Enzymatically synthesized dextran nanoparticles and their use as carriers for nutraceuticals.

This study evaluated the use of enzymatically synthesized dextran nanoparticles to entrap a hydrophobic nutraceutical, the isoflavone genistein. The focus was on the synthesis of pure dextran as the material for the entire nanoparticle, and their inclusion-complexation of genistein. Under optimal conditions (pH 5.2-6 and sucrose concentration >0.5 M), dextransucrase generated spherical dextran nanoparticles (100-450 nm). The two nutraceutical inclusion methods were DMSO dilution in water and acidification. Optimization of the inclusion processes produced nanosized dextran particles containing genistein. The DMSO method was found to be more suitable for inclusion of genistein in dextran, resulting in a higher genistein load (5.6 ± 0.1 g genistein per 100 g particles), and a higher percentage of nanosized particles (85%, 105-400 nm). For both protocols, addition of a freeze-drying step exerted a positive effect presumably due to the formation of new hydrogen bonds and van der Waals interactions.

Flavonoids as dietary regulators of nuclear receptor activity.

Metabolic diseases such as obesity, type II diabetes, and dyslipidemia are a rising cause of mortality worldwide. The progression of many metabolic diseases is fundamentally regulated on the transcriptional level by a family of ligand-activated transcription factors, called nuclear receptors, which detect and respond to metabolic changes. Their role in maintaining metabolic homeostasis makes nuclear receptors an important pharmaceutical and dietary target. This review will present the growing evidence that flavonoids, natural secondary plant metabolites, are important regulators of nuclear receptor activity. Structural similarities between flavonoids and cholesterol derivatives combined with the promiscuous nature of most nuclear receptors provide a wealth of possibilities for pharmaceutical and dietary modulation of metabolism. While the challenges of bringing flavonoid-derived therapeutics to the market are significant, we consider this rapidly growing field to be an essential aspect of the functional food initiative and an important mine for pharmaceutical compounds.

Mixed micellization between natural and synthetic block copolymers: ß-casein and Lutrol F-127.

Amphiphilic block copolymers and mixtures of amphiphiles find broad applications in numerous technologies, including pharma, food, cosmetic and detergency. Here we report on the interactions between a biological charged diblock copolymer, ß-casein, and a synthetic uncharged triblock copolymer, Lutrol F-127 (EO(101)PO(56)EO(101)), on their mixed micellization characteristics and the micelles' structure and morphology. Isothermal titration calorimetry (ITC) experiments indicate that mixed micelles form when Lutrol is added to monomeric as well as to assembled ß-casein. The main driving force for the mixed micellization is the hydrophobic interactions. Above ß-casein CMC, strong perturbations caused by penetration of the hydrophobic oxypropylene sections of Lutrol into the protein micellar core lead to disintegration of the micelles and reformation of mixed Lutrol/ß-casein micelles. The negative enthalpy of micelle formation (ΔH) and cooperativity increase with raising ß-casein concentration in solution. ζ-potential measurements show that Lutrol interacts with the protein micelles to form mixed micelles even below its critical micellization temperature (CMT). They further indicate that Lutrol effectively masks the protein charges, probably by forming a coating layer of the ethyleneoxide rich chains. Small-angle X-ray scattering (SAXS) and cryogenic-transmission electron microscopy (cryo-TEM) indicate relatively small changes in the oblate micellar shape, but do show swelling along the small axis of ß-casein micelles in the presence of Lutrol, thereby confirming the formation of mixed micelles.

Effect of Hofmeister anions on micellization and micellar growth of the surfactant cetylpyridinium chloride.

Controlling the morphological characteristics of micellar solutions is important for surfactant performance and for achieving desired properties. In this work we study how monovalent anions of the lyotropic series affect micellization, micellar transitions, and micellar growth of the cationic surfactant N-cetyl pyridinium chloride (CPyCl), with the aim of achieving a tool to methodically tune these self-assembly characteristics. For the first time, a set of ions of the Hofmeister series were studied by combining indirect (surface tension, conductivity, optical absorption, viscosity, dynamic light scattering) and direct-imaging cryogenic-transmission electron microscopy (cryo-TEM). Following recent literature on anionic surfactants, we considered the pyridinium headgroup as a chaotropic cation, interacting with cosmotrope and chaotrope anions (Cl(-), Br(-), NO(3)(-), ClO(3)(-)). We show that the micelles' structure is strongly influenced by both the nature and concentration of added anions and their location in the lyotropic series, but the lyotropic number by itself cannot explain all the effects measured. Especially interesting was the relatively small effect of the chlorate ion on the CMC, but its large effect on micellar transition and growth. We further test the influence of a hydrotrope on the first and second CMC and micellar growth, and compare it with the data obtained with the inorganic salts.

Origins of the viscosity peak in wormlike micellar solutions. 1. Mixed catanionic surfactants. A cryo-transmission electron microscopy study.

The rheology of wormlike micelles ("worms") formed by surfactants in water often follows nonmonotonic trends as functions of composition. For example, a study by Raghavan et al. (Langmuir 2002, 18, 3797) on mixtures of the anionic surfactant sodium oleate (NaOA) and the cationic surfactant octyl trimethylammonium bromide (OTAB) reported a pronounced peak in the zero-shear viscosity eta0 as a function of NaOA/OTAB ratio at a constant surfactant concentration (3 wt %). In this work, we study the origins of rheological changes in the NaOA/OTAB system and the relations between the composition and structural characteristics using cryo-transmission electron microscopy (cryo-TEM). When either surfactant is in large excess, the dominating morphology is that of spherical micelles. As oppositely charged surfactant is added to the mixture, the spheres grow into linear worms and these continue to elongate as the viscosity peak (which occurs at a 70/30 NaOA/OTAB ratio) is approached from either end. At the viscosity peak, the sample shows numerous long worms as well as a small number of branched worms. Taken together, NaOA/OTAB rheology can be primarily understood on the basis of micellar growth, which is explained primarily by packing arguments. While the size of the hydrophobic micellar core continuously decreases as the short amphiphile OTAB is added at the expense of NaOA, screening of charges goes through a maximum, which contributes to the asymmetry of the viscosity curve. With regard to micellar branching, there is no significant difference in the density of branched worms on either side of the viscosity peak. Therefore, it appears that in contrast to the behavior of some surfactant/salt systems, branching does not have a significant influence on the rheology of this mixed catanionic surfactant system. Instead, our data clearly indicate that the origin of the viscosity peak is linked with micellar growth and micellar shortening.

Effect of temperature on self-assembly of bovine beta-casein above and below isoelectric pH. Structural analysis by cryogenic-transmission electron microscopy and small-angle X-ray scattering.

beta-Casein is one of the main proteins in milk, recently classified as an intrinsically unstructured protein. At neutral pH, it is composed of a highly polar N-terminus domain and a hydrophobic C-terminus tail. This amphiphilic block-copolymer-like structure leads to self-organization of the protein monomers into defined micelles. Recently, it has been shown that at room temperature, beta-casein also self-organizes into micelles in an acidic environment, but the effect of temperature on the micelles' formation and properties at the low pH regime were not explored. In the present study, we used two complementary techniques, cryogenic-transmission electron microscopy (cryo-TEM) and small-angle X-ray scattering (SAXS), to characterize at high-resolution the micelles' shape, dimensions, and aggregation numbers and to determine how these properties are affected by temperature between 1 and 40 degrees C. Two different regimes were studied: highly acidic pH where the protein is cationic, and neutral pH, where it is anionic. We found that flat disk-like micelles with low aggregation numbers formed at low temperature in the two pH regimes. Close to neutral pH increase in temperature involves a transition in the micelles' shape and dimensions from flat disks to bulky, almost spheroidal micelles, coupled with a sharp increase in the micelles' aggregation number. In contrast, no effects on the micelles' morphology or aggregation number were detected in the acidic environment within the entire temperature range studied. The self-organization into disk micelles and the lack of effect of temperature in the acidic environment are linked to the unstructured character of the protein and to the charge distribution map. The latter indicates that below the isoelectric pH (pI), beta-casein loses the distinct separation of hydrophobic and hydrophilic domains, thereby suggesting that it may no longer be considered as a classical head-tail block-copolymer amphiphile as in neutral pH.

Self-assembly of bovine beta-casein below the isoelectric pH.

Beta-casein is an intrinsically unstructured amphiphilic protein that self-assembles into micelles at neutral pH. This paper reports that beta-casein self-organizes into micelles also under acidic conditions. The protein association behavior and micelle characteristics at pH 2.6, well below the p I, are presented. The pH was found to strongly affect the micelle shape and dimensions. Cryogenic transmission electron microscopy (cryo-TEM) experiments revealed disk-like micelles of 20-25 nm in length and approximately 3.5 nm in height in acidic conditions. An aggregation number of 6 was determined by sedimentation equilibrium under these conditions. Isothermal titration calorimetry experiments verified the association below the p I and allowed determination of the micellization enthalpy, the critical micellar concentration, and the micellization relative cooperativity (MR). Small-angle X-ray scattering results at concentrations below the critical micellization concentration (CMC) suggest that the monomeric protein is likely in a premolten globule state at low pH. Calculations of the protein charge at acidic and neutral pH reveal a similar high net charge but considerable differences in the charge distribution along the protein backbone. Overall the results show that beta-casein is amphiphilic at low pH, but the distribution of charge along the protein chain creates packing constraints that affect the micelle organization, leading at concentrations above the CMC to the formation of disk micelles.

Mechanism of crumb toughening in bread-like products by microwave reheating.

Comparing breads reheated in conventional and microwave ovens revealed that the latter considerably toughens the crumb texture when internal boiling is induced. Moisture loss in itself has a relatively minor toughening effect. The major changes, caused by boiling, occur only in systems with starch concentration in excess of a threshold level of about 37% (wet basis). Substantially greater amounts of amylose are leached out of the granules in the case of sustained boiling during microwave heating, as compared to conventional oven heating. The free amylose solution is being "pushed" by the generated steam pressure toward the air-cell wall interface. A rich amylose phase is accumulated at that interface and over the granules. Upon cooling, the amylose undergoes rapid phase changes; thus, toughening is apparent in a relatively short time after heating. Minimizing the textural deleterious effects in microwave reheating of bread-like products should entail (a) preventing or minimizing internal boiling, (b) diluting of the starch concentration below the threshold level, (c) interfering with the amylose phase change by using complex forming agents.

Micellization of bovine beta-casein studied by isothermal titration microcalorimetry and cryogenic transmission electron microscopy.

The association behavior, critical micellization concentration (CMC), and enthalpy of demicellization (DeltaHdemic) of bovine beta-casein were studied, for the first time by isothermal titration calorimetry, in a pH 7.0 phosphate buffer with 0.1 ionic strength and in pure water. In the buffer solutions, the CMC decreased asymptotically from 0.15 to 0.006 mM as the temperature was raised from 16 to 45 degrees C. DeltaHdemic decreased with increasing temperature between 16 and 28 degrees C but increased from 28 to 45 degrees C. Thermodynamic analysis below 30 degrees C is consistent with the Kegeles shell model, which suggests a stepwise association process. At higher temperatures, this model exhibits limitations, and the micellization becomes much more cooperative. The CMC values in water, measured between 17 and 28 degrees C, decreased with increasing temperature and, expectedly, were higher than those found in the buffer solutions. beta-Casein micelles were visualized and characterized, for the first time in their hydrated state, using advanced digital-imaging cryogenic transmission electron microscopy. The images revealed small, oblate micelles, about approximately 13 nm in diameter. The micelles shape and dimensions remained nearly constant in the temperature range of 24-35 degrees C.