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Hypothesis Oleosomes are stabilized by a complex outer phospholipid-protein-layer. To improve understanding of its structure and stabilization mechanism, this shell has to be studied in extracellular native conditions. This should be possible by SANS using contrast variation. Oleosomes are expected to be highly temperature stable, with molecular changes occurring first in the protein shell. Direct measurements of changes in the shell structure are also important for processing methods, e.g. encapsulation. Experiments Extracted soybean oleosomes were studied directly and after encapsulation with pectin by SANS using contrast variation. In order to determine structure and size, a shell model of oleosomes was developed. The method was tested against a simple phospholipid-stabilized emulsion. The oleosomes’ temperature stability was investigated by performing SANS at elevated temperatures. Findings Size (Rg = 1380 A) and shell thickness of native and encapsulated oleosomes have been determined. This is the first report measuring the shell thickness of oleosomes directly. For native oleosomes, a shell of 9 nm thickness surrounds the oil core, corresponding to a layer of phospholipids and proteins. Up to 90 °C, no structural change was observed, confirming the oleosomes’ high temperature stability. Successful coavervation of oleosomes was shown by an increase in shell thickness of 10 nm after electrostatic deposition of pectin.

Mixed carbohydrate systems are of special interest for the food and non-food industry as they offer a versatile range of unique and novel functional properties. However, intense research is required to understand the complex processes occurring in such systems on a molecular level and to be able to modify them aim-oriented. In food, characteristic properties are based on the physicochemical functions of the biopolymers added. Thus, small deformation tests and moisture analysis have been applied to study the impact of xanthan gum and two types of sugar on the viscoelastic properties, the sol–gel transition and the water holding capacity of 1% agarose hydrogels. Agarose gels are very elastic, turbid and prone to synaeresis, which impinges on their mouth feeling. Additions of xanthan gum revealed less elastic gels with an unaffected water holding capacity. Progressive addition of two different types of up to 40% of sugar yield an increase of the elasticity of agarose gels, whereby sugar concentrations of 60% partially result in a structural breakdown and thus a significant lower network structure but better water holding. In ternary systems, the effect of the sugar concentration and sugar type used is diminished by xanthan gum. The gelation mechanism of agarose gels with a distinct amount of co-solutes is presumably mainly affected by the water shortage evolved from the competition for it of all solutes present.

Lipid storage in plants is achieved among all plant species by formation of oleosomes, enclosing oil (triacylglycerides) in small subcellular droplets. Seeds are rich in this pre-emulsified oil to provide a sufficient energy reservoir for growing. The triacylglyceride core of the oleosomes is surrounded by a phospholipid monolayer containing densely packed proteins called oleosins. They are anchored in the triacylglycerides core with a hydrophobic domain, while the hydrophilic termini remain on the surface. These specialized proteins are expressed during seed development and maturation. Particularly, they play a major role in the stabilization and function of oleosomes. To better understand the importance of oleosins for oleosome stabilization, enzymatic digestion of oleosins was performed. This made it possible to compare and correlate changes in the molecular structure of oleosins and changing macroscopic properties of oleosomes. Tryptic digestion cleaves the hydrophilic part of the oleosins, which is accompanied by a loss of secondary structures as evidenced by Fourier-transform infrared and sum frequency generation spectra. After digestion, the ability of oleosins to stabilize oil-water or air-water interfaces was lost. The surface charge and the associated aggregation behavior of oleosomes are governed by interactions typical of proteins before digestion and by interactions typical of phospholipids after digestion.

The food industry has discovered that oleosomes are beneficial as carriers of bioactive ingredients. Oleosomes are subcellular oil droplets typically found in plant seeds. Within seeds, they exist as pre-emulsified oil high in unsaturated fatty acids, stabilised by a monolayer of phospholipids and proteins, called oleosins. Oleosins are anchored into the oil core with a hydrophobic domain, while the hydrophilic domains remain on the oleosome surface. To preserve the nutritional value of the oil and the function of oleosomes, microencapsulation by means of spray drying is a promising technique. For the microencapsulation of oleosomes, maltodextrin was used. To achieve a high oil encapsulation efficiency, optimal process parameters needed to be established. In order to better understand the mechanisms of drying behind powder formation and the associated powder properties, the findings obtained using different microscopic and spectroscopic measurements were correlated with each other. By doing this, it was found that spray drying of pure oleosome emulsions resulted in excessive component segregation and thus in a poor encapsulation efficiency. With the addition of maltodextrin, the oil encapsulation efficiency was significantly improved.

The influence of the polysaccharide charge distribution on the structure, thickness, and charge reversal of the interfacial layers, formed by adsorbed positively charged protein and oppositely charged polysaccharide, has been investigated using a lattice-based self-consistent field (SCF) approach. We compare the adsorption behaviour of a uniformly charged polysaccharide model with that consisting of a short and a long block carrying different charge densities. For homogeneously charged polysaccharide we observe a resulting interfacial layer that is closer to a mixed protein + polysaccharide film, rather than a multi-layer. We also find that the maximum adsorption of polysaccharide occurs at an optimal value of its charge, above and below which the adsorbed amount decreases. In contrast, for heterogeneously charged chains, as their charge is increasingly located on the shorter block, a much thicker interfacial layer results. In such cases the weakly charged longer blocks extend well away from the surface into the solution. The interfacial film begins to resemble a multi-layer with a primary protein and a distinct secondary polysaccharide layer. When the weakly charged long blocks still have a sufficient amount of negative charge, we also observe a reversal of the sign of surface potential from a positive to a negative value. Our SCF calculated values for the reversed surface potential are of the order of −25 mV, in good agreement with several experimental results involving ζ-potential measurements on particles covered with such protein + polysaccharide films.