Reversible precipitation of casein micelles with a cationic hydroxyethylcellulose

The cationic hydroxyethylcellulose Polyquaternium 10 (PQ10) was found to produce a dose-dependent destabilization of casein micelles from whole or skim milk without affecting the stability of most of the whey proteins. The anionic phosphate residues on caseins were not determinant in the observed interaction since the destabilization was also observed with dephosphorylated caseins to the same extent. However, the precipitation process was completely inhibited by rising NaCl concentration, indicating an important role of electrostatic interactions. Furthermore, the addition of 150 mM NaCl solubilized preformed PQ10-casein complexes, rendering a stable casein suspension without a disruption of the internal micellar structure as determined by dynamic light scattering. This casein preparation was found to contain most of the Ca2+ and only 10% of the lactose originally present in milk and remained as a stable suspension for at least 4 months at 4 degrees C. The final concentration of PQ10 determined both the size of the casein-polymer aggregates and the amount of milkfat that coprecipitates. The presence of PQ10 in the aggregates did not inhibit the activity of rennet or gastrointestinal proteases and lipases, nor did it affect the growth of several fermentative bacteria. The cationic cellulose PQ10 may cause a reversible electrostatic precipitation of casein micelles without disrupting their internal structure. The reversibility of the interaction described opens the possibility of using this cationic polysaccharide to concentrate and resuspend casein micelles from whole or skim milk in the production of new fiber-enriched lactose-reduced calcium-caseinate dairy products.     

Use of surface plasmon resonance (SPR) to study the dissociation and polysaccharide binding of casein micelles and caseins

Tests were made to determine whether surface plasmon resonance (SPR) could be used as a technique to study the dissociation properties of bovine casein micelles or of sodium caseinate and the interactions between these protein particles and different polysaccharides. Surfaces of bound micelles or caseinate were made, and the changes in refractive index of these layers were used to define changes in the structures of the chemisorbed material. The technique appears to have some potential for studying details of the dissociation of casein micelles and of the binding of different polysaccharides to caseins.     

Study of the effect of soy proteins on the acid-induced gelation of casein micelles

The objective of this research was to understand whether addition of soy protein to milk protein affects the properties of acid-induced casein gels. Different samples were prepared by suspending casein micelles pellets in milk serum containing soy proteins or whey proteins as well as mixtures of the two proteins. Glucono-delta-lactone was added, and the changes in apparent size (in diluted systems) as well as the viscoelastic properties of the mixtures were measured. Size exclusion chromatography was also carried out to characterize the soluble phase of the various mixtures before and after heating. Soy protein affected the gelation of the mixtures; however, not to the same extent as whey proteins, which dominated formation of the network in soy-whey-casein systems. It was concluded that, up to a critical ratio of soy/whey proteins, soy proteins can be incorporated in the mix without a significant change in structure of the casein gels.     

Aggregation of casein micelles by interactions with chitosans: a study by Monte Carlo simulations

Recently, it was found that the addition of chitosan, a cationic polymer, to whole or skim milk produces the destabilization and coagulation of casein micelles which takes place without modifications in the milk pH or in the stability of most of the whey proteins. In the present work, Monte Carlo simulations are employed to show that the phase separation of casein micelles induced by chitosan can be explained by a depletion mechanism, where an effective attraction between the casein micelles is induced by the presence of chitosan molecules. This interaction is described on the basis of Vrij's model, where the depletion of polymer from the gap between neighboring casein micelles originates an effective attractive interaction that leads to a phase transition. This model, that considers volume restriction effects, accounts for several qualitative and even quantitative aspects of the experimental data for the coagulation of casein through chitosan addition.     

Effect of the anion citrate on the mineral composition of artificial casein micelles

The composition of artificial casein micelles (ACM) prepared at constant concentration of caseins, calcium (Ca), and phosphate (P(i)) in media with different citrate (Cit) concentrations was studied. The incorporation of the different mineral and protein components to the ACM was conditioned by the Cit concentration. In our working condition, the ACM size remained almost constant for Cit concentration ranging from 7 to 10 mM. This behavior could be indicating that the action of Cit essentially consists of a regulation of the Ca activity. The molar ratio at which Ca and P(i) were incorporated to the ACM varied for different Cit concentrations. At decreasing pH, the Ca/P(i) molar ratios for the remanent ions in the ACM were dependent on the Cit concentration. These observations could be related to a certain kind of competition between Cit, micellar calcium phosphate (MCP), and other groups able to bind Ca in the ACM.     

Interaction of environmental moisture with powdered green tea formulations: effect on catechin chemical stability

Green tea and tea catechins must be stable in finished products to deliver health benefits; however, they may be adversely affected by tea processing/storage conditions and the presence of other components. The objective of this study was to determine the effects of storage relative humidity (RH) and addition of other ingredients on catechin stability in simulated dry beverage mixtures. Samples of green tea powder alone and mixed with sucrose, citric acid, and/or ascorbic acid were prepared and stored in desiccators at 22 degrees C and 0-85% RH for up to 3 months. Epicatechin, epigallocatechin, epicatechin gallate, and epigallocatechin gallate were determined by high-performance liquid chromatography (HPLC). Formulation and the interaction of formulation and RH significantly promoted catechin degradation ( P < 0.0001). The chemical degradation of total and individual catechins in green tea powder formulations was significantly increased ( P < 0.0001) by exposure to increasing RH, and the degradation was exacerbated at > or = 58% RH by the presence of powdered citric acid and at > or = 75% RH by the presence of ascorbic acid. Catechins degraded the most in formulations containing both acids. Although catechin chemical stability was maintained at < or = 43% RH in all samples stored at 22 degrees C for 3 months, caking was observed in samples at these relative humidities. These results are the first to demonstrate that addition of other dry components to tea powders may affect catechin stability in finished dry blends and highlight the importance of considering the complex interplay between a multicomponent system and its environment for developing stable products.     

Effect of pH on the association of denatured whey proteins with casein micelles in heated reconstituted skim milk

Skim milk was adjusted to pH values between 6.5 and 6.7 and heated (80, 90, and 100 degrees C) for up to 60 min. Changes in casein micelle size, level of whey protein denaturation, and level of whey protein association with the micelles were monitored for each milk sample. Changes in casein micelle size were markedly affected by the pH at heating. At low pH (6.5-6.55), the casein micelle size increased markedly during the early stages of heating, and the size plateaued on prolonged heating. The maximum increase in size was approximately 30-35 nm. In contrast, at high pH (6.7), much smaller changes in size were observed on heating and the maximum increase in size was only approximately 10 nm. An intermediate behavior was observed at pH values between these two extremes. The rate of denaturation of the major whey proteins, alpha-lactalbumin and beta-lactoglobulin, was essentially unaffected by the pH at heating for the small pH changes involved in this study, and the changes in casein micelle size were poorly related to the level of whey protein denaturation. In contrast, the level of denatured whey proteins associating with the micelles was markedly dependent on the pH at heating, with high levels of association at pH 6.5-6.55 and low levels of association at pH 6.7. Changes in casein micelle size were related to the levels of denatured whey proteins that were associated with the casein micelles, although there was a small deviation from linearity at low levels of association (<15%). Further studies on reconstituted and fresh milk samples at smaller pH steps confirmed that the association of whey proteins with the casein micelles was markedly affected by the pH at heating. These results indicate that the changes in casein micelle size induced by the heat treatment of skim milk were a consequence of the whey proteins associating with the casein micelles and that the level of association was markedly influenced by small pH changes of the milk. It was not possible to determine whether the association itself influenced the casein micelle size or whether parallel reactions involving micellar aggregation caused the increase in micelle size as whey protein association progressed.     

Role of kappa-casein in the association of denatured whey proteins with casein micelles in heated reconstituted skim milk

Reconstituted skim milk at pH from 6.5 to 7.1 was unheated, preheated (68 degrees C/20 min), or heated at 90 degrees C for 20-30 min. On preheating, the size of the casein micelles decreased by about 5-20 nm, with a greater effect at higher pH. The casein micelle size of the heated milk at pH 6.5 increased by about 30 nm when compared to that of the unheated or preheated milk. As the pH was increased before heating, the particle size gradually decreased so that, at pH 7.1, the size was markedly smaller than that for the unheated milk and slightly smaller than that for the preheated milk. High levels (about 85%) of denatured whey protein associated with the casein micelles at pH 6.5, and this level decreased as the pH increased so that, at pH 7.1, low levels (about 15%) were associated with the micelles. Low levels of alphaS-casein and beta-casein were found in the serum regardless of the heat treatment or the pH of the milk. At pH 6.5, low levels (about 10%) of kappa-casein were also found in the milk serum. In the unheated milk, the level of serum kappa-casein increased slightly with increasing pH; in the heated samples, the level of serum kappa-casein increased markedly and linearly with increasing pH so that, at pH 7.1, about 70% of the kappa-casein was in the serum phase. The results of this study indicate that the pH dependence of the levels of serum phase kappa-casein may be responsible for the change in distribution of the whey proteins between the colloidal and serum phases. This is the first report to demonstrate significant levels of dissociation of kappa-casein from the micelles at pH between 6.5 and 6.7, although this dissociation phenomenon is well known on heating milk at high temperatures at pH above 6.7.     

Adsorption of xanthan polymer on allophane and effect on the changes in colloidal stability of allophane by heavy metal ions

The influence of divalent cations at concentrations of 10^-6.0 to 10^-4.0 M on the colloidal stability of partially deflocculated allophane by gum xanthan (GX) polysaccharide at pH 6.5 was investigated at two GX concentrations. Experiment in the presence of 10^-2.0 g L^-1 GX showed that the stability decreased by the addition of divalent cations and the effect of the decrease due to the cationic species was evident in a higher concentration range, i.e., 10^-5.0 to 10^-4.0 M. The order of the effect was Pb >Zn > Cd > Mg. Experiment in a 10^-4.5 g L^-1 GX solution revealed that (i) the stability increased by the addition of heavy metal cations at 10^-5.0 M (the order of the effect was Pb > Zn > Cd) and decreased at a concentration above 10^-4.5 M, (ii) whereas the stability decreased by the addition of Mg ion. The striking difference in the stability behavior due to the difference between the two GX-concentrations was attributed to the (i) degree of GX-adsorption (and hence negative charge from the carboxyl group in GX) onto allophane based on the electrophoretic mobility, (ii) complexation of heavy metal cations by organic ligand (carboxyl group) in GX which was adsorbed onto allophane, and (iii) surface complexation by heavy metal cations and hydroxyl groups on allophane. The results were discussed in relation to the characteristics of the particles of allophane, viz., (i) polymer-coated soft particles, and (ii) semi-soft particles on which the rigid (hard) surface of allophane substantially remained.     

Beta-lactoglobulin-dextran conjugates: effect of polysaccharide size on emulsion stability

A series of dextrans and beta-lactoglobulin were covalently conjugated and screened for their ability to stabilize oil-in-water emulsions. Dextrans with the molecular mass of 19.6 kDa, 87 kDa, 150 kDa, 500 kDa, and 2000 kDa were attached to beta-lactoglobulin via the Maillard reaction. The conjugates were then purified and evaluated as emulsifiers under neutral conditions. The ability to stabilize emulsions was determined by monitoring oil droplet size over time. Adsorption of the conjugates to the droplet surface was characterized by determining the protein surface load. The results show that increasing polysaccharide size increases emulsion stability up to 150 kDa before leveling off. Conversely, surface protein density remains constant until 150 kDa before decreasing with polysaccharide size. A model is presented to interpret the results.     

Effects of casein and fat content on water self-diffusion coefficients in casein systems: a pulsed field gradient nuclear magnetic resonance study

The water self-diffusion coefficients in casein matrixes were measured using a pulsed field gradient spin-echo nuclear magnetic resonance technique (PFG-SE NMR). The dependence of the water self-diffusion coefficient on the casein concentration and the aqueous phase composition is reported in both a rehydrated native phosphocaseinate dispersion and a concentrated casein retentate. A model has been proposed to explain the different behavior of the water self-diffusion coefficient in the two casein systems. This model demonstrates that the water self-diffusion cannot be simply explained by the water content only. So, taking into account the specific effect of each constituent of the aqueous dispersing phase, the water self-diffusion reduction induced by the casein micelle can be modeled. The effect of fat on the water self-diffusion coefficients was investigated. Anhydrous milk fat-reconstituted retentate samples were used in order to estimate the obstruction effect of fat globules in the modeling process. The dependence of the self-diffusion coefficient of water on the fat and casein content is reported. A general model included the effect of the aqueous phase composition, and the obstruction effects of casein micelles and fat globules were proposed. This model was validated for water self-diffusion coefficients in industrial fatty retentates.     

Studies on aggregate stability. 11. The effect of humic substances on the stability of re-formed soil aggregates

The effects of both physical addition and adsorption of humic substances on the aggregation of soils were studied. To investigate the effect of adsorbed humic materials onto soils it was necessary to develop an effective adsorption procedure. Mono-ionic soils (either sodium- or calcium-saturated) were used to ensure that adsorption had occurred and to enable the results to be properly interpreted. Physical addition of humic acid followed by incubation with glucose produced aggregates with low stability values. When humic acid was adsorbed onto the soil minerals and incubated, significant increases in aggregate stability were observed which persisted with time. The stability of the re-formed aggregates was even greater when the samples were incubated with glucose. This work has shown that humic substances were capable of stabilization of aggregates under difficult conditions where extracellular polysaccharides were ineffective, and that the stabilization had long term persistance. It is suggested that humic substances are particularly involved in the formation of micro-aggregates.     

Polyacrylamide polymer (PAM) effect on the propagation of the wetting front on a Jordanian soil under trickle irrigation system

Numerous studies have been conducted to examine the efficiency of PAM in controlling soil erosion under surface irrigation; however, there are fewer studies that have examined the effect of PAM on soil-water relations. This trickle irrigation laboratory study on a Jordanian clay loam soil addressed the effect of PAM on the spreading of the wetting front. PAM was added to the irrigation water under a trickle irrigation experiment where we irrigated the soil for 30 min. Four PAM concentrations (0.0, 5.0, 10.0, and 20.0 mg l^?1) were used to examine its effect on the wetting front propagation in the soil. We also examined the effect of PAM on the soil water content, where we irrigated for 45 min, using two concentrations of PAM, 0.0 and 20.0 mg l^?1. The soil water content was measured at depths of 0.0, 5.0, 10.0, 15.0, and 20.0 cm, 4 h, 24 h, and one week after completion of irrigation. PAM's effect on the soil water characteristic curve was studied using PAM concentrations of 0.0, 25, 50, and 100 mg l^?1. In this study the spread of the wetting front on the soil surface and in the vertical and horizontal directions inside the soil profile increased when PAM was added to the irrigation water. As the PAM concentration increased the spreading of the wetting front increased. The average wetting front spreading increased in the vertical direction from 7.8 cm when PAM was not used to 8.3, 9.6, and 12.1 cm, for PAM concentrations of 5.0, 10.0, and 20.0 mg l^?1, respectively. At the same time the average increase in the horizontal direction was from 18.4 to 18.5, 18.6, and 21.7 cm for PAM concentrations of 0.0, 5.0, 10.0, and 20.0 mg l^?1, respectively. The second experiment showed that at a soil depth of 20 cm, volumetric soil water content (θv) was 15.2% when the irrigation water contained 20.0 mg l^?1 of PAM compared to 13.9% when no PAM was added to the irrigation water. PAM did not have a significant effect on soil moisture characteristic curve.     

Electrochemical investigation of the effect of pH and solvent on amitraz stability

The widespread use of the pesticide amitraz for pest control of crops, livestock and honeybees has warranted several studies aimed at understanding the degradation of this compound during storage and use. In particular the degradation of amitraz and the nature of the toxicologically significant intermediates formed owing to pH and solvent type has been examined. In this study we report on the use of electrochemical methods to monitor amitraz degradation and to identify the major intermediates formed. While this study examines the use of rapid voltammetric methods for such analyses, it also resolves earlier studies showing the rapid degradation of amitraz to 2,4-dimethylaniline without formation of intermediates first, and also suggests that the degradation of amitraz to 2,4-dimethylphenylformamide and to 2,4-dimethylaniline is more rapid than previously observed at pH above 3. These studies also showed that amitraz degrades to dimethylphenylformamide in ethanol and methanol, and is stable in both acetonitrile and dimethylsulphoxide.     

Interaction of flavonoids with bovine serum albumin: a fluorescence quenching study

The interaction between four flavonoids (catechin, epicatechin, rutin, and quercetin) and bovine serum albumin (BSA) was investigated using tryptophan fluorescence quenching. Quenching constants were determined using the Stern-Volmer equation to provide a measure of the binding affinity between the flavonoids and BSA. The binding affinity was strongest for quercetin and ranked in the order quercetin > rutin > epicatechin = catechin. The pH in the range of 5-7.4 does not affect significantly (p < 0.05) the association of rutin, epicatechin, and catechin with BSA, but quercetin exhibited a stronger affinity at pH 7.4 than at lower pH (p < 0.05). Quercetin has a total quenching effect on BSA tryptophan fluorescence at a molar ratio of 10:1 and rutin at approximately 25:1. However, epicatechin and catechin did not fully quench tryptophan fluorescence over the concentration range studied. Furthermore, the data suggested that the association between flavonoids and BSA did not change molecular conformation of BSA and that hydrogen bonding, ionic, and hydrophobic interaction are equally important driving forces for protein-flavonoid association.     

The effect of optimization of silicon nutrition on the stability of barley DNA

Numerous studies on the use of silicon fertilizers indicate their positive influence on the resistance of plants to biogenic and abiogenic stress; nevertheless, the mechanisms of action of silicon on the plants have been little studied. Finely divided amorphous silica was used as a reference silicon fertilizer in microfield experiments that were carried out in the north of the Moscow region with a culture of barley. The strengthening of DNA molecules was shown to occur at doses of SiO₂ of 50, 500, and 1000 kg/ha, which provided higher concentrations of monosilicic acid in the soil than other doses. Therefore, the introduction of silicon improves the viability of the plants at the genome level and enhances the natural resistance of plants in agricultural lands.     

Calcium chloride washing of calcareous sediment from a freshwater canal: effect on the removal of potentially toxic elements and water aggregate stability

Journal of Soils and Sediments - Canal dredging and shaping produce considerable amounts of sediments whose reuse on- and off-site depends on their pollution level. This study explores the...