Walnut (Juglans regia L.) kernel oil bodies recovered by aqueous extraction for utilization as ingredient in food emulsions: Exploration of their microstructure, composition and the effects of homogenization, pH, and salt ions on their physical stability.

Natural oil-in-water emulsions containing plant oil bodies (OBs), also called oleosomes, rich in health-promoting omega-3 polyunsaturated fatty acids (ω3 PUFA) are of increasing interest for food applications. In this study, we focused on walnut kernel OBs (WK-OBs) and explored their microstructure, composition and physical stability in ionic environments as well as the impact of homogenization. A green process involving aqueous extraction by grinding of WK allowed the co-extraction of OBs and proteins, and centrifugation was used to recover the WK-OBs. Confocal laser scanning microscopy images showed the spherical shape of WK-OBs with an oil core envelopped by a layer of phospholipids (0.16 % of lipids) and embedded proteins. Their mean diameter was 5.1 ± 0.3 µm. The WK-OBs contained 70.1 % PUFA with 57.8 % ω6 linoleic acid and 12.3 % ω3 α-linolenic acid representing 68 % and 11.6 % of the total fatty acids in the sn-2 position of the triacylglycerols (TAG), respectively. Trilinolein was the main TAG (23.1 %). The WK-OBs also contained sterols (1223 ± 33 mg/kg lipids; 86 % ß-sitosterol), carotenoids (0.62 ± 0.01 mg/kg lipids; 49.2 % ß-carotene), and tocopherols (322.7 ± 7.7 mg/kg lipids; 89 % γ-tocopherol), confirming their interest as health-promoting ingredients. The decrease in the size of WK-OBs under high-pressure homogenization avoided phase separation upon storage. The anionic WK-OB surface at neutral pH was affected by stressful ionic environments (pH, NaCl, CaCl2), that induced aggregation of WK-OBs and decreased the physical stability of the emulsions. Emulsions containing WK-OBs are promising to diversify the market of the ω3-rich plant-based food products and beverages.

Deciphering the properties of hemp seed oil bodies for food applications: Lipid composition, microstructure, surface properties and physical stability.

Hemp seed oil bodies (HSOBs) are of growing interest in response to the demand of consumers for healthy and natural plant-based food formulations. In this study, we used minimal processing including aqueous extraction by grinding and centrifugation to obtain HSOBs. We determined the lipid composition of HSBOs, their microstructure, and the impact of the homogenization pressure, pH and minerals on their surface properties and the physical stability of the emulsions. HSOBs contain high levels of well-balanced PUFA with LA/ALA = 2.9, γ-tocopherol, lutein and phytosterols. The mean diameter of HSOBs was 2.3 ± 0.1 µm with an isoelectric point in the range of pH 4.4 to 4.6. Homogenization of hemp seed extracts induced a decrease in the size of HSOBs but did not eliminate the sedimentation of the protein bodies composed of the globulin edestin. By changing the surface properties of HSOBs, pH values below 6 and NaCl induced the aggregation of HSOBs, while CaCl2 induced both aggregation and membrane-fusion mediated coalescence of HSOBs by involving probably the anionic phospholipids together with membrane proteins. This study will contribute to extend the range of novel food products and designed emulsions containing hemp seed proteins and oil bodies.

Loading of lutein in egg-sphingomyelin vesicles as lipid carriers: Thermotropic phase behaviour, structure of sphingosome membranes and lutein crystals.

Lutein is a xanthophyll carotenoid provided exclusively by the diet, that has protective functions and beneficial effects on human health. Supplementation in lutein is necessary to reach the recommended daily dietary intake. However, the introduction of lutein into foods and beverages is a real challenge since this lipophilic nutrient has a poor aqueous solubility and a low bioavailability. In this study, we investigated the capacity of egg-sphingomyelin (ESM) vesicles called sphingosomes to solubilise lutein into the bilayers. The thermal and structural properties of ESM bilayers were examined in presence of various amounts of lutein by differential scanning calorimetry (DSC) and temperature-controlled X-ray diffraction (XRD), the structures of sphingosomes and lutein crystals were observed by microscopic techniques. ESM bilayers were in the fluid Lα phase above the phase transition temperature Tm = 39.6 °C and in the lamellar ripple Pß' phase below Tm where ESM sphingosomes exhibited ondulations and were facetted. Lutein molecules were successfully incorporated into the ESM bilayers where they induced a structural disorganisation. For ESM/lutein 90/10 %mol (91.8/8.2 %wt; 89 mg lutein / g ESM), lutein partitioning occured with the formation of lutein crystals in the aqueous phase together with lutein-loaded ESM vesicles. This study highlighted the capacity of new lipid carriers such as egg-sphingosomes to solubilise lutein and opens perspectives for the formulation of effective lutein-fortified functionnal foods and beverages providing health benefits.

Studying the real-time interplay between triglyceride digestion and lipophilic micronutrient bioaccessibility using droplet microfluidics. 2 application to various oils and (pro)vitamins.

The kinetics of micellar solubilization of lipophilic micronutrients (bioaccessibility) in relation with triglyceride digestion remains poorly known. To study this interplay in real-time, a droplet microfluidic method was designed and used as reported in the first part of this article series. In this second part, the interplay between the micellar solubilization of (pro)vitamins (beta-carotene or retinyl palmitate) and the digestion of triglyceride oils (tricaprylin TC, or high-oleic sunflower seed oil HOSO, or fish oil FO) during simulated gastrointestinal digestion was investigated. The relation between the release of both micronutrients and of triglyceride lipolytic products was found to be non-linear. The kinetics of beta-carotene was found to follow the kinetics of lipolytic products, depending on the oil type (TC > HOSO > FO). The effect of the gastric phase on the intestinal phase was also found to follow this order, mostly due to partial lipolysis during the gastric phase.

A pilot study for the intrinsic labeling of egg proteins with 15N and 13C.

The aim of this study was to produce intrinsically and uniformly doubly (15)N-(13)C-labeled proteins. These proteins can be used as intrinsic tracers of dietary amino acids, both α-amino groups and carbon skeletons, during postprandial metabolic utilization. Two (Rhodes) laying hens were fed for 16 days with a standard poultry diet supplemented with 0, 0.2% or 0.4% of a mixture of 20 doubly (15)N-(13)C-labeled AAs. A third hen was given a non-enriched diet, as the control. The eggs laid were collected over 24 days, from 3 days before to 4 days after supplementation. The (15)N and (13)C enrichments in proteins from white and yolk were measured by EA-IRMS and GC-C-IRMS for enrichment in individual amino acids. After 10 days of supplementation, the (15)N enrichment reached an isotopic plateau at 1500 to 3000 ‰, depending on the supplementation level, in both white and yolk while the (13)C enrichment was 220 to 650 ‰ in white and was 100 to 250 ‰ in yolk. The (15)N enrichment was similar among the amino acids, except for the aromatic ones in which the enrichment was lower. The δ(13)C values were variable among amino acids in both white and yolk, ranging from 77 ‰ for tyrosine to 555 ‰ for proline with the 0.2 % supplementation level. In conclusion, the incorporation of 0.2 % labeled amino acids in the hen diet allowed us to achieve sufficient enrichment for metabolic studies. However, due to the non-homogeneity of the (13)C labeling, adequate (13)C enrichment of individual amino acids must be considered depending on the investigated metabolic pathway.

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.

Caleosin of Arabidopsis thaliana : Effect of Calcium on Functional and Structural Properties.

A non-radioactive blot binding assay has proved the capacity of a purified recombinant form of Arabidopsis thaliana caleosin (AtClo1), a key protein of this plant oil body, to bind calcium. Calcium affected recombinant caleosin aggregation state, solubility, and electrophoretic mobility on SDS-PAGE. The effect of calcium on interfacial behavior of recombinant caleosin was studied at three interfaces: air/water (A/W), purified oil/water (O/W), and air/phosholipid/water (A/PLs/W). Recombinant caleosin was able to decrease interfacial tension (IFT) at A/W and O/W interfaces as a function of concentration and calcium, whereas no interaction was detected at the A/PLs/W interface. Effect of calcium was time dependent, and its amplitude strongly varied with the interface considered. Reconstituted oil bodies were used to prove the involvement of recombinant caleosin in their calcium-driven aggregation and coalescence. Calcium ions at concentration as low as 100 nM were able to strongly modify the shape and aggregation state of purified oil bodies, as well as their behavior within a monolayer, reflecting potentially profound changes in their structure and dynamic.