Characterization, antioxidant and antitumor activities of phenolic compounds from Amomum villosum Lour.

Amomum villosum Lour. (A. villosum), known as Sharen in China, is widely used for culinary and medicinal purposes due to containing a diverse set of bioactive compounds. In this study, the optimum ethanol extraction process was optimized and the composition and biological activities (antioxidant and antitumor) of five different fractions (dichloromethane, petroleum ether, ethyl acetate, n-butanol and H2O) extracted from the ethanol extract of A. villosum were investigated. The results showed that the optimal extraction conditions were extraction temperature 80°C, extraction time 120 min, ethanol concentration 40% and solid-liquid ratio 1:25 g/mL. Moreover, 35 bioactive compounds were successfully identified by UPLC-ESI-QTOF-MS/MS from five factions for the first time, including 12 phenolic acids and derivatives, 2 organic acids, 12 flavonoids and derivatives, 2 oxylipins and 7 proanthocyanidins. Among them, ethyl acetate fraction (Fr-EtOAc) exhibited the highest content of total phenolic (374.01 mg GAE/g DW) and flavonoid (93.11 mg RE/g DW), where vanillic acid, catechin, epicatechin and protocatechuic acid were the predominant phenolic compounds that accounting for 81.65% of the quantified bioactive compounds. In addition, Fr-EtOAc demonstrated excellent total antioxidant activity (IC50 of DPPH and ABTS assays were 0.23, 0.08 mg/mL, respectively, and FRAP assay was 322.91 mg VCE/100 g DW) and antitumor activity (1,000 µg/mL, 79.04% inhibition rate). The results could provide guidance for the industrial production and application of A. villosum.

Phycocyanin-chitosan complex stabilized emulsion: Preparation, characteristics, digestibility, and stability.

Phycocyanin is a natural pigment protein with antioxidant, anti-tumor, and anti-inflammatory properties, but its relatively poor emulsibility limits its use in the food industry. In order to improve the emulsifying capacity of phycocyanin, a novel phycocyanin-chitosan complex was prepared, and the characteristics, digestibility, and stability of emulsion containing oil droplets stabilized by the complex were investigated. The results showed that the phycocyanin-chitosan complex had better stability and lower interfacial tension at pH 6.5 than phycocyanin, and it significantly improved the stability of emulsion and inhibited the aggregation of oil droplets. The phycocyanin-chitosan complex stabilized emulsion showed better physical stability, digestibility, and oxidation stability than the phycocyanin emulsion. The particle size of the phycocyanin-chitosan complex stabilized emulsion was very small (from 0.1 to 2 µm), and its absolute value of zeta potential was high. Overall, this study suggests that the phycocyanin-chitosan complex effectively improved the emulsifying capacity of phycocyanin.

Investigation of peanut allergen-procyanidin non-covalent interactions: Impact on protein structure and in vitro allergenicity.

Interactions between plant polyphenols and food allergens may be a new way to alleviate food allergies. The non-covalent interactions between the major allergen from peanut (Ara h 2) with procyanidin dimer (PA2) were therefore characterized using spectroscopic, thermodynamic, and molecular simulation analyses. The main interaction between the Ara h 2 and PA2 was hydrogen bonding. PA2 statically quenched the intrinsic fluorescence intensity and altered the conformation of the Ara h 2, leading to a more disordered polypeptide structure with a lower surface hydrophobicity. In addition, the in vitro allergenicity of the Ara h 2-PA2 complex was investigated using enzyme-linked immunosorbent assay (ELISA) kits. The immunoglobulin E (IgE) binding capacity of Ara h 2, as well as the release of allergenic cytokines, decreased after interacting with PA2. When the ratio of Ara h 2-to-PA2 was 1:50, the IgE binding capacity was reduced by around 43 %. This study provides valuable insights into the non-covalent interactions between Ara h 2 and PA2, as well as the potential mechanism of action of the anti-allergic reaction caused by binding of the polyphenols to the allergens.

Macadamia oil-based oleogels as cocoa butter alternatives: Physical properties, oxidative stability, lipolysis, and application.

In this study, macadamia oil-based oleogels were prepared using monoglyceride stearate (MG) as a gelator with a low critical gelation concentration (3.0 wt%). The physical properties of the oleogels were evaluated by polarized light microscopy, Fourier transform infrared spectroscopy, differential scanning calorimetry, X-ray diffraction, texture and rheological analysis. And the lipid digestion and oxidative stability of the macadamia oil were determined by pH titration and accelerated oxidation test, respectively. The results showed that the hardness, oil binding capacity, and thermal stability of the oleogels increased with increasing MG concentration, which was attributed to the formation of a network of MG crystals held together by van der Waals interactions and hydrogen bonds. Rheological analysis indicated that all the oleogels exhibited a thermally reversible solid-to-liquid transition and viscoelastic behavior at ambient temperatures. Moreover, the formation of oleogels increased fatty acid release during in vitro lipid digestion and improved the oxidative stability of the macadamia oil. In addition, the potential application of these oleogels as replacements for saturated fats in foods was demonstrated by creating a chocolate product where the cocoa butter was replaced with macadamia oil-based oleogels with a high degree of unsaturation. These results can provide guidance for the preparation of macadamia oil-based oleogels, which may increase their application in foods.

The interaction and physicochemical properties of the starch-polyphenol complex: Polymeric proanthocyanidins and maize starch with different amylose/amylopectin ratios.

This study investigated the impact of polymeric proanthocyanidins (PPC) on the physicochemical characteristics of maize starch with varying amylose content, and their potential interaction mechanism. PPC with a lower content (1 %) reduced the viscoelasticity of the high amylose maize starch (HAM) system, inhibited amylose rearrangement, and enhanced its fluidity. However, excessive PPC restrained the interaction between PPC and amylose. In contrast to HAM, PPC improved the gelation ability of waxy maize starch (WAM) as PPC concentration was raised. PPC suppressed the recrystallization of starch during storage, and PPC had a superior inhibition influence on the retrogradation of WAM in comparison to HAM. This indicated that amylopectin was more likely to interact with PPC than amylose. Hydrogen bonds were the main driving force between PPC and starch chains, which was clarified by Fourier transform-infrared, nuclear magnetic resonance, X-ray diffraction, iodine bonding reaction, and dynamic light scattering data. Additionally, the mechanism of interaction between PPC and the two starch components may be similar, and variance in physicochemical attributes can be primarily credited to the percentage of amylose to amylopectin in starch.

Investigation of bovine ß-lactoglobulin-procyanidin complexes interactions and its utilization in O/W emulsion.

Procyanidins are bioactive polyphenols that have a strong affinity to proteins. Beta-lactoglobulin (BLG) is widely used as an emulsifier in the food and other industries. This study evaluated the interaction between BLG and A-type procyanidin dimer (PA2) using the spectroscopic, thermodynamic, and molecular simulation. PA2 decreased the transmissivity and quenched the intrinsic fluorescence of BLG, suggesting that the two molecules formed a complex. The binding of PA2 reduced the surface hydrophobicity and altered the conformation of BLG with increasing the random coil regions. Thermodynamic and isothermal titration calorimetry analyses suggested that the main driving force of PA2-BLG interaction was hydrophobic attraction. Molecular docking simulations were used to identify the main interaction sites and forces in the BLG-PA2 complexes, which again indicated that hydrophobic interactions dominated. In addition, the influence of PA2 on the ability of BLG to form and stabilize O/W emulsions was analyzed. Emulsions formulated using BLG-PA2 complexes contained relatively small droplets (D4,3 ≈ 0.7 µm) and high surface potentials (absolute value >50 mV). Compared to BLG alone, BLG-PA2 complexes improved the storage stability of the emulsions. This study provides valuable new insights into the formation, properties, and application of protein-polyphenol complexes as functional ingredients in foods.

Hypolipidemic effects of macadamia oil are related to AMPK activation and oxidative stress relief: In vitro and in vivo studies.

Macadamia oil is rich in monounsaturated fatty acids, especially a high level of palmitoleic acid, which may have beneficial health effects by lowering blood lipid levels. In our study, the hypolipidemic effects of macadamia oil and its potential mechanisms of action were investigated using a combination of in vitro and in vivo assays. The results showed that macadamia oil significantly reduced lipid accumulation, and improved triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) levels in oleic acid-induced high-fat HepG2 cells. The macadamia oil treatment also exhibited antioxidant effects, as seen by its ability to reduce reactive oxygen species and malondialdehyde (MDA) levels, and increase superoxide dismutase (SOD) activity. The effects of 1000 µg/mL of macadamia oil were comparable to that of 4.19 µg/mL simvastatin. The results of qRT-PCR and western blotting analyses indicated that macadamia oil effectively inhibited hyperlipidemia by reducing the expression levels of SREBP-1c, PPAR-γ, ACC and FAS and by enhancing the expression levels of HO-1, NRF2 and γ-GCS, via AMPK activation and oxidative stress relief, respectively. In addition, different doses of macadamia oil were found to significantly improve liver lipid accumulation, reduce serum and liver TC, TG, and LDL-C levels, increase HDL-C levels, increase antioxidant enzyme (SOD, GSH-Px, and T-AOC) activity, and decrease the MDA content of mice on a high-fat diet. These results indicated that macadamia oil had a hypolipidemic effect and provide insights that might facilitate the development of functional food and dietary supplements.

Effects of cell wall polysaccharides on the bioaccessibility of carotenoids, polyphenols, and minerals: an overview.

Carotenoids, polyphenols, and minerals (CPMs) are representative bioactive compounds and micronutrients in plant-based foods, showing many potentially positive bioactivities. Bioaccessibility is a prerequisite for bioactivities of CPMs. Cell wall polysaccharides (CWPs) are major structural components of plant cell wall, and they have been proven to affect the bioaccessibility of CPMs in different ways. This review summarizes recent literatures about the effects of CWPs on the bioaccessibility of CPMs and discusses the potential mechanisms. Based on the current findings, CWPs can inhibit the bioaccessibility of CPMs in gastrointestinal tract. The effects of CWPs on the bioaccessibility of polyphenols and minerals mainly attributes to bind between them, while CWPs affect the bioaccessibility of carotenoids by changing the digestive environment. Further, this review overviews the factors (environmental conditions, CWPs properties and CPMs characteristics) affecting the interactions between CWPs and CWPs. This review may help to better design healthy and nutritious foods precisely.

Improving foam performance using colloidal protein-polyphenol complexes: Lactoferrin and tannic acid.

In this study, colloidal complexes were prepared from bovine lactoferrin (BLF) and tannic acid (TA) and then their ability to form and stabilize foams was characterized. The molecular interactions between BLF and TA were studied using fluorescence and molecular docking analysis, which suggested that hydrophobic forces were primarily involved in holding the complexes together. The production of colloidal BLF-TA complexes was supported by increases in turbidity and mean particle diameter, quenching of intrinsic fluorescence, decrease in surface hydrophobicity, and change in conformation. When used alone, BLF exhibited good foam formation but poor foam stability properties. In contrast, BLF-TA complexes exhibited good foam stability but poor foamability properties. The change in foaming properties of the proteins was closely related to their interactions with the polyphenols. These findings may be useful for the development of novel functional ingredients to construct food foams with good physicochemical and nutritional attributes.

Comparative Study of Chemical Compositions and Antioxidant Capacities of Oils Obtained from 15 Macadamia (Macadamia integrifolia) Cultivars in China.

The planting area of macadamia in China accounted for more than one third of the world's planted area. The lipid compositions, minor components, and antioxidant capacities of fifteen varieties of macadamia oil (MO) in China were comparatively investigated. All varieties of MO were rich in monounsaturated fatty acids, mainly including oleic acid (61.74-66.47%) and palmitoleic acid (13.22-17.63%). The main triacylglycerols of MO were first time reported, including 19.2-26.1% of triolein, 16.4-18.2% of 1-palmitoyl-2,3-dioleoyl-glycerol, and 11.9-13.7% of 1-palmitoleoyl-2-oleoyl-3-stearoyl-glycerol, etc. The polyphenol, α-tocotrienol and squalene content varied among the cultivars, while Fuji (791) contained the highest polyphenols and squalene content. Multiple linear regression analysis indicated the polyphenols and squalene content positively correlated with the antioxidant capacity. This study can provide a crucial directive for the breeding of macadamia and offer an insight into industrial application of MO in China.

Fabrication of Oil-in-Water Emulsions with Whey Protein Isolate-Puerarin Composites: Environmental Stability and Interfacial Behavior.

Protein-polyphenol interactions influence emulsifying properties in both directions. Puerarin (PUE) is an isoflavone that can promote the formation of heat-set gels with whey protein isolate (WPI) through hydrogen bonding. We examined whether PUE improves the emulsifying properties of WPI and the stabilities of the emulsions. We found that forming composites with PUE improves the emulsifying properties of WPI in a concentration-dependent manner. The optimal concentration is 0.5%, which is the highest PUE concentration that can be solubilized in water. The PUE not only decreased the droplet size of the emulsions, but also increased the surface charge by forming composites with the WPI. A 21 day storage test also showed that the maximum PUE concentration improved the emulsion stability the most. A PUE concentration of 0.5% improved the stability of the WPI emulsions against environmental stress, especially thermal treatment. Surface protein loads indicated more protein was adsorbed to the oil droplets, resulting in less interfacial WPI concentration due to an increase in specific surface areas. The use of PUE also decreased the interfacial tension of WPI at the oil-water interface. To conclude, PUE improves the emulsifying activity, storage, and environmental stability of WPI emulsions. This result might be related to the decreased interfacial tension of WPI-PUE composites.

Inhibitory mechanism of salicylic acid on polyphenol oxidase: A cooperation between acidification and binding effects.

Salicylic acid is generally considered to combine with polyphenol oxidase (PPO) to inhibit activity and enzymatic browning, while its acidification effect on PPO activity was usually neglected. In this study, the inhibitory mechanism of salicylic acid on PPO was examined from acidification and binding effects by altering the buffer conditions. As the buffer concentration increased, contribution of acidification decreased while the binding effect became more predominant. Salicylic acid exhibited competitive inhibition on PPO, inducing the changes in secondary structure with a reduction in α-helix. Molecular docking results showed that salicylic acid interacted with residues HIS61, HIS85, HIS259, HIS263 and VAL283 through hydrogen bond and hydrophobic interaction. Furthermore, acidic pH enhanced the binding of salicylic acid to PPO with lower binding energy, additional hydrogen bond and electrostatic interactions. Therefore, both acidification and binding effects were important for salicylic acid on PPO inhibition and enzymatic browning control in fruit and vegetables.

Effects of proanthocyanidins on the pasting, rheological and retrogradation properties of potato starch.

BACKGROUND: Proanthocyanidins (PAS) were complexed with potato starch (PS) to prepare polyphenol-starch complexes. The pasting, rheological and retrogradation properties of the complexes were investigated. RESULTS: The addition of PAS markedly affected the pasting, rheological and retrogradation properties of PS, especially at a concentration of 5% (w/w). Rapid viscosity analysis indicated that PAS significantly changed the viscosity, breakdown and setback value of PS. The rheological results showed that PAS decreased the flow behavior index and consistency coefficient, but increased the viscoelasticity of PS. Differential scanning calorimetry and X-ray diffraction indicated that PAS delayed the retrogradation of PS. Furthermore, scanning electron microscopy indicated that the morphologies of retrograded PS gels were greatly altered to a less compact structure with the presence of PAS. Moreover, Fourier transform infrared spectroscopy elucidated that PAS interacted with PS via a noncovalent interaction, and inhibited the retrogradation of PS. CONCLUSIONS: The findings suggested that supplementing PS with PAS might be an effective and convenient method for modifying the physicochemical properties of PS. © 2021 Society of Chemical Industry.

Protein-polyphenol functional ingredients: The foaming properties of lactoferrin are enhanced by forming complexes with procyanidin.

The solution turbidity and intrinsic fluorescence quenching increased after procyanidin was mixed with lactoferrin. The addition of procyanidin also caused a reduction in the surface hydrophobicity of the lactoferrin, suggesting procyanidin bound to non-polar patches on lactoferrin's surfaces. Moreover, the binding interaction caused an appreciable alteration in the structure of both the polyphenol and protein. Thermodynamic analysis indicated the interaction was spontaneous and mainly driven by entropy changes, suggesting that hydrophobic interactions dominated. A computational docking simulation provided insights into the location of the most-likely binding sites on the protein, as well as the nature of the interaction forces involved. In particular, both hydrophobic and hydrogen bonding were found to be important. The binding of the procyanidin to the lactoferrin enhanced its foaming properties. These results may lead to the development of a new class of natural functional ingredients that can be used in food products to improve their quality attributes.

Impact of pH, ferrous ions, and tannic acid on lipid oxidation in plant-based emulsions containing saponin-coated flaxseed oil droplets.

The influence of pH (pH 3, 5 and 7), ferrous ions (0 or 100 µM Fe2+), and tannic acid (0 to 0.1% TA) on the rate of lipid oxidation in plant-based emulsions containing quillaja saponin-coated flaxseed oil droplets was studied. Tannic acid formed complexes with Fe2+ whose properties depended on TA:Fe2+ ratio and pH. Emulsions were incubated at 37 °C in the dark, and changes in their particle size, surface potential, appearance, microstructure, and lipid oxidation status were monitored over time. The initial ζ-potential and mean particle diameter of the emulsions were -68 mV and 0.18 µm, respectively. In the absence of TA, the particle size increased appreciably during storage due to droplet coalescence, as rapid oxidation occurred. In the presence of TA, the emulsions were more resistant to both droplet aggregation and lipid oxidation, as a result of its strong ferrous ion-binding properties. The lipid oxidation rate increased with decreasing pH, which was attributed to an increase in ferrous ion's water-solubility and activity in acidic solutions. The addition of Fe2+ greatly accelerated lipid oxidation, but the oxidation rate was decreased by also adding TA. These results suggest that tannic acid is an effective antioxidant in emulsions, which can be attributed to its ferrous ion-chelation properties.

Comparison of Emulsion and Nanoemulsion Delivery Systems: The Chemical Stability of Curcumin Decreases as Oil Droplet Size Decreases.

The water dispersibility, chemical stability, and bioaccessibility of curcumin, a labile hydrophobic nutraceutical, can be enhanced by incorporating it inside the oil droplets of oil-in-water emulsions or nanoemulsions. In these multiphase systems, the curcumin remains relatively stable to degradation when surrounded by oil but degrades rapidly when surrounded by water. We hypothesized that the size of the lipid droplets would therefore impact the stability of encapsulated curcumin by altering the surface area of oil exposed to water. The effect of droplet surface area on the kinetics of curcumin degradation was therefore studied by producing emulsions with different mean droplet diameters (d32) and therefore different specific surface areas (AS): large (d32 = 20.9 µm; AS = 300 m2 kg-1); medium (d32 = 2.53 µm; AS = 2500 m2 kg-1); small (d32 = 0.26 µm; AS = 24,000 m2 kg-1); and very small (d32 = 0.083 µm; AS = 80,000 m2 kg-1) emulsions. All the emulsions initially had milky-yellow appearances and were relatively stable to aggregation during the course of the experiments. However, rapid creaming was observed in the large and medium emulsions because of their relatively large droplet size. The emulsions all exhibited some color fading during storage, with the rate of curcumin degradation increasing with decreasing droplet size. For instance, the percentage of curcumin remaining in the emulsions after 17 days storage was 91.4 ± 1.5 > 77.3 ± 6.6 > 66.7 ± 1.9 ≫ 30.6 ± 2.8% for the large, medium, small, and very small emulsions, respectively. The more rapid chemical degradation of the curcumin in the smaller droplets can be attributed to the fact that curcumin exchange between the interior and exterior of the droplets occurs more rapidly as the droplet dimensions decrease. Our results indicate that the droplet size plays a critical role in the degradation of curcumin encapsulated in emulsions, which may have important consequences for the formulation of curcumin-enriched foods and beverages with enhanced bioactivity. In particular, it suggests that emulsions are more effective at chemically stabilizing curcumin than nanoemulsions.

Utilization of plant-based protein-polyphenol complexes to form and stabilize emulsions: Pea proteins and grape seed proanthocyanidins.

Plant-based proteins and polyphenols are increasingly being explored as functional food ingredients. Colloidal complexes were prepared from pea protein (PP) and grape seed proanthocyanidin (GSP) and the ability of the PP/GSP complexes to form and stabilize oil-in-water emulsions were investigated. The main interactions between PP and GSP were hydrogen bonding. The stability of PP-GSP complexes to environmental changes were studied: pH (2-9); ion strength (0-0.3 M); and temperature (30-90 °C). Emulsions produced using PP-GSP complexes as emulsifiers had small mean droplet diameters (~200 nm) and strongly negative surface potentials (~-60 mV). Compared to PP alone, PP-GSP complexes slightly decreased the isoelectric point, thermostability, and salt stability of the emulsions, but increased their storage stability. The presence of GSP gave the emulsions a strong salmon (red-yellow) color, which may be beneficial for some specific applications. These results may assist in the creation of more efficacious food-based strategies for delivering proanthocyanidins.

Binding mechanism and antioxidant capacity of selected phenolic acid - ß-casein complexes.

Phenolic acids are added to some dairy products as functional ingredients. The molecular interactions between the phenolic acids and milk proteins impacts their functional performance and product quality. In this study, the interactions between a milk protein (ß-casein) and a number of phenolic acids was investigated: 3,4-dihydroxybenzoic acid (DA); gallic acid (GA); syringic acid (SA); caffeic acid (CaA); ferulic acid (FA); and, chlorogenic acid (ChA). The structural characteristics of the phenolic acids, such as type, hydroxylation, methylation, and steric hindrance, affected their binding affinity to ß-casein. The strength of the binding constant decreased in the following order: CaA > ChA > FA > SA > GA > DA. Cinnamic acid derivatives (CaA, FA, and ChA) exhibited a stronger binding affinity with ß-casein than benzoic acid derivatives (DA, GA, and SA). Hydrophobic forces and electrostatic interactions dominated the interactions of ß-casein with benzoic acid and cinnamic acid derivatives, respectively. The number of hydroxyl groups on the phenolic acids enhanced their binding ability, while steric hindrance effects reduced their binding ability. The influence of methylation depended on phenolic acid type. After binding with phenolic acids, the conformation of the ß-casein changed, with a loss of random coil structure, an increase in α-helix structure, and a decrease in surface hydrophobicity. Furthermore, the presence of ß-casein decreased the in vitro antioxidant capacities of the phenolic acids, especially for gallic acid. These findings provide some useful insights into the structure-activity relationships of the interaction between ß-casein and phenolic acids.

Analysis of inhibitory interaction between epigallocatechin gallate and alpha-glucosidase: A spectroscopy and molecular simulation study.

Alpha-glucosidase is one of the main enzymes responsible for digesting starch. Inhibiting its activity is therefore being targeted as a strategy for tackling diabetes. Certain food components have the potential to act as natural α-glucosidase (SCG) inhibitors, such as the polyphenols found in tea. In this study, epigallocatechin gallate (EGCG) was shown to strongly inhibit SCG activity (IC50 value = 3.7 × 10-5 M). Multi-spectroscopic binding molecular simulations indicated that EGCG spontaneously bound to SCG through a combination of hydrogen bonding and hydrophobic interactions. The hypothesis was supported by the results from intrinsic fluorescence quenching, conformational change, surface hydrophobicity decrease, and molecular docking analysis of the SCG after binding. Molecular docking provided powerful visual insights into the nature of the molecular interactions involved. This research provides important new information about the interaction mechanism of EGCG and SCG, which may be beneficial to the development of functional foods to prevent diabetes.

Amino acid-amidated pectin: Preparation and characterization.

There is interest in extending the functionality of natural polymers using simple chemical derivatization methods. In this study, the ability of pectin to react with various amino acids (40 °C) in aqueous solution without a catalyst was investigated, including glutamic acid, glycine, cysteine, lysine, and arginine. Only glycine, lysine, and arginine could form conjugates with pectin under these conditions. Amino acid-amidated pectin (AAAP) conjugates with a degree of amidation of 6.50% were prepared and characterized using elemental analysis, FT-IR, and 1H NMR. Size exclusion chromatography combined with multiangle light scattering and refractive index detection indicated pectin degradation occurred during the reaction. These results were supported by measurements of apparent viscosities. Scanning electron microscopy showed appreciable differences among the surface topographies of samples. Cell culture experiments showed the AAAP conjugates exhibited no cytotoxic effects. Our results suggested the AAAP conjugates may be suitable for use as new functional ingredients in food industry.