Designing microcapsules based on protein fibrils and protein - polysaccharide complexes

Keywords: encapsulation, microcapsule, protein, fibril, protein-polysaccharide complex, controlled release, interfacial rheology, lysozyme, ovalbumin This thesis describes the design of encapsulation systems using mesostructures from proteins and polysaccharides. The approach was to first investigate the physical properties of the encapsulating materials (protein fibrils and protein – polysaccharide complexes). Subsequently, microcapsules with tunable release rate and mechanical strength were developed. Firstly, the effect of steady shear and turbulent flow on the formation of protein fibrils from lysozyme was studied. We determined the conversion and size distribution of fibrils obtained by heating lysozyme solutions at pH 2. The formation of fibrils was quantified using flow-induced birefringence. The size distribution was fitted using decay of birefringence measurements and Transmission Electron Microscopy. The morphology of Lys fibrils and kinetics of their formation varied considerably depending on the flow applied. With increasing shear or stirring rate, more rod-like and shorter fibrils were obtained, and the conversion into fibrils was increased. Secondly, we have investigated the surface rheological properties of oil – water interfaces stabilized by fibrils from lysozyme (long and semi-flexible, and short and rigid ones), fibrils from ovalbumin (short and semi-flexible), lysozyme – pectin complexes, or ovalbumin – pectin complexes. We have compared these properties with those of interfaces stabilized by the native proteins. The surface dilatational and surface shear moduli were determined using an automated drop tensiometer, and a stress controlled rheometer with biconical disk geometry. Results show that interfaces stabilized by protein – pectin complexes have higher surface shear and dilatational moduli than interfaces stabilized by the native proteins only. At most of the experimental conditions, interfaces stabilized by protein fibrils have the highest surface rheological moduli. The difference between long semi-flexible lysozyme fibrils or short rigid lysozyme fibrils is not pronounced in interfacial dilation rheology but significant in interfacial shear rheology. The complex surface shear moduli of interfaces stabilized by long semi-flexible fibrils are about ten times higher than those of interfaces stabilized by short rigid fibrils, over a range of bulk concentrations. Interfaces stabilized by short and more flexible ovalbumin fibrils have a significantly higher surface shear modulus than those stabilized by the somewhat longer and more rigid short lysozyme fibrils. Finally, encapsulation systems are developed using layer-by-layer adsorption of food-grade polyelectrolytes on an emulsion droplet template. The first encapsulation system was built with alternating layers of ovalbumin fibrils and high methoxyl pectin. By varying the number of layers, the release of active ingredients can be controlled: increasing the number of layers of the shell from four to eight, decreases the release rate by a factor six. The other encapsulation systems were built with alternating layers of protein – pectin complexes and protein fibrils. Two types of proteins (ovalbumin and lysozyme) and three types of fibrils were used: short and semi-flexible from ovalbumin, short and rod-like, and long and semi-flexible from lysozyme. At low number of layers (less than five), microcapsules from ovalbumin complexes and fibrils were stronger than microcapsules prepared from lysozyme complexes and fibrils. Increasing the number of layers, the mechanical stability of microcapsules from lysozyme complexes and fibrils increased significantly, and capsules were stronger than those prepared from ovalbumin complexes and fibrils with the same number of layers. The contour length of the Lys fibrils did not have a significant effect on mechanical stability of the lysozyme complexes and fibrils capsules. These results show that mechanical properties of this type of capsule can be tuned by varying the flexibility of the protein fibrils. </p

Mealworm Products with Custom-made Texture

Due to the environmental impact of animal meat such as beef, pork and poultry, a need for alternative protein sources and meat substitutes is growing. Insects seem to provide the optimal alternative

Nonlinear stress deformation behavior of interfaces stabilized by food-based ingredients

Interfaces stabilized by food-based ingredients, such as proteins or glycolipids, often display nonlinear behavior when subjected to oscillatory dilatational deformations, even at the lowest deformation amplitudes which can currently be applied experimentally. Here we show that classical approaches to extract dilatational properties, based on the Young–Laplace equation, may not always be suitable to analyze data. We discuss a number of examples of foodingredient stabilized interfaces (interfaces stabilized by protein fibrils, protein–polysaccharide complexes and oligosaccharide–fatty aid conjugates) and show how an analysis of the dynamic surface tension signal using Lissajous plots and a protocol which includes deformation amplitude and droplet size variations, can be used to obtain a more detailed and accurate description of their nonlinear dilatational behavior

Surface rheological properties of liquid-liquid interfaces stabilized by protein fibrillar aggregates and protein-polysaccharide complexes

In this study we have investigated the surface rheological properties of oil-water interfaces stabilized by fibrils from lysozyme (long and semi-flexible and short and rigid ones), fibrils from ovalbumin (short and semi-flexible), lysozyme-pectin complexes, or ovalbumin-pectin complexes. We have compared these properties with those of interfaces stabilized by the native proteins. The surface dilatational and surface shear moduli were determined using an automated drop tensiometer and a stress controlled rheometer with biconical disk geometry. Results show that interfaces stabilized by complexes of these proteins with high-methoxyl pectin have higher surface shear and dilatational moduli than interfaces stabilized by the native proteins only. The interfaces stabilized by ovalbumin and lysozyme complexes have comparable shear and dilatational moduli though ovalbumin-pectin complexes are twice as large in radius as lysozyme-pectin complexes. Under most of the experimental conditions, interfaces stabilized by fibrils have the highest surface rheological moduli. The difference between long semi-flexible lysozyme fibrils or short rigid lysozyme fibrils is not pronounced in interfacial dilation rheology but significant in interfacial shear rheology. The complex surface shear moduli of interfaces stabilized by long semi-flexible fibrils are about 10 times higher than those of interfaces stabilized by short rigid fibrils, over a range of bulk concentrations. Interfaces stabilized by short and more flexible ovalbumin fibrils have a significantly higher surface shear modulus than those stabilized by longer and more rigid lysozyme fibrils. This study has shown that the use of such supra-molecular structural building blocks creates a wider range of microstructural features of the interface, with higher surface shear and dilatational moduli and a more complex dependence on strain

Mealworm Products with Custom-made Texture

Due to the environmental impact of animal meat such as beef, pork and poultry, a need for alternative protein sources and meat substitutes is growing. Insects seem to provide the optimal alternative

Surface rheological properties of liquid-liquid interfaces stabilized by protein fibrillar aggregates and protein-polysaccharide complexes

In this study we have investigated the surface rheological properties of oil-water interfaces stabilized by fibrils from lysozyme (long and semi-flexible and short and rigid ones), fibrils from ovalbumin (short and semi-flexible), lysozyme-pectin complexes, or ovalbumin-pectin complexes. We have compared these properties with those of interfaces stabilized by the native proteins. The surface dilatational and surface shear moduli were determined using an automated drop tensiometer and a stress controlled rheometer with biconical disk geometry. Results show that interfaces stabilized by complexes of these proteins with high-methoxyl pectin have higher surface shear and dilatational moduli than interfaces stabilized by the native proteins only. The interfaces stabilized by ovalbumin and lysozyme complexes have comparable shear and dilatational moduli though ovalbumin-pectin complexes are twice as large in radius as lysozyme-pectin complexes. Under most of the experimental conditions, interfaces stabilized by fibrils have the highest surface rheological moduli. The difference between long semi-flexible lysozyme fibrils or short rigid lysozyme fibrils is not pronounced in interfacial dilation rheology but significant in interfacial shear rheology. The complex surface shear moduli of interfaces stabilized by long semi-flexible fibrils are about 10 times higher than those of interfaces stabilized by short rigid fibrils, over a range of bulk concentrations. Interfaces stabilized by short and more flexible ovalbumin fibrils have a significantly higher surface shear modulus than those stabilized by longer and more rigid lysozyme fibrils. This study has shown that the use of such supra-molecular structural building blocks creates a wider range of microstructural features of the interface, with higher surface shear and dilatational moduli and a more complex dependence on strain

Effects of Flow on Hen Egg White Lysozyme (HEWL) Fibril Formation: Lenght Distribution, Flexibility, and Kinetics

The effect of steady shear and turbulent flow on the formation of amyloid fibrils from hen egg white lysozyme (HEWL) was studied. The conversion and size distribution of fibrils obtained by heating HEWL solutions at pH 2 were determined. The formation of fibrils was quantified using flow-induced birefringence. The size distribution was fitted using decay of birefringence measurements and transmission electron microscopy (TEM). The morphology of HEWL fibrils and the kinetics of their formation varied considerably depending on the flow applied. With increasing shear or stirring rate, more rod-like and shorter fibrils were obtained, and the conversion into fibrils was increased. The size distribution and final fibril concentration were significantly different from those obtained in the same heat treatment at rest. The width of the length distribution of fibrils was influenced by the homogeneity of the flow

Microcapsules with Protein Fibril Reinforced Shells: Effect of Fibril Properties on Mechanical Strength of the Shell

In this study, we produced microcapsules using layer-by-layer adsorption of food-grade polyelectrolytes on an emulsion droplet template. We compared the mechanical stability of microcapsules to shells consisting of alternating layers of ovalbumin–high methoxyl pectin (Ova–HMP) complexes and semi-flexible ovalbumin (Ova) fibrils (average contour length, Lc 200 nm), with microcapsules built of alternating layers of lysozyme–high methoxyl pectin (LYS–HMP) complexes and lysozyme (LYS) fibrils. Two types of LYS fibrils were used: short and rod-like (Lc 500 nm) and long and semi-flexible (Lc = 1.2–1.5 µm). At a low number of layers (=4), microcapsules from Ova complexes and fibrils were stronger than microcapsules prepared from LYS complexes and fibrils. With an increase of the number of layers, the mechanical stability of microcapsules from LYS–HMP/LYS fibrils increased significantly and capsules were stronger than those prepared from Ova–HMP/Ova fibrils with the same number of layers. The contour length of the LYS fibrils did not have a significant effect on mechanical stability of the LYS–HMP/LYS fibril capsules. The stiffer LYS fibrils produce capsules with a hard but more brittle shell, whereas the semi-flexible Ova fibrils produce capsules with a softer but more stretchable shell. These results show that mechanical properties of this type of capsule can be tuned by varying the flexibility of the protein fibrils

Effects of Flow on Hen Egg White Lysozyme (HEWL) Fibril Formation: Lenght Distribution, Flexibility, and Kinetics

The effect of steady shear and turbulent flow on the formation of amyloid fibrils from hen egg white lysozyme (HEWL) was studied. The conversion and size distribution of fibrils obtained by heating HEWL solutions at pH 2 were determined. The formation of fibrils was quantified using flow-induced birefringence. The size distribution was fitted using decay of birefringence measurements and transmission electron microscopy (TEM). The morphology of HEWL fibrils and the kinetics of their formation varied considerably depending on the flow applied. With increasing shear or stirring rate, more rod-like and shorter fibrils were obtained, and the conversion into fibrils was increased. The size distribution and final fibril concentration were significantly different from those obtained in the same heat treatment at rest. The width of the length distribution of fibrils was influenced by the homogeneity of the flow

Impact of processing on enzymatic browning and texturization of yellow mealworms

The potential of Tenebrio molitor as an alternative protein source for food applications was investigated by using several conventional (blanching and freeze drying) and non-conventional (high pressure) processing techniques. Some of the challenges in product development are the browning effect of minced mealworms and the texturizing capacity after mincing. Both blanching and high pressure processing at 400 and 500 MPa proved to be effective in inactivating enzymes responsible for browning. However, the inactivation process was accompanied by a significant decrease in texturizing properties of the minced mealworms, especially when blanching was applied. Also when applying pressure, the texture changed from stronger (samples treated at lower pressures) to weaker (samples treated at higher pressures). Minced fresh or freeze dried mealworms provided textures with a reasonable strength and elasticity, but browning still occurred. Nevertheless, this browning could be slowed down by applying air tight packaging to prevent oxidation.</p