Release profile of amino acids encapsulated in solid lipid particles during in vitro oro-gastrointestinal digestion.

Some amino acids are known to mediate immune responses through gut microbiota metabolism in both humans and monogastric animals. However, through the diet, most free amino acids are absorbed in the small intestine and only a small quantity reaches the microbiota-rich colon. To enhance microbial metabolism of amino acids and their potential health benefits, encapsulation strategies are developed for their protection and delivery to the colon. So far, the main encapsulation systems for amino acids are based on solid lipid particles, but their fate within the digestive tract has never been fully clarified. In this study, we investigated the release of various amino acids (branched-chain amino acid mixture, or lysine, or tryptophan) loaded in solid lipid particles during in vitro oro-gastrointestinal digestion mimicking the piglet. The loaded solid lipid particles were fully characterized for their composition, thermal behavior, molecular structure, crystalline state, surface morphology, and particle size distribution. Moreover, we investigated the effect of particle size by sieving solid lipid particles into two non-overlapping size fractions. We found that amino acid release was high during the gastric phase of digestion, mainly controlled by physical parameters, namely particle size and crystalline state including surface morphology. Large particle size and/or smooth ordered particle indeed led to slower and lower release. Although lipid hydrolysis was significant during the intestinal phase of digestion, the impact of the crystalline state and surface morphology was also observed in the absence of enzymes, pointing to a dominant water/solute diffusion mechanism through these porous solid lipid particles.

Biochemical and physical-chemical characterisation of leaf proteins extracted from Cichorium endivia leaves.

This study provides a detailed characterisation of a leaf protein concentrate (LPC) extracted from Cichorium endivia leaves using a pilot scale process. This concentrate contains 74.1% protein and is mainly composed of Ribulose-1,5-BISphosphate Carboxylase/Oxygenase (RuBisCO). We show that the experimentally determined extinction coefficient (around 5.0 cm-1 g-1 L depending on the pH) and refractive index increment (between 0.27 and 0.39 mL g-1) are higher than the predicted ones (about 1.6 cm-1 g-1 L and 0.19 mL g-1, respectively). In addition, the UV-visible absorption spectra show a maximum at 258 nm. These data suggest the presence of non-protein UV-absorbing species. Chromatographic separation of the concentrate components in denaturing conditions suggests that RuBisCO SC may be covalently bounded to few phenolic compounds. Besides, the solubility of LPC proteins is higher than 90% above pH 6. Such high solubility could make LPC a good candidate as a functional food ingredient.

Structuring and functionalization of dispersions containing egg yolk, plasma and granules induced by mechanical treatments.

In this study, the impact of mechanical treatments on the physicochemical and emulsifying properties of hen egg yolk and its fractions plasma and granules has been assessed. Yolk, plasma, and granule dispersions at pH 4.0 and 0.75 M NaCl were subjected to rotor-stator and high-pressure pretreatments at different dynamic pressure levels: 30, 100, and 200 bar at 20 degrees C. Physicochemical characteristics (protein solubility, rheological behavior, and micro- and ultra-structures) and emulsifying properties (oil/water 60:40 emulsions: droplet size and flocculation, protein adsorption) of control dispersions and dispersions subjected to mechanical pretreatments (rotor-stator or high pressure) were compared. Homogenization at high pressures (100 and 200 bar) led to a decreased protein solubility and to an increase in apparent viscosity of yolk and plasma dispersions. These pressures certainly disrupted low-density lipoproteins (LDL) particles and generated aggregates of proteins liberated from LDL and livetins in the plasma fraction, and led to a moderated reorganization of the microstructure of granules. Despite the modifications observed in the pretreated plasma and granules dispersions, the oil droplet diameter and the bridging flocculation obtained in emulsions made with these dispersions were similar to that obtained with untreated dispersions. Results concerning interfacial protein adsorption suggested that preformed or natural aggregates at least partially persist at the oil-water interface.

Structures and rheological properties of hen egg yolk low density lipoprotein layers spread at the air-water interface at pH 3 and 7.

Low density lipoproteins (LDL) from egg yolk have a classical structure of lipoprotein with a core of neutral lipids surrounded by a monolayer of apoproteins and phospholipids. This structure collapses during adsorption and all constituents spread at the interface. To understand better the nature of the interactions between apoproteins and lipids at the interface, we have deposited LDL at an air-water interface and analysed the isotherms during their compression on a Langmuir trough. Then, these LDL films were studied by atomic force microscopy (AFM) imaging. To identify the protein and lipid structures, we imaged films before and after lipid solubilisation by butanol. To study the interactions in the LDL films, we have varied the pH, ionic strength and used simplified model systems. We also studied the correlation between observed structures and interfacial rheology of the film. The isotherms of interfacial LDL films were similar for pH 3 and 7, but their structures observed in AFM were different. At surface pressures below the transition corresponding to the demixion of apoprotein-neutral lipid complexes, the LDL film structure was not governed by electrostatic interactions. However, above this surface pressure transition (45mN/m), there was an effect of charge on this structure. Around the transition zone, the rheological properties of LDL films at pH 3 were different as a function of pH (viscous at pH 3 and visco-elastic at pH 7). So, the rheological properties of LDL films could be linked to the structures formed by apoproteins and observed in AFM.

Structure modification in hen egg yolk low density lipoproteins layers between 30 and 45 mN/m observed by AFM.

We have studied the structure of films made by low density lipoproteins (LDL) from hen egg yolk, which are composed of apoproteins, neutral lipids and phospholipids. These LDL have been deposited on air-water interface to form a monolayer which has been compressed to measure an isotherm using Langmuir balance. This isotherm presented three transitions (neutral lipid (surface pressure, pi=19 mN/m), apoprotein-lipid (pi=41 mN/m) and phospholipid (pi=51 mN/m) transitions). We have studied only the apoprotein-lipid transition. In order to observe the LDL film structure before (pi=30 mN/m) and after (pi=45 mN/m) the apoprotein-lipid transition, the formed films were transferred and visualised by atomic force microscopy (AFM). Our results have shown that the structures observed in the LDL film were different depending on the surface pressure. The apoproteins and neutral lipids appeared to be miscible up to the apoprotein-lipid transition, when demixing occurred. The structures observed after the apoprotein-lipid transition should be due to the demixing between apoproteins and neutral lipids. On the other hand, apoproteins and phospholipids seemed miscible whatever the surface pressure. Hence, the first transition (pi=19 mN/m) should be attributed to the free neutral lipid collapse; the second transition (pi=41 mN/m) should be attributed to the demixing of apoprotein-neutral lipid complexes; and the last transition (pi=51 mN/m) should be attributed to phospholipid collapse or to demixing of apoprotein-phospholipid complexes.

Hen egg yolk low-density lipoproteins film spreading at the air-water and oil-water interfaces.

Hen egg yolk is largely used as an ingredient in food emulsions due to its exceptional emulsifying properties. Low-density lipoproteins (LDL) are the main egg yolk constituents and the most important contributors to yolk emulsifying properties. To better understand the LDL adsorption mechanism and spreading at the interface, we extracted and studied LDL at different interfaces. At the air-water interface, the LDL film isotherm presents three transitions, and two were identified by each lipid class present in LDL. The last transition should be due to apoproteins-lipids complexes. During LDL adsorption, the presence of apoproteins at the LDL surface and the neutral lipid core is necessary. At pH 3 and pH 7, LDL are disrupted and spread quasi-similarly at the air-water interface, contrary to the oil-water interface where LDL spread more at pH 7 than at pH 3.

Liposomes as an alternative to egg yolk in stallion freezing extender.

Egg yolk is normally used as a protective agent to freeze semen of equine and other species. However, addition of egg yolk in extenders is not without disadvantages and the demand to find cryoprotective alternatives is strong. The objective of this study was to test the cryoprotective capacities of liposomes composed of egg yolk phospholipids. Two experiments were conducted: 1) the first to determine the optimal composition and concentration of liposomes to preserve post-thaw motility and membrane integrity of spermatozoa; 2) the second to assess in vivo the cryoprotective capacities of these liposomes. In Experiment 2, post-thaw motility and membrane integrity of spermatozoa were also analyzed. Experiment 1 demonstrated that liposomes composed of phospholipids E80 (commercial lecithins from egg yolk composed mainly of phosphatidylcholine and phosphatidylethanolamine) and of Hank's salts-glucose-lactose solution (E80-liposomes) were the most efficient in preserving post-thaw motility. The optimal concentration was 4 % (v/v). In Experiment 2, fertility rate after artificial insemination of semen frozen with E80-liposomes was 55 % (22/40) compared with 68 % (27/40) with the control extender containing egg yolk (EY) (p = 0.23). Post-thaw motility parameters were higher with EY than with E80-liposomes (p < 0.0001). For post-thaw membrane integrity no difference was observed between the two extenders (p = 0.08). Liposomes composed of egg yolk phospholipids appeared to be a promising alternative to replace egg yolk in semen freezing extenders in equine species.

Interfacial and oil/water emulsions characterization of potato protein isolates.

Interfacial and emulsifying properties of potato protein isolate (PPI) have been studied to evaluate its potential application to stabilize oil/water emulsions at two pH values (2 and 8). The amount, type, and solubility of proteins and the size of aggregates have been determined in aqueous dispersion. Air-water and oil-water interfacial properties (adsorption, spreading, and viscoelastic properties) have been determined as a function of concentration and pH using soluble phases of PPI. The behavior of PPI stabilized oil/water emulsions has been then analyzed by droplet size distribution measurements and interfacial concentration. PPI exhibits low solubility over a wide range of pH values, with the presence of submicrometer aggregates. The pH value exerts a negligible effect on interfacial tension (oil-water) or surface pressure (air-water) but displays very important differences in viscoelastic properties of the interfacial films formed between oil and water. In this sense, pH 8 provides a major elastic response at oil-water interfaces as compared to pH 2. In relation with this result, a much higher ability to produce fine and stable emulsions is noticed at pH 8 as compared to pH 2. Consequently, there is an evident relationship between the rheological properties of the oil-water interfacial films and the macroscopic emulsion behavior.