Project description:The possibility of stabilizing emulsions of water and non-polar alkane with pure, coloured organic pigment particles is explored. Seven pigment types each possessing a primary colour of the rainbow were selected. Their solubility in water or heptane was determined using a spectrophotometric method and their surface energies were derived from the contact angles of probe liquids on compressed disks of the particles. As expected, most of the pigments are relatively hydrophobic but pigment orange is quite hydrophilic. At equal volumes of oil and water, preferred emulsions were water-in-oil (w/o) for six pigment types and oil-in-water (o/w) for pigment orange. The emulsion type is in line with calculated contact angles of the particles at the oil-water interface being either side of 90°. Their stability to coalescence increases with particle concentration. Emulsions are shown to undergo limited coalescence from which the coverage of drop interfaces by particles has been determined. In a few cases, close-packed primary particles are visible around emulsion droplets. At constant particle concentration, the influence of the volume fraction of water (?w) on emulsions was also studied. For the most hydrophilic pigment orange, emulsions are o/w at all ?w, whereas they are w/o for the most hydrophobic pigments (red, yellow, green and blue). For pigments of intermediate hydrophobicity however (indigo and violet), catastrophic phase inversion becomes possible with emulsions inverting from w/o to o/w upon increasing ?w. For the first time, we link the pigment surface energy to the propensity of emulsions to phase invert transitionally or catastrophically.
Project description:Encapsulation of proteins can be beneficial for food and biomedical applications. To study their biophysical properties in complex coacervate core micelles (C3Ms), we previously encapsulated enhanced green fluorescent protein (EGFP) and its monomeric variant, mEGFP, with the cationic-neutral diblock copolymer poly(2-methyl-vinyl-pyridinium)n-b-poly(ethylene-oxide)m (P2MVPn-b-PEOm) as enveloping material. C3Ms with high packaging densities of fluorescent proteins (FPs) were obtained, resulting in a restricted orientational freedom of the protein molecules, influencing their structural and spectral properties. To address the generality of this behavior, we encapsulated seven FPs with P2MVP41-b-PEO205 and P2MVP128-b-PEO477. Dynamic light scattering and fluorescence correlation spectroscopy showed lower encapsulation efficiencies for members of the Anthozoa class (anFPs) than for Hydrozoa FPs derived from Aequorea victoria (avFPs). Far-UV CD spectra of the free FPs showed remarkable differences between avFPs and anFPs, caused by rounder barrel structures for avFPs and more elliptic ones for anFPs. These structural differences, along with the differences in charge distribution, might explain the variations in encapsulation efficiency between avFPs and anFPs. Furthermore, the avFPs remain monomeric in C3Ms with minor spectral and structural changes. In contrast, the encapsulation of anFPs gives rise to decreased quantum yields (monomeric Kusabira Orange 2 (mKO2) and Tag red fluorescent protein (TagRFP)) or to a pKa shift of the chromophore (FP variant mCherry).
Project description:Encapsulation of charged proteins into complex coacervate core micelles (C3Ms) can be accomplished by mixing them with oppositely charged diblock copolymers. However, these micelles tend to disintegrate at high ionic strength. Previous research showed that the addition of a homopolymer with the same charge sign as the protein improved the stability of protein-containing C3Ms. In this research, we used fluorescence correlation spectroscopy (FCS) and dynamic light scattering (DLS) to study how the addition of the homopolymer affects the encapsulation efficiency and salt stability of the micelles. We studied the encapsulation of laccase spore coat protein A (CotA), a multicopper oxidase, using a strong cationic-neutral diblock copolymer, poly(N-methyl-2-vinyl-pyridinium iodide)-block-poly(ethylene oxide) (PM2VP128-b-PEO477), and a negatively charged homopolymer, poly(4-styrenesulfonate) (PSS215). DLS indeed showed an improved stability of this three-component C3M system against the addition of salt compared to a two-component system. Remarkably, FCS showed that the release of CotA from a three-component C3M system occurred at a lower salt concentration and over a narrower concentration range than the dissociation of C3Ms. In conclusion, although the addition of the homopolymer to the system leads to micelles with a higher salt stability, CotA is excluded from the C3Ms already at lower ionic strengths because the homopolymer acts as a competitor of the enzyme for encapsulation.
Project description:Complex coacervate core micelles (C3Ms) are nanoscopic structures formed by charge interactions between oppositely charged macroions and used to encapsulate a wide variety of charged (bio)molecules. In most cases, C3Ms are in a dynamic equilibrium with their surroundings. Understanding the dynamics of molecular exchange reactions is essential as this determines the rate at which their cargo is exposed to the environment. Here, we study the molecular exchange in C3Ms by making use of Förster resonance energy transfer (FRET) and derive an analytical model to relate the experimentally observed increase in FRET efficiency to the underlying macromolecular exchange rates. We show that equilibrated C3Ms have a broad distribution of exchange rates. The overall exchange rate can be strongly increased by increasing the salt concentration. In contrast, changing the unlabeled homopolymer length does not affect the exchange of the labeled homopolymers and an increase in the micelle concentration only affects the FRET increase rate at low micelle concentrations. Together, these results suggest that the exchange of these equilibrated C3Ms occurs mainly by expulsion and insertion, where the rate-limiting step is the breaking of ionic bonds to expel the chains from the core. These are important insights to further improve the encapsulation efficiency of C3Ms.
Project description:The encapsulation of proteins into complex coacervate core micelles (C3Ms) is of potential interest for a wide range of applications. To address the stability and dynamic properties of these polyelectrolyte complexes, combinations of cyan, yellow, and blue fluorescent proteins were encapsulated with cationic-neutral diblock copolymer poly(2-methyl-vinyl-pyridinium)128- b-poly(ethylene-oxide)477. Förster resonance energy transfer (FRET) allowed us to determine the kinetics of C3M formation and of protein exchange between C3Ms. Both processes follow first-order kinetics with relaxation times of ±100 s at low ionic strength ( I = 2.5 mM). Stability studies revealed that 50% of FRET was lost at I = 20 mM, pointing to the disintegration of the C3Ms. On the basis of experimental and theoretical considerations, we propose that C3Ms relax to their final state by association and dissociation of near-neutral soluble protein-polymer complexes. To obtain protein-containing C3Ms suitable for applications, it is necessary to improve the rigidity and salt stability of these complexes.
Project description:This work aimed at studying the stabilization of O/W Pickering emulsions using nanosized cellulosic material, produced from raw cellulose or tomato pomace through different mechanical treatments, such as ball milling (BM) and high-pressure homogenization (HPH). The cellulose nanofibrils obtained via HPH, which exhibited longer fibers with higher flexibility than those obtained via ball milling, are characterized by lower interfacial tension values and higher viscosity, as well as better emulsion stabilization capability. Emulsion stability tests, carried out at 4 °C for 28 d or under centrifugation at different pH values (2.0, 7.0, and 12.0), revealed that HPH-treated cellulose limited the occurrence of coalescence phenomena and significantly slowed down gravitational separation in comparison with BM-treated cellulose. HPH-treated cellulose was responsible for the formation of a 3D network structure in the continuous phase, entrapping the oil droplets also due to the affinity with the cellulose nanofibrils, whereas BM-treated cellulose produced fibers with a more compact structure, which did adequately cover the oil droplets. HPH-treated tomato pomace gave similar results in terms of particle morphology and interfacial tension, and slightly lower emulsion stabilization capability than HPH-treated cellulose, suggesting that the used mechanical disruption process does not require cellulose isolation for its efficient defibrillation.
Project description:Few natural, biocompatible, and inexpensive emulsifiers are available because such emulsifiers must satisfy severe requirements, be produced synthetically rather than naturally, be nontoxic, and require minimal effort to produce. Therefore, the synthesis of food-grade and biocompatible nanoparticles as an alternative to surfactants has recently received attention in the industry. However, many previous efforts involved chemical modification of materials or the introduction of secondary cocomponents for emulsion formation. To achieve the goal of simple preparation, we consider here chitosan nanoparticles to prepare Pickering emulsions of food-grade oil through the control of pH, without further chemical modification or extra additives. A mild process can prepare nanoparticles from chitosan by simply increasing the pH from 3.0 to 6.0. The results showed that the average radius of chitosan at pH 6.0 was 170 nm, while large aggregates were formed at pH 6.5. These nanoparticles were utilized to prepare the Pickering emulsion. The average size of emulsion droplets decreased upon increasing the pH from 3.0 to 6.0. Moreover, Pickering emulsions at different oil fractions and nanoparticle concentrations were stable and showed a low creaming index for 45 days. The emulsions were stable against coalescence and flocculation and behaved rheologically as gel-like, shear-thinning fluids (G' > G″). Pickering emulsion prevents the growth of the microorganism (Staphylococcus aureus) at different pH values and chitosan concentrations. These results demonstrate that chitosan nanoparticles could be a cost-effective and biocompatible emulsifier for the food or pharmaceutical industry for encapsulation and bioactive compounds, and Pickering emulsions have promising antibacterial effects for further applications.
Project description:The field of biocatalysis is expanding owing to the increasing demand for efficient low-cost green chemical processes. However, a feasible strategy for achieving product separation, enzyme recovery, and high catalytic efficiency in biocatalysis remains elusive. Herein, we present thermoresponsive Pickering high internal phase emulsions (HIPEs) as controllable scaffolds for efficient biocatalysis; these HIPEs demonstrate a transition between emulsification and demulsification depending on temperature. Ultra-high-surface-area Pickering HIPEs were stabilized by Candida antarctica lipase B immobilized on starch particles modified with butyl glycidyl ether and glycidyl trimethyl ammonium chloride, thus simplifying the separation and reuse processes and significantly improving the catalytic efficiency. In addition, the switching temperature can be precisely tuned by adjusting the degree of substitution of the modified starches to meet the temperature demands of various enzymes. We believe that this system provides a green platform for various interfacial biocatalytic processes of industrial interest.
Project description:Medium and high internal phase Pickering emulsions stabilized by cellulose nanocrystals (CNCs) have been prepared and the effects of CNC concentration and type of oil phase on the properties of emulsions were studied. The maximum oil phase volume that can be stabilized by CNCs is 87% when the CNC concentration is 0.6?wt.%; this slightly decreases to 83% when the CNC concentration is increased to 1.2?wt.% or higher. In addition, the oil droplets stabilized with 0.6?wt.% CNC suspensions have a larger size than those stabilized with higher concentration CNC suspensions. As evidenced by the change in oil droplet morphology and size, two different emulsion formation mechanisms are proposed. For a CNC concentration of 0.6?wt.%, the extra oil added into the emulsion is accommodated by the expansion of oil droplet size, whereas for CNC concentrations of 1.2?wt.% and higher, the oil is stabilized mainly by the formation of new oil droplets.
Project description:Non-aqueous Pickering emulsions of 16-240 μm diameter have been prepared using diblock copolymer worms with ethylene glycol as the droplet phase and an n-alkane as the continuous phase. Initial studies using n-dodecane resulted in stable emulsions that were significantly less turbid than conventional water-in-oil emulsions. This is attributed to the rather similar refractive indices of the latter two phases. By utilizing n-tetradecane as an alternative oil that almost precisely matches the refractive index of ethylene glycol, almost isorefractive ethylene glycol-in-n-tetradecane Pickering emulsions can be prepared. The droplet diameter and transparency of such emulsions can be systematically varied by adjusting the worm copolymer concentration.