Krzysztof Dwiecki

Spectroscopic studies of D-α-tocopherol concentration-induced transformation in egg phosphatidylcholne vesicles

The effects of embedding up to 60 mol% of α-tocopherol (α-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 α-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.

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.

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.

A review of methods used for investigation of protein–phenolic compound interactions

Interactions between the different compounds present in foods are common and have influence on the nutritional and functional properties of food products. Among a wide range of these interactions, the formation of complexes between proteins and phenolic compounds seems to be the most important issue. Complexation of the phenolic compounds with proteins can be analysed considering several aspects. These complexes might strongly affect nutritional potential of polyphenols by masking their antioxidant capacity, and on the other hand might have influence on the structure of proteins which may cause their precipitation or decrease susceptibility to digestion. The complexity of protein–phenolic compound interactions is a challenge for food analysts and forced researchers to establish a wide range of analytical methods, allowing determination of complexes formation. The main aim of this review is to give researchers an overview of the currently used methods that can be applied to study the interactions between proteins and phenolic compounds.

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.

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.

Modification of soybean and lupine sprouting conditions: influence on yield, ROS generation, and antioxidative systems

Formation of free radicals’ and antioxidants’ biosynthesis during seeds sprouting strongly depends on plant growth conditions, especially while sprouts are fortified in iron. The influence of watering with FeSO4 solution on soybean and lupine sprouting, yield of iron accumulated in plant tissue, as well as free radical (reactive oxygen and nitrogen species) synthesis and antioxidant activity (capacity) were the objective of this study. Optimal iron accumulation and biomass efficiency were obtained for 3 day watering with water, followed by abiotic stress application (Fe2+ ions). For lupine sprouts, maximal radical formation (up to ~ 5.7 a.u.) took place on the fifth day of culturing in these conditions, while, for soybean sprouts, continuous increase in their activity was observed until the seventh day (up to ~ 5.8 a.u.). Total antioxidant activity in lupine sprouts increased definitely, even more than three times during 7 days of lupine growing, but, for soybean, it was almost steady. The same trend was observed in phenolic compounds and flavonoids contents. For soybean sprouts, these antioxidants were almost steady, while, for lupine sprouts in these conditions, the intensive synthesis of these antioxidants was observed. In response to high iron content applied from the fourth day in the cultivation media, lupine sprouts synthesize low molecular antioxidants effectively. For soybean sprouts, another mechanism of defense against oxidative stress should be expected.