Seasonal variations in composition,properties, and heat-induced changes in bovine milk in a seasonal calving system

We determined seasonal variations in the composition and characteristics of bovine milk, as well as heat-induced changes in the physicochemical properties of the milk, in a typical seasonal-calving New Zealand herd over 2 full milking seasons. Fat, protein, and lactose contents varied consistently during the year in patterns similar to those of the lactation cycle. Seasonality also had significant effects on milk calcium, ionic calcium, fat globule size, buffering capacity, and ethanol stability, but not on casein micelle size. The ratio of casein to total protein did not vary significantly over the season, but late-season milk had the highest content of glycosylated κ-casein (G-κ-CN) and the lowest content of α-lactalbumin in both years. We observed significant between-year effects on protein, total calcium, ionic calcium, pH, and casein:total protein ratio, which might have resulted from different somatic cell counts in the 2 years. Compared with heating at 90°C for 6 min, UHT treatment (140°C for 5 s) induced greater dissociation of κ-casein, a similar extent of whey protein denaturation, a lower extent of whey protein–casein micelle association, and a larger increase in casein micelle size. Indeed, UHT treatment might have triggered significant dissociation of G-κ-CN, resulting in aggregation among the casein micelles and increased apparent mean casein micelle diameter. Seasonality had significant effects on the partitioning of G-κ-CN between the micelle and the serum phase, the extent of whey protein–casein micelle association under both heating conditions, and the casein micelle size of the UHT milk.     

IDENTIFICATION OF HEAT SENSITIVE PROTEINS DURING ROASTING OF PEANUT SEED

Peanut (Arachis hypogaea L) seed were heated at 125C and 150C between 0 to 24 min and the seed proteins were extracted with 0.1 M sodium phosphate buffer, pH 2.5. The soluble and insoluble proteins were analyzed to determine changes in protein and polypeptide composition during heating. Acid soluble proteins were more susceptible to thermal breakdown than acid insoluble proteins. Within the acid soluble fraction, some proteins and polypeptides were more readily degraded than the others. In the acid insoluble fraction, except for the loss of a 70,000 Dalton polypeptide no other major changes were observed. Changes in protein composition were more rapid at 150C heating than at 125C heating. These data suggest that the proteins of the acid soluble fraction are highly sensitive to heating and thus may have a role in flavor volatile production during roasting of the peanut.     

Effects of depleting ionic strength on 31P nuclear magnetic resonance spectra of micellar casein during membrane separation and diafiltration of skim milk

Membrane separation processes used in the concentration and isolation of micellar casein-based milk proteins from skim milk rely on extensive permeation of its soluble serum constituents, especially lactose and minerals. Whereas extensive literature exists on how these processes influence the gross composition of milk proteins, we have little understanding of the effects of such ionic depletion on the core structural unit of micellar casein [i.e., the casein phosphate nanocluster (CPN)]. The ³¹P nuclear magnetic resonance (NMR) is an analytical technique that is capable of identifying soluble and organic forms of phosphate in milk. Thus, our objective was to investigate changes to the ³¹P NMR spectra of skim milk during microfiltration (MF) and diafiltration (DF) by tracking movements in different species of phosphate. In particular, we examined the peak at 1.11 ppm corresponding to inorganic phosphate in the serum, as well as the low-intensity broad signal between 1.5 and 3.0 ppm attributed to casein-associated phosphate in the retentate. The MF concentration and DF using water caused a shift in the relevant ³¹P NMR peak that could be minimized if orthophosphate was added to the DF water. However, this did not resolve the simultaneous change in retentate pH and increased solubilization of micellar casein protein. The addition of calcium in combination with orthophosphate prevented micellar casein solubilization and simultaneously contributed to preservation of the CPN structure, except for overcorrection of retentate pH in the acidic direction. A more complex DF solution, involving a combination of phosphate, calcium, and citrate, succeeded in both CPN and micellar casein structure preservation while maintaining retentate pH in the region of the original milk pH. The combination of ³¹P NMR as an analytical technique and experimental probe during MF/DF processes provided useful insights into changes occurring to CPN while retaining the micellar state of casein.     

Structural analysis of the environment of calcium ions in crystalline and amorphous calcium phosphates by X-ray absorption spectroscopy and a hypothesis concerning the biological function of the casein micelle

Determination of the structural environment around calcium ions, using X-ray absorption spectroscopy, in crystalline calcium phosphates is described. The spherically averaged short-range order in an amorphous calcium phosphate can be quantitatively determined using, as a model, the environment of calcium ions in the crystalline calcium phosphate which is closest to it in chemical composition. Structural studies, together with other evidence, suggest that the colloidal calcium phosphate in milk comprises small clusters of Ca²⁺ and HPO₄²⁻ ions in a fairly disordered state. The ion clusters are distributed throughout the micelle and are probably linked through phosphoseryl residues, to the Ca²⁺-sensitive caseins. Calculations indicate that probably about five polypeptide chains interact with a typical ion cluster.The small size of the ion clusters, their amorphous nature and the relative conformational flexibility of the casein proteins suggests a possible biological function for caseins and the casein micelle in the control of calcium phosphate precipitation. During calcium transport by the secretory cells of the mammary epithelium, calcium phosphate is precipitated in the Golgi vesicles. It is suggested that the caseins trap the growth of the precipitate at the nucleation stage, effectively preventing a potentially disastrous first-order phase transition.     

Changes in the calcium cluster distribution of ultrafiltered and diafiltered fresh skim milk as observed by Small Angle Neutron Scattering

The effect of ultrafiltration and diafiltration on the distribution of the calcium phosphate clusters of the casein micelle was investigated using Small Angle Neutron Scattering (SANS). In the case of ultrafiltration, fresh skim milk was subjected to concentration using membrane filtration up to 5× its original volume, the retentate was rediluted with its corresponding serum and subsequently dialyzed against reconstituted milk powder dispersed in D2O/H2O (UF 5×(D)). In the case of diafiltered samples, the samples were concentrated adding water (diafiltration) at two different levels (DF 2·5× or DF 5×) and then redispersed with D2O/H2O. In the DF 5× case, the serum components were diluted to less than 1% of their original concentration. For analysis, all samples had the same volume fraction of dispersed casein micelles (φ=0·1), which is that of the control, unprocessed skim milk. A peak in the SANS data was observed in fresh skim milk at a scattering vector, qo, of 0·034 ?-1 (directly proportional to the reciprocal characteristic length), in agreement with previous literature results. Neutron data on the ultrafiltered, UF 5×(D) and diafiltered, DF 2·5× and DF 5× milk samples showed a progressive decrease in the intensity of the peak but invariance in qo. These results, combined with the determination of soluble and insoluble calcium in the samples, suggest a progressive and irreversible removal of calcium from within the micelle during membrane filtration of milk. Using SANS it was possible to clearly show changes in the micellar calcium clusters that may not otherwise be fully determined by only measuring the amount of total and insoluble calcium in milk.     

Association of denatured whey proteins with casein micelles in heated reconstituted skim milk and its effect on casein micelle size

When skim milk at pH 6.55 was heated (75 to 100 degrees C for up to 60 min), the casein micelle size, as monitored by photon correlation spectroscopy, was found to increase during the initial stages of heating and tended to plateau on prolonged heating. At any particular temperature, the casein micelle size increased with longer holding times, and, at any particular holding time, the casein micelle size increased with increasing temperature. The maximum increase in casein micelle size was about 30-35 nm. The changes in casein micelle size were poorly correlated with the level of whey protein denaturation. However, the changes in casein micelle size were highly correlated with the levels of denatured whey proteins that were associated with the casein micelles. The rate of association of the denatured whey proteins with the casein micelles was considerably slower than the rate of denaturation of the whey proteins. Removal of the whey proteins from the skim milk resulted in only small changes in casein micelle size during heating. Re-addition of beta-lactoglobulin to the whey-protein-depleted milk caused the casein micelle size to increase markedly on heat treatment. The changes in casein micelle size induced by the heat treatment of skim milk may be a consequence of the whey proteins associating with the casein micelles. However, these associated whey proteins would need to occlude a large amount of serum to account for the particle size changes. Separate experiments showed that the viscosity changes of heated milk and the estimated volume fraction changes were consistent with the particle size changes observed. Further studies are needed to determine whether the changes in size are due to the specific association of whey proteins with the micelles or whether a low level of aggregation of the casein micelles accompanies this association behaviour. Preliminary studies indicated lower levels of denatured whey proteins associated with the casein micelles and smaller changes in casein micelle size occurred as the pH of the milk was increased from pH 6.5 to pH 6.7.     

Structural changes induced by high-pressure processing in micellar casein and milk protein concentrates

Reconstituted micellar casein concentrates and milk protein concentrates of 2.5 and 10% (wt/vol) protein concentration were subjected to high-pressure processing at pressures from 150 to 450 MPa, for 15 min, at ambient temperature. The structural changes induced in milk proteins by high-pressure processing were investigated using a range of physical, physicochemical, and chemical methods, including dynamic light scattering, rheology, mid-infrared spectroscopy, scanning electron microscopy, proteomics, and soluble mineral analyses. The experimental data clearly indicate pressure-induced changes of casein micelles, as well as denaturation of serum proteins. Calcium-binding α_S1- and α_S2-casein levels increased in the soluble phase after all pressure treatments. Pressurization up to 350 MPa also increased levels of soluble calcium and phosphorus, in all samples and concentrations, whereas treatment at 450 MPa reduced the levels of soluble Ca and P. Experimental data suggest dissociation of calcium phosphate and subsequent casein micelle destabilization as a result of pressure treatment. Treatment of 10% micellar casein concentrate and 10% milk protein concentrate samples at 450 MPa resulted in weak, physical gels, which featured aggregates of uniformly distributed, casein substructures of 15 to 20 nm in diameter. Serum proteins were significantly denatured by pressures above 250 MPa. These results provide information on pressure-induced changes in high-concentration protein systems, and may inform the development on new milk protein-based foods with novel textures and potentially high nutritional quality, of particular interest being the soft gel structures formed at high pressure levels.     

Hydration of casein micelles and caseinates: Implications for casein micelle structure

By studying the hydration of casein micelles using a variety of techniques, a distinction could be made between water that appeared bound by the protein (∼0.5 g g⁻¹ protein), water associated with the κ-casein brush (∼1.0 g g⁻¹ protein) and water entrapped in the casein micelles (∼1.8 g g⁻¹ protein), yielding a total micellar hydration of ∼3.3 g g⁻¹ protein, in line with casein micelle voluminosity derived from intrinsic viscosity measurements. For caseinate particles, however, the main contributor to intrinsic viscosity was not protein hydration but the non-spherical particle shape. These non-spherical particles in caseinate are likely to be naturally present as primary casein particles (PCP) in casein micelles. PCP could be used to build casein micelles by controlled introduction of micellar salts. Based on the findings of this study, casein micelles could be described as a porous network of non-spherical PCP linked by calcium phosphate nanoclusters.     

Effect of sodium citrate on structure-function relationships of Cheddar cheese

The objective of this study was to determine the effect of sodium citrate on the structure and functionality of Cheddar cheese. The hypothesis was that citrate (sodium citrate) injection would affect cheese properties mainly through its effect on bound calcium (calculated as the difference between total calcium and the water-soluble calcium content of a cheese extract). A 9-kg block of Cheddar cheese was made, vacuum-packaged, and then stored for 2 wk at 4 degrees C. After storage, the cheese was cut into 0.5- to 0.6-kg blocks that were vacuum-packaged and stored for 1 wk at 4 degrees C prior to injection. Cheese blocks were then high-pressure injected with a buffer solution (pH 5.27) containing 40% (wt/ wt) citric acid trisodium dihydrate and 6.25% (wt/wt) anhydrous citric acid, from zero (control) to five times (successive injections performed 24 h apart). Increased citric acid content of cheese from 0.22 (uninjected) to 1.39% (after five injections) caused phosphate solubilization. Thus, the calculated bound phosphate content of cheese decreased from 0.54 to 0.45 mmol/g of protein. However, unexpectedly, the soluble calcium content decreased from 0.34 (control) to 0.28 mmol/g of protein (after five injections), whereas the bound calcium content remained unchanged (0.42 mmol/g of protein). The decrease in soluble calcium probably resulted from the formation and concentration of crystals in the cheese surface, which was not included in samples for analysis, and from the expulsion of serum from within the cheese. Higher concentration of solutes in the water phase of cheese would increase the volume of serum, but the cheese had limited holding capacity and serum was expelled. Citrate injection increased the sodium content of cheese from 0.63 to 0.93%, but it had no effect on cheese pH (5.2). After five injections, the protein matrix expanded, occupying an increased area of cheese matrix (83 vs. 78%). Even though citrate injection had no effect on bound calcium, and thus the rate and extent of cheese flow were unaffected, increased phosphate solubilization, and possibly decreased ionic calcium content, resulted in expansion of the protein matrix and increased cheese hardness.     

Fluorescent labeling study of plasminogen concentration and location in simulated bovine milk systems

A fluorescent labeling method was developed to study plasminogen (PG) concentration and location in simulated bovine milk. Activity and stability of PG labeled with Alexa Fluor 594 (PG-594) were comparable to those of native PG. The fluorescent signal of PG-594 exhibited pH, temperature, and storage stability, and remained stable throughout typical sample treatments (stirring, heating, and ultracentrifugation). These characteristics indicate broad applicability of the fluorescent labeling technique for milk protease characterization. In an example application, PG-594 was added to simulated milk samples to study effects of heat and β-lactoglobulin (β-LG) on the distribution of PG. Before heating, about one-third of the PG-594 remained soluble in the whey fraction (supernatant) whereas the rest became associated with the casein micelle. Addition of β-LG to the system slightly shifted PG-594 distribution toward the whey fraction. Heat-induced PG-594 binding to micelles in whey-protein-free systems was evidenced by a decrease of PG-594 from 31 to 15% in the whey fraction accompanied by an increase of PG-594 from 69 to 85% in casein micelle fractions. When β-LG was present during heating, more than 95% of PG-594 became associated with the micelle. A comparison with the distribution pattern of PG-derived activities revealed that heat-induced PG binding to micelles accompanies heat-induced PG inactivation in the micelle fraction. Incubation of the casein micelles with the reducing agent β-mercaptoethanol revealed that disulfide bonds formed between PG and casein or between PG and casein-bound β-LG are the mechanisms for heat-induced PG binding to casein micelles. Western blotting and zymography results correlated well with fluorescent labeling studies and activity studies, respectively. Theoretically important findings are: 1) when heated, serum PG is capable of covalently binding to micellar casein or complexing with β-LG in whey and then coadhering to micelles, and 2) PG that associated with micellar casein through lysine binding sites before heating is capable of developing heat-induced disulfide bonds with casein. The overall results are PG covalently binding to micelles and inactivation thereafter. Our results suggest that, instead of thermal denaturation through irreversible unfolding, covalent bond formation between PG and other milk proteins is the mechanism of PG inhibition during thermal processing.     

Experimental evidence for previously unclassified calcium phosphate structures in the casein micelle

¹H-³¹P Cross-polarization magic angle spinning (CP-MAS) measurements of 40-d-old Mozzarella cheese and 20 mM EDTA-treated casein micelles revealed that each sample had immobile phosphorus with the same spectral pattern, which did not match that of native casein micelles. To identify the immobile phosphorus bodies, ¹H-³¹P CP-MAS spectra and cross-polarization kinetics measurements were undertaken on native casein micelles, EDTA-chelated casein micelles, and reference samples of β-casein and hydroxyapatite. The results showed that the immobile phosphorus bodies in the mature Mozzarella cheese had the following characteristics: they are immobile phosphoserine residues (not colloidal calcium phosphate); they are not the product of phosphoserine to colloidal calcium phosphate bridging; the phosphate is complexed to calcium; their rigidity is localized to a phosphorus site; their rigidity and bond coupling is unaffected by protein hydration; and the immobile bodies share a narrow range of bond orientations. Combining these observations, the best explanation of the immobile phosphorus bodies is that bonding structures of phosphorus-containing groups and calcium exist within the casein micelle that are not yet clearly classified in the literature. The best candidate is a calcium-bridged phosphoserine-to-phosphoserine linkage, either intra- or inter-protein.     

Distribution of Ca,P and Mg and casein micelle mineralisation in donkey milk from the second to ninth month of lactation

Ca, P and Mg content and distribution between soluble and colloidal phases of donkey milk (DM) were investigated in 62 individual milk samples collected from 9 Ragusano donkeys followed from the second to the ninth month of lactation. Ca (69%) and P (64%) were mainly associated with casein, while Mg was largely found in the soluble form (69%). Only 25% of the colloidal P was present as phosphorylated residues of caseins (casein-P). The colloidal contents of Ca, casein-P, inorganic P (as a constituent of the colloidal calcium phosphate; CCP) and Mg were 1.84, 1.30, 0.45 and 0.15 mmol g casein⁻¹, respectively, revealing a high level of mineralisation of DM casein micelle. The colloidal Ca to colloidal inorganic-P ratio was 1.51, suggesting that most colloidal Ca was in the form of CCP. All parameters were affected by the period of lactation, except casein level in milk and its level of phosphorylation.     

Effects of mineral salts and calcium chelating agents on the gelation of renneted skim milk

The effects of adding CaCl2, orthophosphate, citrate, EDTA, or a mixture of these, to reconstituted skim milk (90 g of solids/kg solution) on the gelation of renneted milk were mediated by changes in Ca2+ activity and the casein micelle. At pH 6.65, the addition of citrate or EDTA, which removed more than 33% of the original colloidal calcium phosphate with the accompanying release of 20% casein from the micelle, completely inhibited gelation. Reformation of the depleted colloidal calcium phosphate and casein in the micelle, by the addition of CaCl2, removed this inhibition. When the minimum requirements for colloidal calcium phosphate and casein in the micelle were met, the coagulation time decreased with increasing Ca2+ activity, leveling off at high Ca2+ activity. The storage modulus of renneted gels, measured at 3 h, increased with increasing colloidal calcium phosphate content of micelles up to a level at which it was approximately 130% of the original colloidal calcium phosphate in the micelles. Further increases in colloidal calcium phosphate by the addition of CaCl2, orthophosphate, or mixtures of these, which did not change the proportion of casein in the micelle, decreased the storage modulus. The gelation of the renneted milk was influenced by Ca2+ activity, the amounts of colloidal calcium phosphate, and casein within the micelle, with the effects of colloidal calcium phosphate and casein within the micelle clearly dominating the storage modulus. These results are consistent with the model of Horne (Int. Dairy J. 8:171-177, 1998) which postulates that, following cleavage of the stabilizing K-casein hairs by rennet, the properties of the rennet gel are determined by the balance between the electrostatic and hydrophobic forces between casein micelles.     

Gelation Mechanism of Soybean 11S Globulin: Formation of Soluble Aggregates as Transient Intermediates

Thermal association-dissociation behavior of soybean 11S globulin was investigated by sucrose density gradient centrifugation and polyacrylamide gel electrophoresis. Soluble aggregates with a molecular weight of 8 × 10⁶ were formed when 0.5% and 5% protein solutions were heated for 1 min at 100°C. At the lower protein concentration, subsequent heating caused disappearance of the soluble aggregate followed by complete dissociation into acidic and basic subunits. At the higher concentration, however, subsequent heating caused formation of highly polymerized aggregates, and gel was formed after 5 min of heating. The soluble aggregates appear to be transient intermediates in the course of gel formation of 11S globulin.     

The liquid-state 31P-nuclear magnetic resonance study on microfiltrated milk

To gain further insight into diversiform phosphorus in bovine milk, we separated skim milk into casein micelle and serum fractions by microfiltration and subjected them to liquid-state 31P-nuclear magnetic resonance (NMR) spectroscopy. As previously reported, the skim milk spectrum showed a broad and indistinct peak from phosphoserine residue (SerP) of casein. In the casein micelle spectrum, however, the SerP peak was more clearly observed with a phosphate peak that may be from micellar calcium phosphate (MCP). The serum spectrum was the same as skim milk spectrum, except for SerP peak. Furthermore, two types of casein micelle fractions, with 0.90 and 1.04 of [beta-casein + kappa-casein]/[alpha(s1)-casein + alpha(s2)-casein] ratios were generated by different temperature microfiltrations, occurring because beta-casein is released from the micelle at a low temperature. The shape of SerP peaks changed dramatically in both the casein micelle spectra, when the temperature dropped from 35 to 5 degrees C. Deconvolution analysis indicated that each SerP peak comprised the same set of four peaks. Half-width and composition discriminated between the two types of casein micelle fractions. As a consequence, there was significant interaction between casein micelle and milk serum, causing cloudiness of SerP in the liquid-state 31P-NMR spectrum of milk. Casein composition influenced the SerP-MCP interaction in micellar structure. Shape changing of the SerP peak was discussed in connection with beta-casein-release phenomenon.     

Evaporative concentration of skimmed milk: Effect on casein micelle hydration,composition, and size

Understanding the effect of evaporative concentration on casein micelle composition is of high importance for milk processing. Alterations to the hydration, composition and size of casein micelles were investigated in skimmed milk evaporated to concentrations of 12–45% total solids content. The size of casein micelles was determined by dynamic light scattering, and the water content and composition determined by analysis of supernatants and pellets obtained by ultracentrifugation. The mass balance and hydration results showed that during the evaporation process, while micelles were dehydrated, water was removed preferentially from the serum. The amount of soluble casein and calcium in the serum decreased as a function of increasing solids content, indicating a shift of these components to the micelles. The formation of a small proportion of micelle aggregates at high concentrations appeared dependent on the time kept at these concentrations. Upon redilution with water, casein micelles were immediately rehydrated and aggregates were broken up in a matter of minutes. Soluble calcium and pH returned to their original state over a number of hours; however, only a small percentage of original soluble casein returned to the serum over the 5 h period investigated. These results showed that casein micelles are significantly affected by evaporative concentration and that the alterations are not completely and rapidly reversible.     

The dominating effect of ionic strength on the heat-induced denaturation and aggregation of β-lactoglobulin in simulated milk ultrafiltrate

The denaturation/aggregation behaviour of heated (78 °C, 10 min) β-lactoglobulin (1%, w/w) was examined as a function of heating pH (5.0–7.0), in the presence of different salts. Heating β-lactoglobulin in the presence of calcium (5 mm) significantly increased the level of aggregated protein at most heating pH values, compared to heating in water or sodium chloride (100 mm). Heating β-lactoglobulin in the presence of calcium (5 mm) and phosphate (5 mm), resulted in similar denaturation levels in the pH range 5.0–5.8 as in the presence of calcium (5 mm) alone but reduced denaturation in the pH range 6.0–7.0, probably due to the formation of insoluble calcium phosphate. The addition of NaCl (100 mm) counteracted the aggregation promoting properties of the calcium and calcium/phosphate systems. Heating β-lg in a simulated milk ultrafiltrate solution was similar to heating in NaCl alone. This suggested that Ca²⁺ effects alone may not explain the heat-induced denaturation/aggregation behaviour of β-lactoglobulin in milk whey systems.     

Kinetics of Protein Extraction in Reverse Micelle

Sodium bis (2-ethylhexyl) sulfosuccinate (AOT)-sodium dodecyl sulfate (SDS)/isooctane-octanol reverse micelle extraction was tested an efficient and effective approach to separate peanut protein from full-fat peanut powder. Here, important kinetic factors including pH, ion strength, and temperature were studied during reverse micelle backward extraction. The extraction conditions were obtained by response surface experiments as follows: pH 7.5, ion concentration 1.1 mol/L at temperature 35°C. Under these optimum extraction conditions, the extraction rate of protein reached 79.03%. A model on the kinetic partitioning of peanut protein was also developed. The backward extraction in this reverse micelle system was controlled by interfacial resistance instead of diffusion resistance in reverse micelle and aqueous phase with the total mass transfer rate of 0.8×10^?5 m³·s^?1. A two-film theory may be the mechanism for flat interface. Results of mass transfer process are helpful for creating an reverse micelle extraction process, and used for purification of peanut proteins, promoting the development of food industry.     

Heat-induced changes in the calcium sensitivity of caseins

The calcium sensitivity of Na caseinate prepared from a serum protein-free casein micelle dispersion in synthetic milk ultrafiltrate, containing 4.8%, w/v, lactose and 5 mmol L⁻¹ urea and heated at 130°C for 0–25 min decreased with heating. It is proposed that an increase in the net negative charge of the caseinates, due to heat-induced degradation of lysine and arginine, is responsible for the enhanced calcium stability of heated caseins. The Maillard reaction and urea–protein interactions appear to play an important role in the increased stability of heated caseinate towards calcium. The effect of protein charge on the heat stability of milk protein systems (Na caseinate 2.5%, w/v, protein in milk ultrafiltrate) at 140°C was investigated by chemical modification of the caseinate prior to assessment of heat stability. Heat stability increased with the modification of lysine, arginine and carboxyl residues. The increased heat stability of Na caseinate with modified lysine and arginine residues may be due to an increase in the net negative charge on the caseinate, while the increased stability of caseinate with modified carboxyl residues may be related to a reduction in heat-induced crosslinking of protein.     

In-vitro and in-vivo dephosphorylation of rapeseed meal by means of phytate-degrading enzymes derived from Aspergillus Niger

A meal of ‘double low’ rapeseed (var ‘Jantar’) was subjected to phytate hydrolysis using enzyme preparations derived from a mycelium of Aspergillus niger which contained phytase (EC 3.1.3.8) and acid phosphatase (EC 3.1.3.2) activities. The complete conversion of myo-inositol hexa- and pentaphosphates present in rapeseed meal to lower phosphate esters of myo-inositol was accomplished at 40°C, a pH value of 4.5, phytase dosage (in phytase units (PhytU)) 0.1 PhytU g^?1 accompanied by acid phosphatase activity 37.1 units g^?1, in 1 h. Under these conditions, complete dephosphorylation was observed in 4 h. Decreasing the pH value to 3.0 caused a rise in the amount of inorganic phosphorus released, while increasing to 5.5 resulted in substantial reduction in the reaction rate. Purification of phytase to a specific activity 0.375 PhytU mg^?1 of protein exhibited a negative influence upon the yield of rapeseed dephosphorylation. The substitution of calcium phosphate for a preparation of phytase in feed containing rapeseed meal did not cause significant differences in the body weight gain or in tibia mineralisation of broilers (Gains galus, ‘Astra B’).