Emulsions Stabilized with Polyelectrolyte Complexes Prepared from a Mixture of a Weak and a Strong Polyelectrolyte.

The possibility of stabilizing emulsions with polyelectrolyte complexes (PEC) obtained from the interaction of two non-surface-active oppositely charged polyelectrolytes (PEL) is described. Poly(allylamine hydrochloride) (PAH) and poly(4-styrene sulfonate) sodium salt are selected as the weak cationic and the strong anionic polyelectrolyte, respectively. Aqueous polymer mixtures are investigated by light scattering to determine the size of the complexes and whether precipitation or complex coacervation occurs. The effects of PEL mixing ratio, pH, and PEL concentration are studied in detail. By increasing the pH, the transition precipitate-precipitate/coacervate-coacervate-polymer solution is observed. At low pH, both PEL are fully ionized and therefore precipitates (soft particles) arise as a result of strong electrostatic interactions. By increasing the pH, the degree of ionization of PAH decreases and weak electrostatic interactions ensue, supporting the formation of coacervate droplets. The most stable oil-in-water emulsions are prepared from aqueous mixtures around charge neutralization. Although emulsions can be prepared from coacervate droplet dispersions, their coalescence stability is worse than those stabilized by soft PEC particles. By increasing the PEL concentration, the average droplet diameter decreases and the fraction of cream in the emulsion increases for emulsions prepared with PEC particles, following the limited coalescence model. However, at high concentrations, emulsion stability is slightly worse probably due to extensive aggregation of the particles. Viscous high internal phase emulsions can be prepared at low pH in which oil droplets are deformed. Here, PEC particles are detected only at the oil-water interface. At lower oil content, excess particles form a network in the aqueous phase aiding emulsion stability to coalescence.

Emulsion stabilisation by complexes of oppositely charged synthetic polyelectrolytes.

We investigate the possibility of stabilising oil-water emulsions from the polyelectrolyte complexes (PEC) obtained in mixtures of a strong cationic polyelectrolyte (poly(diallyldimethylammonium chloride), PDADMAC) and a weak anionic one (poly(acrylic acid)sodium salt, PAANa). Unlike other previous work however, both polyelectrolytes (PEL) are chosen as they are completely water-soluble and possess no surface activity when present alone over nearly all the pH range. In water, the effects of PEL concentration, PEL mixing ratio and pH on the formation of PEC are studied in detail. At low pH where the anionic PEL is uncharged, complex coacervation occurs in which droplets rich in both polymers are dispersed in water. At intermediate pH, the PEC comprise a mixture of coacervate droplets and solid particles. At high pH where the anionic PEL is significantly charged, only complex coacervation is observed. On addition of dodecane followed by homogenisation, no stable emulsions arose from dispersions containing solid particle PEC due to either the large precursor particle aggregates or their inherent hydrophilicity. By contrast, oil-in-water emulsions stable to coalescence could be prepared from coacervate dispersions. We discuss the feasibility of the coacervate phase spreading at the oil-water interface in terms of the relevant spreading coefficients and compare the predictions with experiment for a range of oils. We encounter oils whose drops become engulfed by the coacervate phase as well as oils where no engulfing occurs.

Novel stabilisation of emulsions by soft particles: polyelectrolyte complexes.

We put forward the concept of a novel particle stabiliser of oil-water emulsions, being the polyelectrolyte complex (PEC) formed between oppositely charged water-soluble polymers in cases where either polymer alone is incapable of stabilising an emulsion. Using poly(4-styrene sulfonate) sodium salt, PSSNa and poly(diallyldimethylammonium chloride), PDADMAC, of low polydispersity and similar molecular mass, we correlate the behaviour of their mixtures in water with that of emulsions after addition of oil. In aqueous mixtures, spherical particles of diameters between 100 and 150 nm are formed through electrostatic interactions between charged polymer chains. Around equal mole fractions of the two polymers, the zeta potential of the particles reverses in sign and emulsions of oil-in-water (o/w) for a range of oils can be prepared which are the most stable to coalescence and creaming. The effects of PEC concentration and the oil : water ratio have been examined. All emulsions are o/w and stability is achieved by close-packed particle layers at drop interfaces and particle aggregation in the continuous phase. Increasing the salt concentration initially causes destabilisation of the aqueous particle dispersion due to particle aggregation followed by dissolution of particles at high concentrations; the corresponding emulsions change from being stable to completely unstable and are then re-stabilised due to adsorption of uncharged individual polymer molecules.

Particles at Oil-Air Surfaces: Powdered Oil, Liquid Oil Marbles, and Oil Foam.

The type of material stabilized by four kinds of fluorinated particles (sericite and bentonite platelet clays and spherical zinc oxide) in air-oil mixtures has been investigated. It depends on the particle wettability and the degree of shear. Upon vigorous agitation, oil dispersions are formed in all the oils containing relatively large bentonite particles and in oils of relatively low surface tension (γla < 26 mN m(-1)) like dodecane, 20 cS silicone, and cyclomethicone containing the other fluorinated particles. Particle-stabilized oil foams were obtained in oils having γla > 26 mN m(-1) where the advancing air-oil-solid contact angle θ lies between ca. 90° and 120°. Gentle shaking, however, gives oil-in-air liquid marbles with all the oil-particle systems except for cases where θ is <60°. For oils of tension >24 mN m(-1) with omniphobic zinc oxide and sericite particles for which advancing θ ≥ 90°, dry oil powders consisting of oil drops in air which do not leak oil could be made upon gentle agitation up to a critical oil:particle ratio (COPR). Above the COPR, catastrophic phase inversion of the dry oil powders to air-in-oil foams was observed. When sheared on a substrate, the dry oil powders containing at least 60 wt % of oil release the encapsulated oil, making these materials attractive formulations in the cosmetic and food industries.

A macroscopic reaction: direct covalent bond formation between materials using a Suzuki-Miyaura cross-coupling reaction.

Cross-coupling reactions are important to form C-C covalent bonds using metal catalysts. Although many different cross-coupling reactions have been developed and applied to synthesize complex molecules or polymers (macromolecules), if cross-coupling reactions are realized in the macroscopic real world, the scope of materials should be dramatically broadened. Here, Suzuki-Miyaura coupling reactions are realized between macroscopic objects. When acrylamide gel modified with an iodophenyl group (I-gel) reacts with a gel possessing a phenylboronic group (PB-gel) using a palladium catalyst, the gels bond to form a single object. This concept can also be adapted for bonding between soft and hard materials. I-gel or PB-gel selectively bonds to the glass substrates whose surfaces are modified with an electrophile or nucleophile, respectively.

Supramolecular adhesives to hard surfaces: adhesion between host hydrogels and guest glass substrates through molecular recognition.

Supramolecular materials based on host-guest interactions should exhibit high selectivity and external stimuli-responsiveness. Among various stimuli, redox and photo stimuli are useful for its wide application. An external stimuli-responsive adhesive system between CD host-gels (CD gels) and guest molecules modified glass substrates (guest Sub) is focused. Here, the selective adhesion between host gels and guest substrates where adhesion depends on molecular complementarity is reported. Initially, it is thought that adhesion of a gel material onto a hard material might be difficult unless many guest molecules modified linear polymers immobilize on the surface of hard materials. However, reversible adhesion of the CD gels is observed by dissociating and re-forming inclusion complex in response to redox and photo stimuli.

Dry oil powders and oil foams stabilised by fluorinated clay platelet particles.

A series of platelet sericite particles coated to different extents with a fluorinating agent has been characterised and their behaviour in mixtures with air and oil studied. The material which forms by vigorous shaking depends on both the surface tension of the oil and the surface energy of the particles which control their degree of wetting. Oil dispersions are formed in liquids of relatively low tension (<22 mN m(-1)), e.g. hexane and cyclomethicone, for all particles. Particle-stabilised air-in-oil foams form in liquids of higher tension, e.g. dodecane and phenyl silicone, where the advancing three-phase contact angle θ, measured on a planar substrate composed of the particles into the liquid, lies between ca. 65° and 120°. For oils of tension above 27 mN m(-1) like squalane and liquid paraffin with particles for which θ > 70°, we have discovered that dry oil powders in which oil drops stabilised by particles dispersed in air (oil-in-air) can be prepared by gentle mixing up to a critical oil : particle ratio (COPR) and do not leak oil. These powders, containing up to 80 wt% oil, release the encapsulated oil when sheared on a substrate. For many of the systems forming oil powders, stable liquid oil marbles can also be prepared. Above the COPR, catastrophic phase inversion occurs yielding an ultra-stable air-in-oil foam. We thus demonstrate the ability to disperse oil drops or air bubbles coated with particles within novel materials.

Pulsed-field gradient NMR measurements on hydrogels from phosphocholine.

Gels from diacylphosphatidylcholine in glycerol/butylene glycol mixtures were investigated by pulsed-field gradient NMR measurements. Previous measurements had shown that the gels are formed by networks from crystalline multilamellar vesicles (MLV). The obtained self-diffusion coefficients for water and butylene glycol molecules indicate that both molecules occur in two different environments, even at temperatures above the phase transition T(m) where the system is still in a liquid crystalline state. While the larger fraction of the molecules shows a free self-diffusion process like in a homogeneous phase, the smaller fraction seems to be encapsulated in closed domains and undergoes only hindered self-diffusion. It is concluded that the hindered diffusions are due to the solvent molecules trapped between the bilayers of the multilamellar vesicles, while the free diffusion is assigned to the solvent molecules outside of the MLV. Since the fraction of the entrapped molecules does not change during phase transition, we assume that the structure of the network in the samples remains the same when gelation occurs. The gelation process is simply due to the transformation of the vesicle bilayers from the liquid crystalline to the crystalline state. The permeability of the bilayer for the solvent molecules is drastically changed by this transition. The exchange of water molecules through the bilayers slows down significantly below T(m): while the average residence time of water molecules inside the vesicles is smaller than 50 ms in the liquid crystalline state, this value increases to more than 1 s for the gel state. In the case of pure butylene glycol, no vesicles are present, and it is likely that these gels are formed from crystalline fibers.

Desensitization of feedback inhibition of the Saccharomyces cerevisiae gamma-glutamyl kinase enhances proline accumulation and freezing tolerance.

In response to osmotic stress, proline is accumulated in many bacterial and plant cells as an osmoprotectant. The yeast Saccharomyces cerevisiae induces trehalose or glycerol synthesis but does not increase intracellular proline levels during various stresses. Using a proline-accumulating mutant, we previously found that proline protects yeast cells from damage by freezing, oxidative, or ethanol stress. This mutant was recently shown to carry an allele of PRO1 which encodes the Asp154Asn mutant gamma-glutamyl kinase (GK), the first enzyme of the proline biosynthetic pathway. Here, enzymatic analysis of recombinant proteins revealed that the GK activity of S. cerevisiae is subject to feedback inhibition by proline. The Asp154Asn mutant was less sensitive to feedback inhibition than wild-type GK, leading to proline accumulation. To improve the enzymatic properties of GK, PCR random mutagenesis in PRO1 was employed. The mutagenized plasmid library was introduced into an S. cerevisiae non-proline-utilizing strain, and proline-overproducing mutants were selected on minimal medium containing the toxic proline analogue azetidine-2-carboxylic acid. We successfully isolated several mutant GKs that, due to extreme desensitization to inhibition, enhanced the ability to synthesize proline better than the Asp154Asn mutant. The amino acid changes were localized at the region between positions 142 and 154, probably on the molecular surface, suggesting that this region is involved in allosteric regulation. Furthermore, we found that yeast cells expressing Ile150Thr and Asn142Asp/Ile166Val mutant GKs were more tolerant to freezing stress than cells expressing the Asp154Asn mutant.

The production of recombinant APRP, an alkaline protease derived from Bacillus pumilus TYO-67, by in vitro refolding of pro-enzyme fixed on a solid surface.

Bacillus pumilus TYO-67 has been isolated from tofuyo, a traditional fermented food made from soybean milk in Okinawa, Japan. This bacterium secretes a soybean-milk-coagulating enzyme (SMCE), which can be applied for the production of processed foods from soybean milk. Thus, an easy method of producing the recombinant enzyme was developed in this study. SMCE is an alkaline serine protease belonging to the subtilisin family; its candidate gene, aprP, which encodes a prepro-enzyme, was isolated in a previous study. Recombinant APRP was then produced by in vitro refolding of pro-APRP-His, i.e., N-terminally His-tagged pro-APRP. A large amount of pro-APRP-His was produced in Esherichia coli BL21(DE3) (ca. 8 mg from a 20-ml culture), collected as insoluble protein, dissolved in 6 M guanidine-HCl (pH 8.0), bound to Ni-NTA, and refolded on the resin at pH 10.0 to become mature APRP by autocleavage. Then, 0.16 mg of purified mature APRP was obtained through single-step chromatography from the refolded sample using 10 mg of pro-APRP-His. The N-terminal sequence and the enzymatic properties of refolded APRP were identical to those of SMCE. In addition, the pro-sequence was found to be essential for the production of mature APRP, suggesting that it could function as an intramolecular chaperone.