Advances in Understanding the Antioxidant and Antigenic Properties of Egg-Derived Peptides.

Pepsin, trypsin and proteinase K were used in the present study to hydrolyse the proteins from whole eggs, yolks or whites, and the resulting hydrolysates were characterised in terms of antioxidant and IgE-binding properties, using a combination of in vitro and in silico methods. Based on the degree of hydrolysis (DH) results, the egg yolk proteins are better substrates for all the tested enzymes (DH of 6.2-20.1%) compared to those from egg whites (DH of 2.0-4.4%). The SDS-PAGE analysis indicated that pepsin and proteinase K were more efficient compared to trypsin in breaking the intramolecular peptide bonds of the high molecular weight egg proteins. For all the tested substrates, enzyme-assisted hydrolysis resulted in a significant increase in antioxidant activity, suggesting that many bioactive peptides are encrypted in inactive forms in the parent proteins. The hydrolysates obtained with proteinase K exhibited the highest DPPH radical scavenging activity (124-311 µM Trolox/g protein) and the lowest residual IgE-binding capacity. The bioinformatics tools revealed that proteinase K is able to break the integrity of the main linear IgE-binding epitopes from ovalbumin and ovomucoid. It can be concluded that proteinase K is a promising tool for modulating the intrinsic properties of egg proteins.

Alternative Processing Options for Improving the Proteins Functionality by Maillard Conjugation.

Conjugation of the proteins with carbohydrates, occurring in the early stages of the Maillard reactions, received increased attention because of the high potential to ensure the improvement of the biological activity and functional properties of the proteins of different origins. The Maillard conjugates are conventionally formed through wet or dry heating, but the use of alternative technologies involving ultrasound, microwave, pulsed electric fields, high-pressure, or electrodynamic treatments appears to be efficient in accelerating the reaction steps and limiting the formation of toxic compounds. An overview of the mechanisms of these processing technologies, the main parameters influencing the Maillard conjugate formation, as well as their advantages and disadvantages, is provided in this paper. Different strategies employing these alternative technologies are reported in the literature: as pretreatment of the proteins, either alone or in admixture with the carbohydrates, followed by conventional heating, as a single alternative treatment step, or as a combination of heating and alternative processing. The desired functional properties of the proteins can be achieved by selecting the appropriate processing strategy and optimizing the reaction parameters. Moreover, alternative technologies can be exploited to obtain Maillard conjugates with remarkable biological activity in terms of antioxidant, antimicrobial, antihypertensive, anti-inflammatory, antimutagenic, or bifidogenic properties.

Advanced interactional characterization of the inhibitory effect of anthocyanin extract from Hibiscus sabdariffa L. on apple polyphenol oxidase.

In this study, a comprehensive approach to advance the inhibitory effect of Hibiscus sabdariffa extract on apple polyphenol oxidase (PPO) was performed. PPO was extracted, purified, and characterized for optimal activity, whereas response surface methodology generated a quadratic polynomial model to fit the experimental results for hibiscus extraction. The optimum conditions allowed to predict a maximum recovery of anthocyanins (256.11 mg delphinidin-3-O-glucoside/g), with a validated value of 272.87 mg delphinidin-3-O-glucoside/g dry weight (DW). The chromatographic methods highlighted the presence of gallic acid (36,812.90 µg/g DW extract), myricetin (141,933.84 µg/g DW extract), caffeic acid (101,394.07 µg/g DW extract), sinapic acid (1157.46 µg/g DW extract), kaempferol (2136.76 µg/g DW extract), and delphinidin 3-O-ß-d-glucoside (226,367.08 µg/g DW extract). The inactivation of PPO followed a first-order kinetic model. A temperature-mediated flexible fit between PPO and anthocyanins was suggested, whereas the molecular docking tests indicated that PPO is a good receptor for cafestol, gallic acid, and catechin, involving hydrophobic and hydrogen bond interactions. PRACTICAL APPLICATION: It is well known that enzymatic browning is one of the most important challenges in the industrial minimal processing of selected fruit and vegetable products. Novel inhibitors for polyphenol oxidase are proposed in this study by using an anthocyanin-enriched extract from Hibiscus sabdariffa L. Based on our results, combining the chemical effect of phytochemicals from hibiscus extract with different functional groups with minimal heating could be an interesting approach for the development of a new strategy to inhibit apple polyphenol oxidase.

Proteases as Tools for Modulating the Antioxidant Activity and Functionality of the Spent Brewer's Yeast Proteins.

The functionality of the peptides obtained through enzymatic hydrolysis of spent brewer's yeast was investigated. Hydrolysis was carried out for 4-67 h with bromelain, neutrase and trypsin. The resulting hydrolysates were characterized in terms of physical-chemical, antioxidant and techno-functional properties. The solid residues and soluble protein contents increased with the hydrolysis time, the highest values being measured in samples hydrolyzed with neutrase. Regardless of the hydrolysis time, the maximum degree of hydrolysis was measured in the sample hydrolyzed with neutrase, while the lowest was in the sample hydrolyzed with trypsin. The protein hydrolysate obtained with neutrase exhibited the highest DPPH radical scavenging activity (116.9 ± 2.9 µM TE/g dw), followed by the sample hydrolyzed with trypsin (102.8 ± 2.7 µM TE/g dw). Upon ultrafiltration, the fraction of low molecular weight peptides (<3 kDa) released by bromelain presented the highest antioxidant activity (50.06 ± 0.39 µM TE/g dw). The enzymes influenced the foaming properties and the emulsions-forming ability of the hydrolysates. The trypsin ensured the obtaining of proteins hydrolysate with the highest foam overrun and stability. The emulsions based on hydrolysates obtained with neutrase exhibited the highest viscosity at a shear rate over 10 s-1. These results indicate that the investigated proteases are suitable for modulating the overall functionality of the yeast proteins.

CO2 supercritical extraction and microencapsulation of oleoresins from rosehip fruits for getting powders with multiple applications.

The supercritical fluids extraction (SFE) was used to extract the oleoresins from rosehip, followed by an in-depth phytochemical analysis and the development of two design-customized powders for different food and pharmaceutical applications. The SFE experiments allowed obtaining an oleoresins extraction yield of 11.85%. Two fractions were separated (S40 and S45), with significantly different phytochemical profile (p < 0.05), highlighting the efficiency of extraction of fatty acids in S40 extract, whereas the extraction of polyphenols, phytosterols, carotenoids and polyphenols was favored in S45 extract. The phytochemical profile revealed that the linoleic acid (C18:2) and α-linolenic acid (C18:3) represented approximatively 82% and 58% from the total fatty acid content in S40 and S45, respectively. α-Tocopherol and γ-tocopherol prevailed in both extract fractions, with a higher concentration in S45 (229.66 mg/g dry matter (DM) and 112.36 mg/g DM, respectively), whereas ß-sitosterol was the major phytosterol in S45 fraction (118.75 mg/g DM). The S40 fraction was used to design two microencapsulated powders, by combining emulsification, complex coarcevation and freeze-drying. In order to develop new wall materials, with unique properties, the soy protein isolates were used for cross-linked reactions, by using an approach in one step (transglutaminase mediated) (coded as N) and two-steps (heat-induced and transglutaminase mediated) (coded as T). The N powder showed a better phytochemical content, leading to a higher antioxidant activity (5.27 mM Trolox equivalents/g DM), whereas for variant T, the bioactive were apparently doubled encapsulated.

Value-added salad dressing enriched with red onion skin anthocyanins entrapped in different biopolymers.

In this study, red onion skin extract was used to obtain food ingredients. Complex biopolymeric matrices were dissolved in the anthocyanin-rich aqueous extract, followed by gelation and freeze-drying. Powders were characterized regarding encapsulation efficiency (EE), phytochemical content, color, antioxidant activity, and microstructure. Storage and simulated digestion stability were also assessed. Two powders with high contents of bioactives and antioxidant activity were obtained. The highest EE was acquired for the powder with a higher polysaccharides concentration (V2). In addition, V2 exhibited the best storage stability. The in vitro studies demonstrated that increased carbohydrate concentration delivers the best anthocyanins protection. To prove its functionality, V2 was added to a salad dressing. The addition of powder has improved the concentration of biologically active compounds and the antioxidant activity of the salad dressings. These results support the assumption that microencapsulation may deliver bioactives from red onion skin as functional ingredients for value-added foods.

Investigations on Functional and Thermo-Mechanical Properties of Gluten Free Cereal and Pseudocereal Flours.

Seven commercial gluten-free (rice, oat, sorghum, foxtail millet, amaranth, quinoa, and buckwheat) flours were investigated in this study from the point of view of thermo-mechanical properties and solvent retention capacity (SRC). Each flour was used to prepare doughs with specific water absorption (WA) to get a consistency of 1.1 Nm (WA1) and doughs with WA2 levels higher than 85% to ensure a sufficient amount of water in the system for allowing the hydration of all components of the flours. Different correlations were established between proteins, ash, pentosans, damaged starch, and amylose contents on the one hand, and the capacity of the flour samples to retain different solvents such as sucrose, sodium carbonate and CaCl2 on the other hand. Although no significant correlation was found between the protein content of the flours and lactic acid-SRC, the mechanical weakening of the protein was significantly correlated with lactic acid-SRC for both tested WA levels. The doughs with WA1 had higher starch gelatinization and hot gel stability values compared to the corresponding dough systems with a higher water amount. Moreover, lower starch retrogradation and setback torques were obtained in the case of the dough prepared with higher amounts of water.

Value-Added Crackers Enriched with Red Onion Skin Anthocyanins Entrapped in Different Combinations of Wall Materials.

The objective of this study was to encapsulate anthocyanins from red onion skins into different biopolymeric matrices as a way to develop powders with multifunctional activities. Two different variants of powders were obtained using a combination of gelation and freeze-drying techniques and characterized by encapsulation efficiency, antioxidant activity, phytochemical content, and color. Stability during storage and the bioavailability of anthocyanins in the in vitro simulated digestion were also examined. Powder V2, with a higher concentration of polysaccharides than V1, allowed a better encapsulation efficiency (90.53 ± 0.29%) and good stability during storage. Both variants had a high content of phytochemicals and antioxidant activity. In vitro investigations proved that an increased polysaccharides concentration offers the best protection for anthocyanins. Thus, a controlled release of the anthocyanins in the intestinal medium was achieved. The powder with the highest encapsulation efficiency was added to crackers, followed by phytochemical characterization to assess its potential added value. The addition of the micro-particles improved the functional characteristics such as antioxidant activity, showing its suitability for the development of bakery products. The attained results may bring implicit benefits to consumers, who can benefit from improved bioactive concentrations in foodstuffs, with significant health benefits.

Advanced Composites Based on Sea Buckthorn Carotenoids for Mayonnaise Enrichment.

This study aimed at the extraction and encapsulation of the carotenoids from sea buckthorn fruits and obtaining value-added mayonnaise. First, the carotenoids from sea buckthorn fruits were extracted using ultrasound-assisted extraction. Then, they were microencapsulated through complex coacervation and freeze-drying techniques using different wall material combinations. Two powders were obtained and analyzed in terms of encapsulation efficiency, total carotenoid content, antioxidant activity, stability of phytochemicals and color, morphological structure, and in vitro digestibility. All results pointed out that the carotenoid molecules were successfully encapsulated within the mixture of alginate, agar, and chitosan, with a 61.17 ± 0.89% encapsulation efficiency. To probe the functionality, the powder was added into mayonnaise in 2.5% and 5% amounts. The obtained mayonnaise samples were characterized in terms of phytochemical and antioxidant activity properties with their storage stability and texture, color, and sensory characteristics. A significant increase of total carotenoid content and antioxidant activity compared to the control sample was observed. The addition of powder also led to improved texture by increasing the firmness and adhesion. In addition, the sensory evaluation indicated an improved color and overall acceptability of the value-added mayonnaise. Thus, sea buckthorn extracts may be considered as valuable ingredients for the development of added-value food products.

Thermal Stability and Inhibitory Action of Red Grape Skin Phytochemicals against Enzymes Associated with Metabolic Syndrome.

The present study focuses on heat-induced structural changes and the degradation kinetics of phytochemicals and antioxidant activity of red grape skin extract. The thermal degradation of anthocyanins, flavonoids, polyphenols, and antioxidant activity followed a first-order kinetic model, increasing with temperature due to the intensification of the degradation process. The activation energy (Ea) highlighted this phenomenon. Likewise, the kinetic and thermodynamic parameters certified the irreversible degradation of the bioactive compounds from the skin of the Babeasca neagra grape variety. Both temperature and duration of heating had a significant impact on the content of bioactive compounds. In addition, the red grape skin extract inhibited certain enzymes such as α-amylase, α-glucosidase, lipase, and lipoxygenase, which are associated with metabolic syndrome and inflammation. Further knowledge on the possible inhibition mechanisms exerted by the major anthocyanins found in red grape skin extract on the metabolic syndrome-associated enzymes was gathered upon running molecular docking tests. Detailed analysis of the resulting molecular models revealed that malvidin 3-O-glucoside binds in the vicinity of the catalytic site of α-amylase and lipase, whereas no direct contact with catalytic amino acids was identified in the case of α-glucosidase and lipoxygenase.

Whey Proteins Isolate-Based Biopolymeric Combinations to Microencapsulate Supercritical Fluid Extracted Oleoresins from Sea Buckthorn Pomace.

In this study, high-value, carotenoid-rich oleoresin obtained by supercritical carbon dioxide (SFE-CO2) extraction was used to develop five variants of microencapsulated delivery system, based on whey proteins isolate (WPI), in combination with inulin (I), pectin (P) or lactose (L). The WPI:I and WPI:L variants were also obtained by conjugation via Maillard reaction. The microencapsulation of the SFE-CO2 sea buckthorn pomace oleoresin was performed by emulsion, complex coacervation and freeze-drying, which allowed for the obtaining of five powders, with different phytochemicals profile. The WPI:I conjugate showed the highest level of total carotenoids, whereas the counterpart WPI:L showed the highest content in linoleic acid (46 ± 1 mg/g) and palmitoleic acid (20.0 ± 0.5 mg/g). The ß-tocopherol and ß-sitosterol were identified in all variants, with the highest content in the conjugated WPI:L variant. Both WPI:L and WPI:I conjugate samples presented similar IC50 value for inhibitory activity against pancreatic lipase and α-amylase; the highest activity was observed for the conjugated WPI:I. The WPI:P combination allowed the highest release of carotenoids in the gastro-intestinal environment. All the powders exhibited poor flowing properties, whereas water activity (aw) ranged from 0.084 ± 0.03 to 0.241 ± 0.003, suggesting that all variants are stable during storage. In case of solubility, significant differences were noticed between non-heated and glycated samples, with the highest value for the WPI:I and the lowest for glycated WPI:I. The structural analysis revealed the presence of finer spherosomes in WPI:I and WPI:L, with a reduced clustering capacity, whereas the particles in the conjugated samples were more uniform and aggregated into a three-dimensional network.

Whey Protein Isolate-Xylose Maillard-Based Conjugates with Tailored Microencapsulation Capacity of Flavonoids from Yellow Onions Skins.

The objective of this study is to encapsulate flavonoids from yellow onion skins in whey protein isolates (WPI) and xylose (X), by Maillard-based conjugates, as an approach to improve the ability to entrap flavonoids and to develop powders with enhanced antioxidant activity. WPI (0.6%, w/v) was conjugated to X (0.3%, w/v) through the Maillard reaction at 90 °C for 120 min, in the presence of a flavonoid-enriched extract. Two variants of powders were obtained by freeze-drying. The glycation of WPI allowed a better encapsulation efficiency, up to 90.53 ± 0.29%, corresponding to a grafting degree of 30.38 ± 1.55%. The molecular modelling approach was used to assess the impact of X interactions with α-lactalbumin and ß-lactoglobulin on the ability of these proteins to bind the main flavonoids from the yellow onion skins. The results showed that X might compete with quercetin glucosides to bind with α-lactalbumin. No interference was found in the case of ß-lactoglobulin. The microstructural appearance of the powders revealed finer spherosomes in powder with WPI-X conjugates via the Maillard reaction. The powders were added to nachos, followed by a phytochemical characterization, in order to test their potential added value. An increase in antioxidant activity was observed, with no significant changes during storage.

Bioactive's Characterization, Biological Activities, and In Silico Studies of Red Onion (Allium cepa L.) Skin Extracts.

This study aimed to investigate the thermal stability and biological activities of the phytochemicals from the red onion skins extract, which are a rich source of anthocyanins. Eight anthocyanins were identified in the extract by high-performance liquid chromatography, the most abundant ones being cyanidin 3-O-laminaribioside and cyanidin 3-O-(6″-malonoyl-laminaribioside). The study also involved the assessment of the thermal degradation kinetics of anthocyanins and antioxidant activity in the 75-155 °C temperature range. The thermal degradation kinetics was described using the first-order kinetics model. In terms of thermal stability, increasing the temperature resulted in lower half-life values (t1/2) and higher degradation rate constant values (k) for both anthocyanins and antioxidant activity. The thermodynamic parameters revealed that the phytochemicals' degradation is a non-spontaneous and endothermic reaction. Furthermore, the inhibitory effect of the extract was investigated against the enzymes affiliated with metabolic syndrome, oxidative stress, and inflammatory process diseases. Thus, we also demonstrated that the red onion skins extract exerted inhibitory activity on α-glucosidase, α-amylase, lipase, and lipoxygenase. Considering the high content of bioactives and various biological properties, the red onion skins extract is suitable for multiple applications.

Preliminary Investigations into the Use of Amylases and Lactic Acid Bacteria to Obtain Fermented Vegetable Products.

Legumes are valuable sources of proteins and other functional components. However, the high starch content can be an impediment in developing new vegan food formulations. Enzyme-assisted hydrolysis was used to hydrolyze the starch from chickpea and broad bean vegetable milk to further develop vegetable lactic acid-fermented products. The antioxidant activity of legumes was tested, and it was observed that the overall antioxidant activity (DPPH radical scavenging ability) significantly increased after enzyme-assisted hydrolysis while total phenols content decreased. The obtained vegetable milk was then fermented using exopolysaccharides-producing lactic acid bacteria. A significant decolorization was observed after fermentation in the case of broad bean-based products. Rheological behavior of the fermented products was determined using small amplitude oscillatory measurements and the three-interval thixotropy test. Results showed higher complex viscosity values for broad bean-based products, which displayed a weak gel-like structure. The starter cultures used for vegetable milk samples fermentation influenced the resistance to flow.

Encapsulation of Anthocyanins from Cornelian Cherry Fruits Using Heated or Non-Heated Soy Proteins.

In the current study, the effect of temperature on the potential of soy proteins to ensure the encapsulation and gastric stability of bioactives, such as anthocyanins from cornelian cherry fruits, was investigated. The powders obtained after freeze-drying were analyzed in relation to flow properties, encapsulation retention and efficiency, stability in simulated gastrointestinal medium, color, and morphology. Preheating the soy proteins generated a powder with low density. Powders obtained with native soy proteins allowed the highest encapsulation efficiency and the lowest was obtained when using preheated soy proteins. The heat treatment of the mixture of soy proteins and cornelian cherry fruits prior to encapsulation generated powders with the highest lightness and the lowest intensity of red shades among all samples. The in vitro experiments revealed that the highest protection in simulated gastric environment was provided when protein was heat treated either alone or in combination with bioactives to be encapsulated. The morphological analysis highlighted that powders consisted of large and rigid structures.

Bovine ß-lactoglobulin peptides as novel carriers for flavonoids extracted with supercritical fluids from yellow onion skins.

Our study describes in detail the binding mechanism between the main flavonoids that were extracted from onion skins by supercritical CO2 and peptides from whey proteins, from the perspective of obtaining multifunctional ingredients, with health-promoting benefits. The supercritical CO2 extract had 202.31 ± 11.56 mg quercetin equivalents/g DW as the major flavonoid and antioxidant activity of 404.93±1.39 mM Trolox/g DW. The experiments on thermolysin-derived peptides fluorescence quenching by flavonoids extract allowed estimating the binding parameters, in terms of binding constants, and the number of binding sites. The thermodynamic analysis indicated that the main forces involved in complex formation were hydrogen bonds and van der Waals interactions. Molecular docking tests indicated that peptide fluorescence quenching upon gradual addition of onion skin extract might be due to flavonoids binding by Val15 -Ser21 . All 7 to 14 amino acids long peptides appeared to have affinity toward quercetin-3,4'-O-diglucoside and quercetin-4'-O-monoglucoside. The study is important as a potential solution for reuse of valuable resources, underutilized, such as whey peptides and yellow onion skins flavonoids for efficient microencapsulation, as a holistic approach to deliver healthy and nutritious food. PRACTICAL APPLICATION: A growing interest was noticed in the last years in investigating the interactions between proteins and different biologically active compounds, such as to provide knowledge for efficient development of new food, pharmaceutical, and cosmetic products. Recent studies suggest that flavonoid-protein complexes may be designed to improve the functional performance of the flavonoids. The results obtained in this study bring certain benefits in terms of exploiting the bioactive potential of both flavonoids and bioactive peptides, for developing of formulas with improved functional properties.

Thermal Degradation Kinetics of Anthocyanins Extracted from Purple Maize Flour Extract and the Effect of Heating on Selected Biological Functionality.

The thermal degradation of the anthocyanins and antioxidant activity in purple maize extracts was determined between 80 and 180 °C. The anthocyanins were found to be thermostable in the temperature range of 80 to 120 °C, whereas at higher temperatures the thermal degradation of both anthocyanins and antioxidant activity followed a first-order kinetic model. The z-values started from 61.72 ± 2.28 °C for anthocyanins and 75.75 ± 2.87 °C for antioxidant activity. The conformational space of pairs of model anthocyanin molecules at 25 and 180 °C was explored through a molecular dynamics test, and results indicated the occurrence of intermolecular self-association reactions and intramolecular co-pigmentation events, which might help explaining the findings of the degradation kinetics. The relationship between thermal degradation of anthocyanins and antioxidant activity and the in vitro release was further studied. The unheated extracts showed a high stability under gastric environment, whereas after heating at 180 °C, the digestion ended quickly after 60 min. After simulated intestinal digestion, the anthocyanins were slowly decreased to a maximum of 12% for the unheated extracts, whereas an 83% decrease was found after preliminary heating at 180 °C. The thermal degradation of anthocyanins was positively correlated with the in vitro decrease of antioxidant activity.

Fostering Lavender as a Source for Valuable Bioactives for Food and Pharmaceutical Applications through Extraction and Microencapsulation.

Lavender flowers were used in this study as a source of phytochemicals as naturally occurring antioxidants. Two different extraction techniques were applied, such as ultrasound-assisted (UAE) and supercritical fluids (SCE) methods. The comparative evaluation of the phytochemicals profile evidenced a higher content of chlorophyll a and b of 5.22 ± 0.12 mg/g dry weight (D.W.) and 2.95 ± 0.16 mg/g D.W, whereas the carotenoids content was 18.24 ± 0.04 mg/g D.W. in the SCE extract. Seven main compounds were found in both extracts: ß-linalool, eucalyptol, linalool acetate, ß-trans-ocimene, and limonene in SCE and linalool acetate, ß-linalool, 6-methyl-2-(2-oxiranyl)-5-hepten-2-ol, linalool oxide, lavandulyl acetate and camphor in UAE. The (n-3) acids had a higher contribution in SCE. The extracts were microencapsulated in different combinations of wall materials based on polysaccharides and milk proteins. The four variants showed different phytochemical and morphological profiles, with a better encapsulating efficiency for proteins (up to 98%), but with a higher content of encapsulated carotenoids for polysaccharides, the latter showing remarkable antimicrobial activity against selected microorganisms. Carboxymethyl cellulose and whey proteins led to a double encapsulation of lipophilic compounds. The powders were tested in two food matrices as ingredients, with multiple targeted functions, such as flavoring, antimicrobial, antioxidant activity that can successfully replace synthetic additives.

Tailoring the Health-Promoting Potential of Protein Hydrolysate Derived from Fish Wastes and Flavonoids from Yellow Onion Skins: From Binding Mechanisms to Microencapsulated Functional Ingredients.

This study focuses on combining different bioprocessing tools in order to develop an in-depth engineering approach for enhancing the biological properties of two valuable food by-products, namely fish waste and yellow onion skins, in a single new bioactive formulation. Bone tissue from phytophagous carp (Hypophthalmichthys molitrix) was used to obtain bioactive peptides through papain-assisted hydrolysis. The peptides with molecular weight lower than 3 kDa were characterized through MALDI-ToF/ToF mass spectrometry and bioinformatics tools. As a prerequisite for microencapsulation, the ability of these peptides to bind the flavonoids extracted from yellow onion skins was further tested through fluorescence quenching measurements. The results obtained demonstrate a considerable binding potency with a binding value of 106 and also the presence of one single or one class of binding site during the interaction process of flavonoids with peptides, in which the main forces involved are hydrogen bonds and van der Waals interactions. In the freeze-drying microencapsulation process, an efficiency for total flavonoids of 88.68 ± 2.37% was obtained, considering the total flavonoids and total polyphenols from the powder of 75.72 ± 2.58 quercetin equivalents/g dry weight (DW) and 97.32 ± 2.80 gallic acid equivalents/g DW, respectively. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test on the L929 cell line cultivated in the presence of different concentrations of microencapsulated samples (0.05-1.5 mg/mL) proved no sign of cytotoxicity, the cell viability being over 80% for all the samples.

The Interaction of Bovine ß-Lactoglobulin with Caffeic Acid: From Binding Mechanisms to Functional Complexes.

In this study, the interaction of native and transglutaminase (Tgase) cross-linked ß-lactoglobulin (ß-LG) with caffeic acid (CA) was examined, aiming to obtain functional composites. Knowledge on the binding affinity and interaction mechanism was provided by performing fluorescence spectroscopy measurements, after heating the native and cross-linked protein at temperatures ranging from 25 to 95 °C. Regardless of the protein aggregation state, a static quenching mechanism of intrinsic fluorescence of ß-LG by CA was established. The decrease of the Stern-Volmer constants with the temperature increase indicating the facile dissociation of the weakly bound complexes. The thermodynamic analysis suggested the existence of multiple contact types, such as Van der Waals' force and hydrogen bonds, between ß-LG and CA. Further molecular docking tests indicated the existence of various CA binding sites on the ß-LG surface heat-treated at different temperatures. Anyway, regardless of the simulated temperature, the CA-ß-LG assemblies appeared to be unstable. Compared to native protein, the CA-ß-LG and CA-ß-LGTgase complexes (ratio 1:1) exhibited significantly higher antioxidant activity and inhibitory effects on α-glucosidase, α-amylase, and pancreatic lipase, enzymes associated with metabolic syndrome. These findings might help the knowledge-based development of novel food ingredients with valuable biological properties.