regarding the profile of degradation products compared to treatment with ultrasound or enzymes alone. Ultrasound fragmented pectin to less branched oligomers of medium molecular weight (Mp approx. 150 kDa), which were further degraded by pectinolytic activities. The low molecular weight fraction (<30 kDa), which is known to be beneficial for juice-quality by adding nutritional value and stabilizing polyphenols, was enriched in small oligomers of homogalacturonan-derived, rhamnogalacturonan I-derived, and rhamnogalacturonan II-derived residues. Synergistic effects of ultrasound application enhanced the effective activities of polygalacturonase and pectin lyase and even prolonged their performance over 90 min, whereas the effective activity of pectin methylesterase was not affected. Final marker concentrations determined by each enzyme assay revealed a considerable higher total process output after UAEM treatment at reduced temperature (30 °C) comparable to the output after conventional batch maceration at 50 °C. The obtained results demonstrate the high potential of UAEM to produce high-quality juice by controlling pectin degradation while reducing process temperature and equally highlight the matrix and enzyme specific effects of a simultaneous US treatment.

Volatile Phenols-Important Contributors to the Aroma of Plant-Derived Foods.

Volatile phenols like phenylpropanoid and benzoid compounds originate from the aromatic amino acid phenylalanine, which is biosynthesized via the shikimate/arogenate pathway. These volatile compounds contribute to the aroma of a number of economically important plant-derived foods like herbs, spices and fruits. The sequestration of numerous phenylpropanoid and benzoid compounds as glycosides occurs widely in fruits, and this pool represents an important source of flavor that can be released during storage and processing. Therefore, this review will provide an overview of the biosynthesis of free and glycosylated phenylpropanoid and benzoid compounds and their reactions during food processing, which both lead to the generation of odor-active volatile phenols in plant-derived foods.

Effects of carrier agents on powder properties, stability of carotenoids, and encapsulation efficiency of goldenberry (Physalis peruviana L.) powder produced by co-current spray drying.

Maltodextrin, modified starch, inulin, alginate, gum arabic, and combinations thereof were used as carrier agents for spray drying of carotenoid-rich goldenberry (Physalis peruviana L.) juice and compared to cellobiose as an alternative carrier. Powders were analyzed with respect to particle size and morphology, yield, moisture content, cold water solubility, suspension stability, hygroscopicity, carotenoid encapsulation efficiency, and carotenoid retention during storage. A high initial carotenoid concentration after spray drying, a high encapsulation efficiency of 77.2%, and a slow carotenoid degradation kinetics favored the high carotenoid content of the cellobiose powder at the end of the storage. Cellobiose might protect the carotenoids from degradation processes by light exposure, high temperature, and oxygen due to a tighter particle crust and larger particle sizes. Therefore, cellobiose may be considered a potential carrier agent for the encapsulation of carotenoid-rich fruit juices.

Impact of Different Pasteurization Techniques and Subsequent Ultrasonication on the In Vitro Bioaccessibility of Carotenoids in Valencia Orange (Citrus sinensis (L.) Osbeck) Juice.

The effects of traditional pasteurization (low pasteurization, conventional pasteurization, hot filling) and alternative pasteurization (pulsed electric fields, high pressure processing), followed by ultrasonication on the carotenoid content, carotenoid profile, and on the in vitro carotenoid bioaccessibility of orange juice were investigated. There was no significant difference in the total carotenoid content between the untreated juice (879.74 µg/100 g juice) and all pasteurized juices. Significantly lower contents of violaxanthin esters were found in the high thermally-treated juices (conventional pasteurization, hot filling) compared to the untreated juice, owing to heat-induced epoxy-furanoid rearrangement. The additional ultrasonication had almost no effects on the carotenoid content and profile of the orange juices. However, the in vitro solubilization and the micellarization efficiency were strongly increased by ultrasound, the latter by approximately 85.3-159.5%. Therefore, among the applied processing techniques, ultrasonication might be a promising technology to enhance the in vitro bioaccessibility of carotenoids and, thus, the nutritional value of orange juice.

Effects of structural differences on the antibacterial activity of biflavonoids from fruits of the Brazilian peppertree (Schinus terebinthifolius Raddi).

Flavonoids, synthesized by plants across all families and therefore found in a huge variety, possess a diverse range of pharmacological properties. Direct antibacterial and synergistic activities as well as the inhibition of several bacterial virulence factors are known. Besides the mode of action, it is important to understand the structure-activity relationship to identify key structural characteristics. This study aimed to identify biflavonoids with antibacterial activity from Schinus terebinthifolius Raddi fruits. The purified biflavonoids were characterized in terms of their antibacterial effects. We found that the activity of biflavonoids, including agathisflavone (AGF), amentoflavone (AMF), and tetrahydroamentoflavone (THAF), was dependent on their chemical configuration and degree of oxidation, with THAF showing the highest activity on planktonic cells. Additionally, biofilm formation and composition were strongly influenced by THAF. Even slight differences in the chemical structure have fundamental effects on the activity of isolated biflavonoids. This suggests a specific binding of these substances in bacteria and thus enables detailed investigations of the mode of action in the future.

Enhancement of Biomimetic Enzymatic Mineralization of Gellan Gum Polysaccharide Hydrogels by Plant-Derived Gallotannins.

Mineralization of hydrogel biomaterials with calcium phosphate (CaP) is considered advantageous for bone regeneration. Mineralization can be both induced by the enzyme alkaline phosphatase (ALP) and promoted by calcium-binding biomolecules, such as plant-derived polyphenols. In this study, ALP-loaded gellan gum (GG) hydrogels were enriched with gallotannins, a subclass of polyphenols. Five preparations were compared, namely three tannic acids of differing molecular weight (MW), pentagalloyl glucose (PGG), and a gallotannin-rich extract from mango kernel (Mangifera indica L.). Certain gallotannin preparations promoted mineralization to a greater degree than others. The various gallotannin preparations bound differently to ALP and influenced the size of aggregates of ALP, which may be related to ability to promote mineralization. Human osteoblast-like Saos-2 cells grew in eluate from mineralized hydrogels. Gallotannin incorporation impeded cell growth on hydrogels and did not impart antibacterial activity. In conclusion, gallotannin incorporation aided mineralization but reduced cytocompatibility.