Disruption and reassociation of casein micelles under high pressure: influence of milk serum composition and casein micelle concentration

In this study, factors influencing the disruption and aggregation of casein micelles during high-pressure (HP) treatment at 250 MPa for 40 min were studied in situ in serum protein-free casein micelle suspensions. In control milk, light transmission increased with treatment time for approximately 15 min, after which a progressive partial reversal of the HP-induced increase in light transmission occurred, indicating initial HP-induced disruption of casein micelles, followed by reformation of casein aggregates from micellar fragments. The extent of HP-induced micellar disruption was negatively correlated with the concentration of casein micelles, milk pH, and levels of added ethanol, calcium chloride, or sodium chloride and positively correlated with the level of added sodium phosphate. The reformation of casein aggregates during prolonged HP treatment did not occur when HP-induced disruption of casein micelles was limited (<60%) or very extensive (>95%) and was promoted by a low initial milk pH or added sodium phosphate, sodium chloride, or ethanol. On the basis of these findings, a mechanism for HP-induced disruption of casein micelles and subsequent aggregation of micellar fragments is proposed, in which the main element appears to be HP-induced solubilization of micellar calcium phosphate.     

Ethanol-dependent heat-induced dissociation of casein micelles

The dissociation of casein micelles when heated to approximately 65 degrees C in the presence of ethanol [1:1 mixture (v/v) of milk and 65% (w/w) aqueous ethanol] was investigated using L* values and transmission measurements. Mixtures of milk and ethanol became transparent on heating, which suggests dissociation of casein micelles. Results of experiments using confocal laser scanning microscopy, light scattering (static and dynamic), and dialysis to examine the changes of milk during heating in the presence of ethanol supported the assertion that such treatments result in dissociation of casein micelles, as did studies of model beta-casein micellar systems.     

Mechanism for the ethanol-dependent heat-induced dissociation of casein micelles

An explanation as to how casein micelles dissociate when heated in the presence of ethanol is presented. Dissociation of casein micelles in milk-ethanol mixtures was studied using (1)H NMR, and the effects of addition of CaCl(2), NaCl, or EDTA or alteration of milk pH on this dissociation were studied. It is proposed that at low temperatures, ethanol reduces the solvent quality of milk serum, but above a critical temperature (approximately 30 degrees C in a 35% ethanol solution), ethanol enhances solvent quality and dissociates the casein micelles. Ethanol reduced protein hydrophobicity and increased the pK(a) value of phosphoserine, effects that are likely to be significant in the dissociating effect of ethanol at elevated temperatures.     

Aggregation of rennet-altered casein micelles at low temperatures

The rennet-induced coagulation of bovine milk at 10 degrees C was investigated. The rate of change of absorbance at 600 nm was higher in milk renneted at 30 degrees C than that at 10 degrees C. The amount of casein sedimented on centrifuging skim milk at 5000g for 1 h at 10 degrees C increased with time after renneting. The viscosity of milk at 10 degrees C at low shear rates did not change significantly until 10 h after rennet addition, but it increased markedly after 20 h. Smaller particles in milk at 10 degrees C disappeared slowly over 36 h after rennet addition and aggregated into larger particles. These results suggested that casein micelles in milk aggregate at low temperatures. Reasons for the slow aggregation of milk renneted at 10 degrees C were investigated by inhibiting chymosin activity by pepstatin A. It is likely that beta-casein, or its hydrolysis, plays a role in aggregation of rennet-altered casein micelles at low temperatures.     

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.     

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.     

Solvent-mediated disruption of bovine casein micelles at alkaline pH

The disruption of casein micelles at alkaline pH was investigated using turbidity measurements. The rate and extent of disruption of casein micelles at alkaline pH (8.0-11.0) increased with pH. Furthermore, the extent of alkaline disruption increased with increasing temperature (5-40 degrees C). Preheating milk for 10 min at 90 degrees C did not influence the extent of alkaline disruption of casein micelles, suggesting that whey proteins do not influence the alkaline disruption process. Levels of ionic calcium and serum calcium and phosphate decreased in a logarithmic fashion with increasing pH, indicating precipitation of calcium phosphate onto the casein micelles. A mechanism for alkaline disruption of casein micelles is proposed, in which increasing the milk pH improves the solvent quality for the caseins, thereby leading to the disruption of casein micelles into their constituent nanoclusters; increases in the net-negative charge on the caseins on increasing pH may contribute to micellar dissociation.     

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.     

Effect of turnip mosaic virus infection on the mineral composition of rutabaga

Abstract

The effect of Turnip Mosaic Virus (TuMV) infection on the elemental composition of rutabaga (Brassica napus ssp. rapifera (Metzg) Sinsk) at maturity was examined in the greenhouse. TuMV infection markedly reduced dry weight and dry matter percentage of leaves‐plus‐petioles but did not affect storage root development. Virus‐infected plants exhibited higher N, P, Mg and Zn, but lower K levels in leaf tissues. Nitrogen, Ca, Mg and Mn accumulated in storage root tissues as a result of TuMV infection.     

Effect of spectral composition of artificial light on the attraction of moths

During the last decades, artificial night lighting has increased globally, which largely affected many plant and animal species. So far, current research highlights the importance of artificial light with smaller wavelengths in attracting moths, yet the effect of the spectral composition of artificial light on species richness and abundance of moths has not been studied systematically. Therefore, we tested the hypotheses that (1) higher species richness and higher abundances of moths are attracted to artificial light with smaller wavelengths than to light with larger wavelengths, and (2) this attraction is correlated with morphological characteristics of moths, especially their eye size. We indeed found higher species richness and abundances of moths in traps with lamps that emit light with smaller wavelengths. These lamps attracted moths with on average larger body mass, larger wing dimensions and larger eyes. Cascading effects on biodiversity and ecosystem functioning, e.g. pollination, can be expected when larger moth species are attracted to these lights. Predatory species with a diet of mainly larger moth species and plant species pollinated by larger moth species might then decline. Moreover, our results indicate a size-bias in trapping moths, resulting in an overrepresentation of larger moth species in lamps with small wavelengths. Our study indicates the potential use of lamps with larger wavelengths to effectively reduce the negative effect of light pollution on moth population dynamics and communities where moths play an important role.     

Effect of chromium VI on mineral element composition of bush beans

Bush beans (Phaseolus vulgaris L. cv Contender) were grown on perlite with nutrient solution and 0, 1, 2.5 and 5 ppm levels of Na₂CrO₄ Significant decrease of top growth and chlorosis in trifoliated leaves were observed for 2.5 and 5 ppm Cr, with Cr concentrations (μg/g) in tops:≥ 12.1, in roots:≥ 509.9. Cr decreased K, Na, Mg and Fe concentrations, and increased P and Mn concentrations in roots. In tops decreased N, K, Na and Fe concentrations and increased Mn and Ca concentrations were observed, Translocation of P, Zn, Cu and Fe was inhibited; Ca and Mn translocation was generally enhanced. P/Fe ratio was increased up to 60% in chlorotic plants, indicating a shift from Fe²⁺ to Fe³⁺.     

Effect of light and water stress on mineral element composition of plants 1

Many factors affect the composition of mineral elements in plants. The effects of many of these factors, such as element concentration, temperature, pH, plant part, plant age, metabolic inhibitors, element interactions, fertilizer, and type of soil, have been studied extensively by many scientists who are concerned with plant mineral nutrition. However, relatively few studies have focused on the effects of light and water stress. The objective of this review was to consider some of the effects that light and water stress have on the mineral element composition of plants.     

Effect of anion composition of simulated acid rain on nutrient behavior in reclaimed saline soils

Abstract

Field experiments with the “Taikichi” taro cultivar were conducted in volcanic ash soil of Kagoshima Prefecture, Japan, in order to determine the effects of potassium applied with fertilizers and manures on the growth and yield of taro. The experimental design was accord ing to the L27 (3¹³) orthogonal factorial experiment, the three factors selected being the manures, nitrogen fertilizers and amount of potassium application.

“Metsubure” corm formation was infrequent in hog manured plots without potassium application, but was significantly increased by the application of potassium fertilizer. Furthermore, the occurrence of “Metsubure” corms was significantly lower in fields prepared with hog manure than in those prepared with plant residue and cattle manure.

Chemical analysis of the taro plants and manures revealed that the plants absorbed much more calcium when planted in hog manure than in other manures. The CaO/K₂O ratio was also higher. The main reason for this was assumed to be the lower content of potassium in the hog manure (0.37% K₂O on a dry basis).

The analytic results suggest that potassium disturbs the calcium uptake ability of the taro plant causing “Metsubure” corm formation.

The authors also attempted to determine the effects of several kinds of nitrogen fertilizers on “Metsubure” corm formation, but no clear results were obtained in this experiment.     

Effect of salinity on stomatal resistance,proline, and mineral composition of pepper

Pepper (Capsicum annuum L.) plants grown in pots were irrigated with the nutrient solutions containing 50, 75, and 100 mM NaCl or a control solution. Salinity markedly decreased plant growth. Increasing salinity levels increased stomatal resistance and sodium (Na), chloride (Cl), proline contents of the plants. Potassium (K), total‐nitrogen (N), and chlorophyll content of the plants were decreased under high salinity conditions.     

Effect of rate and source of nitrogen fertilizers on mineral composition of d'Anjou pears

Abstract

Thirteen‐year‐old ‘d'Anjou’ pear trees, Pyrus communis L., were fertilized with 3 rates of ammonium nitrate or 2 rates of calcium nitrate in late autumn from 1978 to 1980. In 1981, mid‐terminal and fruiting‐spur leaves and fruit peel and flesh were sampled for mineral analysis of N, P, K, Ca, Mg, B, Cu, Fe, Mn, and Zn. The level of N increased in the above tissues as rate of N fertilizer was increased. In most cases, the levels of N, P, K, Mg, Cu, and Mn in the fruiting spur leaves and fruit were greater in the ammonium nitrate‐treated trees than with calcium nitrate fertilizer. Calcium was higher in the fruit peel and flesh of trees treated with the low rate of calcium nitrate than with the highest rate of ammonium nitrate fertilizer while Ca in the fruiting spur leaves was lower in the calcium nitrate‐fertilized trees.     

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 salinity on growth,mineral composition,and water relations of grafted tomato plants

A greenhouse experiment was designed in order to evaluate growth, water relations, and nutrient concentrations of grafted and ungrafted tomato plants grown under varying levels of salinity (0, 30, or 60 mM NaCl). Two cultivars, ‘Fanny’ and ‘Goldmar’, were grafted onto AR‐9704, using the cleft‐grafting method. Growth of grafted ‘Fanny’ plants was higher than that of ungrafted plants. Growth of ‘Goldmar’ plants was not affected by salinity treatments or grafting, but it was slower than for ‘Fanny’. Leaf turgor showed no significant differences between grafted and ungrafted plants or between salinity levels. The stomatal conductance (G_s) was higher for grafted than for ungrafted plants, and salinity decreased it significantly and progressively in both grafted and ungrafted plants and in both varieties. The concentrations of Na⁺ and Cl^– were significantly higher in ungrafted than in grafted ‘Fanny’ plants. ‘Fanny’ was more tolerant when grafted, probably due to reduced accumulation of Na⁺ and/or Cl^– in the shoot.     

Interaction of a cationic acrylate polymer with caseins: biphasic effect of Eudragit E100 on the stability of casein micelles

When whole or skim milk was incubated with the cationic acrylate polymer Eudragit E100, a biphasic effect on the stability of casein micelles was observed. A precipitation phase was observed at low polymer/casein ratios. Strikingly, a solubilization phase of the aggregates was observed when the ratios of polymer/casein were increased. Purified alpha(s)-, beta-, and kappa-caseins or dephosphorylated caseins were equally precipitated and resolubilized by the cationic polymer, indicating no special selectivity for a particular protein or phosphate residue for these events. An increase in the size of the aggregates as the optimum precipitating amount of Eudragit E100 was reached suggests a crossbridging of the micelles by the polymer. The inhibition of the precipitation phase by high ionic strength indicates that electrostatic interactions play a critical role in complex formation. Furthermore, a dramatic reduction in size of the protein colloidal particles upon solubilization of the aggregates was observed by dynamic light scattering, indicating a dissociation of the micellar structure. Taken together, the results indicate that at low concentration Eudragit E100 may act as a precipitant of casein micelles, mainly by ionic interaction and at high concentration as an amphipathic agent, solubilizing casein micelles with a disruption of their internal structure.     

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.