Effect of different oligomerization assemblies of γ-conglutin on its interaction behavior with vitexin.

BACKGROUND: Several different factors underlie the molecular mechanisms of phenolic compound-protein interactions. They include the environmental conditions. In the case of γ-conglutin, pH conditions translate directly into the adoption of two distinct oligomeric assemblies, i.e. hexameric (pH 7.5) or monomeric (pH 4.5). This paper reports research on the pH-dependent oligomerization of γ-conglutin in terms of its ability to form complexes with a model flavonoid (vitexin). RESULTS: Fluorescence-quenching thermodynamic measurements indicate that hydrogen bonds, electrostatic forces, and van der Waals interactions are the main driving forces involved in the complex formation. The interaction turned out to be a spontaneous and exothermic process. Assessment of structural composition (secondary structure changes and arrangement/dynamics of aromatic amino acids), molecular size, and the thermal stability of the different oligomeric forms showed that γ-conglutin in a monomeric state was less affected by vitexin during the interaction. CONCLUSION: The data show precisely how environmental conditions might influence phenolic compound-protein complex formation directly. This knowledge is essential for the preparation of food products containing γ-conglutin. The results can contribute to a better understanding of the detailed fate of this unique health-promoting lupin seed protein after its intake. © 2023 Society of Chemical Industry.

Effects of Heat Treatment Duration on the Electrical Properties, Texture and Color of Polymerized Whey Protein.

In this research effects of heat treatment duration on the electrical properties (zeta potential and conductivity), texture and color of polymerized whey protein (PWP) were analyzed. Whey protein solutions were heated for 30 min to obtain single-heated polymerized whey protein (SPWP). After cooling to room temperature, the process was repeated to obtain double-heated polymerized whey protein (DPWP). The largest agglomeration was demonstrated after 10 min of single-heating (zeta potential recorded as -13.3 mV). Single-heating decreased conductivity by 68% and the next heating cycle by 54%. As the heating time increased, there was a significant increase in the firmness of the heated solutions. Zeta potential of the polymerized whey protein correlated with firmness, consistency, and index of viscosity, the latter of which was higher when the zeta potential (r = 0.544) and particle size (r = 0.567) increased. However, there was no correlation between zeta potential and color. This research has implications for future use of PWP in the dairy industry to improve the syneretic, textural, and sensory properties of dairy products.

Polymer-Solvent Interactions in Modified Starches Pastes-Electrokinetic, Dynamic Light Scattering, Rheological and Low Field Nuclear Magnetic Resonance Approach.

Starch paste is a very complex dispersion that cannot be clearly classified as a solution, colloid or suspension and many factors affects its properties. As these ambiguities constitute a barrier to technological development, the aim of this study was to investigate the interaction of starch macromolecules with water by analysing the results of rheological properties, low field nuclear magnetic resonance (LF NMR), dynamic light scattering (DLS) and ζ potential analyses. Starch pastes with a concentration of 1%, prepared with distilled water and buffered to pH values of 2.5, 7.0 and 9.5 were analysed. It was proved that the pH buffering substantially decreased the values of consistency index but the pH value itself was not significant. LF NMR studies indicated that the dissolution of starch in water resulted in a reduction in spin-lattice as well as spin-spin relaxation times. Moreover, changes in relaxation times followed the patterns observed in rheological studies. Electrokinetic and DLS analyses showed that potential values are primarily influenced by the properties of the starches themselves and, to a lesser extent, by the environmental conditions. The conducted research also showed complementarity and, to some extent, substitutability of the applied research methods as well as exclusion chromatography (a method not used in this work).

Water content, critical micelle concentration of phospholipids and formation of association colloids as factors influencing autoxidation of rapeseed oil.

BACKGROUND: The exact mechanism of lipid autoxidation in vegetable oils, taking into account physical aspects of this phenomenon, including the role of association colloids, is still not fully understood. The purpose of this study was to consider changes in moisture content and DOPC phospholipid (1,2-dioleoyl-sn-glycero-3-phosphocholine) critical micelle concentration (CMC) in rapeseed oil during autoxidation as well as to find the relationship between these parameters and the accumulation of primary and secondary lipid oxidation products. RESULTS: The experiments were performed at initial oil humidity 220 ppm and 700 ppm, with DOPC below and above CMC. The increase in water concentration was favored by the presence of phospholipids above CMC and, at the same time, high initial water level, which favored oxidation processes and the creation of amphiphilic autoxidation products. At relatively high water level and low amphiphilic DOPC concentration, the growth of water content does not affect the concentration of oxidation products. CONCLUSION: Amphiphilic substances play a significant role in increasing the water content of oil. Autoxidation products may reduce CMC of DOPC, but water is able to compensate for the CMC-reducing effect of oxidation products. The presence of association colloids and initial water content play a crucial role in the oxidation process of rapeseed oil. The increase in water concentration does not cause a sufficiently large increase in the number of micelles or sufficiently significant changes in their structure to effect an increase in the level of oxidation products. The formation of micelles requires an appropriate content of both water and amphiphilic substances derived from seeds (phospholipids). © 2021 Society of Chemical Industry.

Heat-induced changes in lupin seed γ-conglutin structure promote its interaction with model phospholipid membranes.

A number of scientific data indicate that γ-conglutin can be internalised by different human cells and undergoes secretion from the seed in response to high temperature. In both of these cases, the protein must interact in some manner with biological membranes, however, the mechanisms underlying this phenomenon remain unknown. Herein, we found that the remarkable change of total surface hydrophobicity after appropriate heat treatment of γ-conglutin monomer led to its interaction with model membranes (liposomes). Before the interaction, the protein undergoes an intriguing thermal unfolding pattern which was studied based on a spectroscopic approach. Insight into the interaction mechanism with liposomes was possible thanks to applying two molecular probes that were differentially localised in the lipid bilayer. The results show that the thermal rearranged γ-coglutin monomer affects hydrocarbon chains in model membranes leading to their morphology change and disruption. The main driving force of this phenomenon is based on hydrophobic interaction.

Nanocellulose Production Using Ionic Liquids with Enzymatic Pretreatment.

Nanocellulose has gained increasing attention during the past decade, which is related to its unique properties and wide application. In this paper, nanocellulose samples were produced via hydrolysis with ionic liquids (1-ethyl-3-methylimidazole acetate (EmimOAc) and 1-allyl-3-methylimidazolium chloride (AmimCl)) from microcrystalline celluloses (Avicel and Whatman) subjected to enzymatic pretreatment. The obtained material was characterized using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), dynamic light scattering (DLS), scanning electron microscopy (SEM), and thermogravimetric analysis (TG). The results showed that the nanocellulose had a regular and spherical structure with diameters of 30-40 nm and exhibited lower crystallinity and thermal stability than the material obtained after hydrolysis with Trichoderma reesei enzymes. However, the enzyme-pretreated Avicel had a particle size of about 200 nm and a cellulose II structure. A two-step process involving enzyme pretreatment and hydrolysis with ionic liquids resulted in the production of nanocellulose. Moreover, the particle size of nanocellulose and its structure depend on the ionic liquid used.

Nutlets of Tilia cordata Mill. and Tilia platyphyllos Scop. - Source of bioactive compounds.

The aim of this study was to analyse contents of phytosterols, carotenoids, tocochromanols and fatty acid composition in oil from nutlets of linden. Standard determination methods for these compounds were applied using liquid and gas chromatography. In the analysed oils from small and large-leaved lime the dominant acids included linoleic (53.1 and 54.1%), oleic (18.5 and 22.2%), palmitic (15.3 and 11.8%). Linden seed oil was characterised by very high contents of phytosterols (2-2.5 g/100 g). ß-Sitosterol was the dominant phytosterol (80-83%). Additionally squalene was detected at 806 and 607 mg/100 g, respectively, for Tilia cordata and T.platyphyllos. It was found that linden seed oil contains tocopherols (93%) and tocotrienols (7%). In terms of tocochromanol contents this oil is surpassed only by wheat germ oil. In terms of the α-T equivalent these oils contain 95 (Tilia cordata) and 50.6 mg/100 g vitamin E (Tilia platyphyllos).

Evaluation of bioactive compounds in cereals. Study of wheat, barley, oat and selected grain products.

BACKGROUND: One of the requirements for proper nutrition and maintenance of good health is to supply the body, through diet, with an appropriately increased quantity of bioactive compounds. With this in mind, modern milling and baking industries keep introducing new types of products. The use of such additives as wholegrain flours and bran in baked products provided the basis for research in this paper. METHODS: The aim of the study was to conduct a qualitative and quantitative evaluation of marketable wheat, barley and oat grain, used as raw materials to produce dehulled kernels, ground grain, wholemeal flour and wheat flour type 550 (all-purpose or plain flour), as well as wheat bran. Additionally, analyses were performed to determine the chemical composition and contents of nutrients, selected bioactive compounds and antioxidant activity. RESULTS: The studied raw materials in commercial cereal differ in their chemical composition. Dehulling of wheat, barley and oat grains significantly contributed to the reduction of minerals, protein and total dietary fiber (TDF) contents, except for the amount of protein in dehulled wheat and oat grains. Oat bran, in contrast to other oat products, was characterized by the highest contents of minerals, protein, TDF, and the smallest amounts of saccharides and total starch. The lowest content of minerals was recorded in wheat flour type 550. Thermal processes affect the concentration of tocochromanols in the grain, with tocotrienols being more resistant to hydrothermal treatment than tocopherols. Grain dehulling also significantly decreased the total amount of tocochromanols in relation to the original grains. CONCLUSIONS: Wheat products differ in the contents of their individual components. They are characterized by high contents of tocochromanols, phenolic compounds and water-extractable arabinoxylans, with the exception of white refined wheat flour, which is mainly a source of saccharides. In the case of barley and oat products, the analysis showed no differences between these product groups. It was also shown that dehulling of barley and oat grain causes statistically significant differences in the contents of nutrients and natural antioxidants.

Preparation of Nanocellulose Using Ionic Liquids: 1-Propyl-3-Methylimidazolium Chloride and 1-Ethyl-3-Methylimidazolium Chloride.

Cellulose nanocrystals were prepared using ionic liquids (ILs), 1-ethyl-3-methylimidazolium chloride [EMIM][Cl] and 1-propyl-3-methylimidazolium chloride [PMIM][Cl], from microcrystalline cellulose. The resultant samples were characterized by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), dynamic light scattering (DLS), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The XRD results showed that nanocellulose obtained by treatment with both ILs preserved basic cellulose I structure, but crystallinity index of samples (except for Sigmacell treated with [EMIM][Cl]) was lower in comparison to the starting microcrystalline cellulose. The DLS results indicated noticeably smaller particle sizes of prepared cellulose for material treated with [PMIM][Cl] compared to cellulose samples hydrolyzed with [EMIM][Cl], which were prone to agglomeration. The obtained nanocellulose had a rod-like structure that was confirmed by electron microscopy analyses. Moreover, the results described in this paper indicate that cation type of ILs influences particle size and morphology of cellulose after treatment with ionic liquids.

Seed-Roasting Process Affects Oxidative Stability of Cold-Pressed Oils.

The oxidative stability of vegetable oils mainly depends on their fatty acid composition, their degree of unsaturation, and the presence of compounds with antioxidant activity. This paper reports on the effects of the process of roasting oil seeds, prior to pressing them, on the basic characteristics of the oils produced and their oxidative stability. The differential scanning calorimetry (DSC) technique was used to study the process of oxidation of the oil samples in an oxygen-flow cell. Chromatographic analysis revealed that roasting the seeds increased the levels of chlorophyll and ß-carotene in all the cold-pressed oils. Similar results were observed for the oil's antioxidant activity, measured by the scavenging 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical method. Our results also indicated that roasting seeds prior to pressing them for oil had a positive effect on the oil's stability, as determined by the DSC method. This manifested in both the extension of oxidation induction time and the final oxidation time.

The quality of cold-pressed rapeseed oil obtained from seeds of Brassica napus L. with increased moisture content.

BACKGROUND: The basic parameter influencing the quality of cold-pressed oil is the quality of seeds used  for pressing. Adverse moisture content and storage temperature of rape seeds may affect the quality of oil obtained from them. This paper presents the effects of increased rapeseed moisture content on the quality of the oil pressed. METHODS: The material used for the tests was rapeseed (canola) cv. PR 46 W14. The moisture content of the seeds was adjusted to 10%, 12% and 20%, and the seeds were stored at room temperature for 14 days. The samples were then dried to a seed moisture equal to 7% and oil was pressed from them. Acid and peroxide values, as well as the content of water, tocopherols and phenolic acids, were determined. In addition, a sensory analysis of the oil samples was carried out, and structural changes in the association colloids in the oil were determined using a fluorescent probe. RESULTS: With the increase in seed moisture, the increase in peroxide and acid values of the analyzed oils was recorded. A slight decrease in tocopherol content and a significant increase in phenolic acid concentration, depending on the seed moisture content, was observed. Sensory analysis showed odor sensory profile changes that probably indicate microflora development. CONCLUSIONS: The rapeseed moisture content has a crucial influence on the quality of oil obtained from them. Along with an increase in seed moisture, the possibility of developing undesirable microflora grows, which results in a deterioration in the quality of the obtained oil.

Physicochemical characteristics of the cold-pressed oil obtained from seeds of Fagus sylvatica L.

A physicochemical characteristic of the cold-pressed oil obtained from seeds of common beech (Fagus sylvatica L.) has been presented. This plant may be considered as unconventional oilseeds crops because of relatively high content of fat (27.25%). The analyzed beech seeds oil has been classified as oleic-linoleic acids oil with more than 76% percentage share of those species. Beech seeds oil contains 4.2% of gamma-linolenic acid (GLA). Unique characteristic is the high content of γ-tocopherol (75.4mg/100g) and δ-tocopherol (34.05mg/100g). γ-Tocopherol is effective scavengers of reactive nitrogen species and prevents DNA bases nitration, what makes beech seeds oil interesting raw material in the production of cosmetics. Additionally the content of carotenoids, very effective photooxidation inhibitors, is at high level in comparison with other cold-pressed oils. It was demonstrated that PCA analysis may help to determine the authenticity of oil obtained from beech seeds.

Fluorescence quenching studies on the interaction of catechin-quinone with CdTe quantum dots. Mechanism elucidation and feasibility studies.

Changes of the photoluminescent properties of QD in the presence of oxidized catechin (CQ) were investigated by absorption, steady-state fluorescence, fluorescence lifetime and dynamic light scattering measurements. Photoluminescence intensity and fluorescence lifetime was decreasing with increasing CQ concentration. Dynamic light scattering technique found the hydrodynamic diameter of QD suspension in water is in range of 45 nm, whereas in presence of CQ increased to mean values of 67 nm. Calculated from absorption peak position of excition band indicated on average QD size of 3.2 nm. Emission spectroscopy and time-resolved emission studies confirmed preservation of electronic band structure in QD-CQ aggregates. On basis of the presented results, the elucidated mechanism of QD fluorescence quenching is a result of the interaction between QD and CQ due to electron transfer and electrostatic attraction. The results of fluorescence quenching of water-soluble CdTe quantum dot (QD) capped with thiocarboxylic acid were used to implement a simple and fast method to determine the presence of native antioxidant quinones in aqueous solutions. Feasibility studies on this method carried out with oxidized catechin showed a linear relation between the QD emission and quencher concentration, in range from 1 up to 200 µM. The wide linear range of concentration dependence makes it possible to apply this method for the fast and sensitive detection of quinones in solutions.

Characterisation of different digestion susceptibility of lupin seed globulins.

This study describes in vitro digestion of lupin seed globulins by pancreatin, trypsin and chymotrypsin. Lupin seed globulins turned out to be almost totally susceptible to chymotrypsin digestion. When panceratin or trypsin were used for digestion of lupin seed globulins, γ-conglutin appeared to be resistant to proteolysis. Different fluorescence spectroscopic methods such as fluorescence anisotropy, fluorescence lifetimes and fluorescence quenching measurements were used for detailed characterisation of this phenomenon. A potential reason for γ-conglutin insensitivity to digestion may be related to the fact that lysine, as well as arginine, are positively charged at cell physiological pH. Simultaneously, flavonoids at this pH are partially ionised, which may lead to the occurrence of ionic interactions between these molecules at pH 7.5. The confirmation of this explanation may be the fact that γ-conglutin and vitexin form a static complex, which was observed using fluorescence quenching measurements.

The Interactions Between Rapeseed Lipoxygenase and Native Polyphenolic Compounds in a Model System.

The focus of the present research was to study inhibition of lipoxygenase activity by rapeseed native polyphenols and the interactions between those compounds and the enzyme. The enzyme and polyphenolic compounds (polyphenols, phenolic acids) were extracted from rapeseed (Brassica napus) varieties Aviso and PR45DO3. The total phenolic compounds concentration in tested rapeseed was 1,485-1,691 mg/100 g d.m. (dry matter) and the free phenolic acids content in both rapeseed varieties was about 76 µg/100 g d.m. The isolated proteins showed lipoxygenase activity. Prooxidant properties of phenolic compounds in the presence of lipoxygenase and linoleic acid were observed rather in the case of extracts containing a relatively high concentration of miscellaneous polyphenols. Antioxidant properties were recorded in the case of phenolic acid extracts which contain only 1.4-1.9% of phenolics present in raw phenolic extracts. We propose that the prooxidant effect of phenolic compounds comes from quinone and oxidized polyphenols formation. The observed antioxidant activity of phenolic acid extracts is probably due to their ability to scavenge free radicals formed from linoleic acid. However, reduction of lipoxygenase ferric to ferrous ions, which prevent the activation of the enzyme and inhibited its activity, was also observed.

Release of flavonoids from lupin globulin proteins during digestion in a model system.

Lupin seed globulin proteins form complexes with flavonoids, predominantly apigenin C-glycosides. Enzymes typical for the gastrointestinal tract were used to hydrolyze lupin seed globulins. Release of native flavonoids as a result of the proteolysis reaction was observed. Different analytical methods such as size exclusion chromatography, HPLC-MS, and fluorescence spectroscopy (steady-state fluorescence, fluorescence anisotropy, fluorescence lifetimes) were used for a detailed characterization of this phenomenon. Flavonoids liberated from lupin globulin proteins as a result of pancreatin-catalyzed digestion were bound by γ-conglutin resistant to this enzyme. Two possible mechanisms of this interaction may be suggested: hydrogen bonding between oligosaccharide chains of glycoproteins and the sugar moieties of the flavonoid glycosides or electrostatic attraction between positively charged γ-conglutin and flavonoids partially ionized at pH 7.5.

Spectroscopic studies of D-alpha-tocopherol concentration-induced transformation in egg phosphatidylcholine vesicles.

The effects of embedding up to 60 mol% of alpha-tocopherol (alpha-Toc) on the morphology and structure of the egg phosphatidylcholine (PC) membrane were studied using spectroscopic techniques. The resulting vesicles were subjected to turbidometric and dynamic light scattering measurements to evaluate their size distribution. The alpha-Toc intrinsic fluorescence and its quenching was used to estimate the tocopherol position in the membrane. Optical microscopy was used to visualize morphological changes in the vesicles during the inclusion of tocopherol into the 2 mg/ml PC membrane. The incorporation of up to 15 mol% of tocopherol molecules into PC vesicles is accompanied by a linear increase in the fluorescence intensity and the simultaneous formation of larger, multilamellar vesicles. Increasing the tocopherol concentration above 20 mol% induced structural and morphological changes leading to the disappearance of micrometer-sized vesicles and the formation of small unilamellar vesicles of size ranging from 30 to 120 nm, mixed micelles and non-lamellar structures.