Rennet-induced gelation of calcium and phosphate supplemented skim milk subjected to CO2 treatment

A Doehlert design was performed to study the effect of calcium and phosphate supplementation at 0 to 25 mmol/kg and 0 to 16 mmol/kg, respectively, on the rennet gelation of reconstituted skim milk subjected to pH-reversible CO(2) acidification. Supplemented reconstituted skim milk samples were acidified to pH 5.80 by the addition of CO(2) under pressure and depressurized under vacuum to restore the initial pH value. The second-order polynomial models satisfactorily predicted the effect of salt addition on the micellar molar Ca:P ratio and the average diameter of the casein micelles, whereas only trends were used in the analysis of the rennet-clotting behavior of salt-supplemented, CO(2)-treated milk. Whether added Ca was the most determinant factor on the micellar molar Ca:P ratio, added Pi (a mixture of Na(2)HPO(4) and NaH(2)PO(4)) was the most determinant factor on the other responses studied, and its effect was most pronounced when Ca was simultaneously added. By comparison with control samples, changes observed in this study were essentially due to salt supplementation and not to the CO(2) treatment. Therefore, this CO(2) treatment could be considered as an entirely reversible treatment rather than only pH-reversible, and predictions might be applied to untreated milk. In the case of Ca-supplemented milk, the micellar molar Ca:P ratio increased, the average micellar diameter decreased, and the rennet-clotting properties were improved, whereas opposite effects were observed upon Pi supplementation. Since modification of the micellar molar ratio is the result of change in the chemical composition of micellar calcium phosphate, the effect of calcium and phosphate supplementation on the rennet clotting of milk was found to be also dependent on the nature of the interaction between caseins and colloidal calcium phosphate.     

Mineral and casein equilibria in milk: effects of added salts and calcium-chelating agents

We have investigated the effects of adding a range of mineral salts and calcium-chelating agents on the distribution of casein and minerals between the non-pelleted and pelleted phases of milk obtained upon centrifugation at 78000 g for 90 min. Adding CaCl2 or mixtures of NaH2PO4 and Na2HPO4 to reconstituted skim milk (90 g milk solids/kg) at pH 6.65 increased both pelleted casein and pelleted calcium phosphate. Opposite effects were obtained by adding citrate or EDTA. The change in pelleted calcium phosphate was not simply related to casein release from the micelle. Upon adding 5 mmol EDTA/kg milk, 20% of the pelleted Ca, 22% of the pelleted phosphate and 5% of the micellar casein were removed. Increasing the concentration of EDTA to 10 mmol/kg milk decreased the pelleted Ca by 44% and the pelleted phosphate by 46%, and caused 30% of the micellar casein to be released. The effects of adding phosphate, citrate or EDTA at pH 6.65, followed by the addition of CaCl2, demonstrated the reversibility of the dissolution and formation of the micellar calcium phosphate. There were limits to this reversibility that were related to the amount of colloidal calcium phosphate removed from the casein micelles. Adding CaCl2 to milk containing > or = 20 mmol EDTA or > or = 30 mmol citrate/kg milk did not result in complete reformation of casein micelles. Light-scattering experiments confirmed that the dissolution of moderate amounts of colloidal calcium phosphate had little effect on micellar size and were reversible, while the dissolution of larger amounts of colloidal calcium phosphate resulted in large reductions in micellar size and was irreversible.     

Influence of calcium salt supplementation on calcium equilibrium in skim milk during pH cycle

Calcium is a mineral essential for humans, especially for bone constitution. Yet most of the worldwide population does not satisfy their Ca needs. Hence, Ca supplementation is of major importance, even in western countries where some specific populations at risk do not satisfy the recommended daily intake of Ca. More than 70% of dietary Ca comes from dairy products. Calcium supplementation of naturally Ca-rich sources such as skim milk is then of special interest. To our knowledge, few data are available concerning milk Ca (MC) supplementation of milk, particularly when followed by pH cycle. In this paper, MC supplementation is studied and compared with Ca chloride (CC) supplementation as a well-known source of Ca. The effect of Ca salt supplementation followed by pH cycle was studied in reconstituted skim milk. Calcium supplementation was carried out with CC and MC at 25 mmol of Ca/kg of skim milk. Ionized Ca concentration and turbidity variations were followed in situ by Ca ion selective electrode and turbidimetry using light reflection. From normalized data on ionized Ca concentration and turbidity vs. pH, it appeared that hysteresis areas were smaller for CC-supplemented milk, whereas unsupplemented milk and MC-supplemented milk behaved similarly. For these 3 dairy systems, pH cycles to pH 5.0 led to a larger hysteresis area than pH cycles to pH 5.5. The shrinkage of the hysteresis area could be interpreted as a reinforcement of casein micelles with Ca ions over the pH cycle.     

The use of sedimentation field flow fractionation and photon correlation spectroscopy in the characterization of casein micelles

Sedimentation Field Flow Fractionation (SdFFF) was combined with Photon Correlation Spectroscopy (PCS), to characterize changes in the structure of the colloidal particles of reconstituted skim milk of diameter >50 nm (aggregates of casein and calcium phosphate known as casein micelles) with the changes in partitioning (with the addition of salt) of calcium (Ca), inorganic phosphate (Pi) and casein between the serum and colloidal phases of the milk. The number weighted particle size distributions are determined. These are well represented by a log-normal distribution. Methods are presented for estimating the relative contributions of scattering and absorbance to the SdFFF detector signal and for taking both into account when analysing SdFFF data. The values found for the effective density of the casein micelles were in good agreement with the literature and ranged from (1.06-1.08 g cm(-3)) according to the composition of micelles. The changes in the scattering intensity as determined by PCS correlated with the changes in the particle composition. Although the concentrations of colloidal calcium phosphate (CCP) (1.1-3.5 g/kg milk) and micellar casein (18.1-27.2 g/kg milk) varied considerably only small changes in the size distribution of particles >50 nm diameter were observed except for milk to which 30 mmol Pi+10 mmol Ca/kg milk had been added where the particle size distribution shows a swelling of the particles consistent with a lower than expected value for the particle density. These observations suggest that the micelles have the ability to both lose (depleted micelles) and accommodate (enriched micelles) more casein, calcium and inorganic phosphate in their interior, thus confirming the model of the micelles which postulates an open structure allowing freedom of movement of casein and small ions.     

Zinc binding in bovine milk

About 90% of the Zn in bovine skim milk was sedimented by ultracentrifugation at 100,000 g for 1 h. About half of the non-sedimentable Zn was non-dialysable, indicating that it was associated with protein, probably non-sedimented casein micelles. Casein micelles incorporated considerable amounts of Zn added to skim milk as ZnCl2, and at Zn concentrations greater than or equal to 16 mM coagulation of casein micelles occurred. Ca was displaced from casein micelles by increasing ZnCl2 concentration and approximately 40% of micellar Ca was displaced by 16 mM-ZnCl2. Micellar Zn, Ca and Pi were gradually rendered soluble as the pH of milk was lowered and at pH 4.6 greater than 95% of the Zn, Ca and Pi were non-sedimentable. These changes were largely reversible by readjustment of the pH to 6.7. About 40% of the total Zn in skim milk was non-sedimentable at 0.2 mM-EDTA and most of the remainder was gradually rendered soluble by EDTA over the concentration range 1-50 mM. This indicates that there are two distinct micellar Zn fractions. No micellar Ca or Pi was solubilized at EDTA concentrations up to 1.0 mM, indicating that both colloidal calcium phosphate (CCP) and casein micelles remained intact under conditions where the more loosely bound micellar Zn fraction dissolved. Depletion of casein micelles of colloidal Ca and Pi by acidification and equilibrium dialysis resulted in removal of Zn, and in colloidal Pi-free milk non-dialysable Zn was reduced to 1.2 mg/l (approximately 32% of the original Zn). Thus, approximately 32% of the Zn in skim milk is directly bound to caseins, while approximately 63% is associated with CCP. Over 80% of the Zn in colloidal Pi-free milk was rendered soluble by 0.2 mM-EDTA, indicating that the casein-bound Zn is the loosely bound Zn fraction in casein micelles. A considerable fraction of the Zn in acid whey (pH 4.6) co-precipitated with Ca and Pi on raising the pH to 6.7 and heating for 2 h at 40 degrees C, indicating that insoluble Zn phosphate complexes form readily under these conditions. Studies on dialysis of milk against water, or dilution of milk or casein micelles with water, showed that CCP and its associated Zn is very stable and dissolves only very slowly at pH 6.6. The nature of Zn binding in casein micelles may help to explain the lower nutritional bioavailability of Zn in bovine milk and infant formulae compared with human milk.     

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.     

Effects of reverse CO2 acidification cycles,calcium supplementation,pH adjustment and chilled storage on physico-chemical and rennet coagulation properties of reconstituted low- and medium-heat skim milk powders

The reversibility extent of one and two reverse CO₂ acidification cycles on the physico-chemical and rennet coagulation properties of milks reconstituted from low- (LH) or medium- (MH) heat skim powder, enriched or not with calcium and pH adjusted or not was investigated. The ionized calcium concentration, buffering properties and average casein micelle size of untreated and CO₂-treated milks were evaluated before and after a chilled storage for 2 days. The ionized calcium concentration and buffering properties have been modified by the CO₂-treatment, particularly after a second CO₂-cycle. These modifications were highly dependent on the initial milk properties and chilled storage. Inversely, the average casein micelle size was not significantly changed. In addition, the rennet-clotting behaviour checked by near infrared spectroscopy (NIR-S) and rheology (SAOR) indicated the main factors responsible for changes in the casein micelles environment and dynamic casein micellar calcium phosphate reorganization, especially after two CO₂-cycles. A single CO₂-cycle induced a better rennetability for non Ca-enriched milk reconstituted from MH-powder. A second CO₂-cycle was particularly efficient to improve Ca-enriched pH-adjusted milks.     

Acid Milk Gel Formation as Affected by Total Solids Content

Reconstituted skim milk with varying concentrations of total solids was coagulated using glucono-δ-lactone (GDL). Microscopic, turbidimetric and rheological procedures were used to examine mineral solubilization, buffering capacity, casein dissociation and micellar solvation during gelation. Total solids of the milk affected pH of the onset of gelation attributable to differences in colloïdal calcium phosphate in the casein particles during acidification. Firmness and elasticity of the resulting gel increased with total solids from a more direct contribution of dry matter during the last stage of acid milk gel formation.     

High pressure effects on the colloidal calcium phosphate and the structural integrity of micellar casein in milk. Part 1. High pressure dissolution of colloidal calcium phosphate in heated milk systems

Results of this study confirm that high temperature (118°C/15 min) and high pressure (400 MPa/5 min) processing of skim milk, skim milk ultrafiltration and ultracentrifugation fractions, and model milk salt solutions cause dramatic shifts in their colloidal and soluble Ca phospate equilibrium that affect their pH, dissolved Ca content, turbidity, and casein micelle microstructure. The relations between high temperature and high pressure processing-induced changes in the colloidal and soluble Ca phosphate equilibrium were evaluated in raw, pasteurized, and high temperature treated skim milk, ultrafiltration retentate and permeate of pasteurized skim milk, clear ultracentrifugation infranatant of pasteurized skim milk, and synthetic milk ultrafiltrates with and without lactose or Ca. The magnitude of the pH and dissolved Ca shifts caused by high temperature and high pressure processing was a function of casein micelle concentration. Ultrafiltration permeate exhibited the most drastic shifts in pH and dissolved Ca contents due to high temperature and high pressure processing. Although high temperature processing reduced the pH of ultrafiltration permeate from 6.59 to 6.03 and the dissolved Ca from 100% to 58%, high pressure processing reversed both of these changes. These changes in high temperature and high pressure processed milk, milk fractions, and model milk salt solutions were related to microstructural changes in the casein micelles as revealed by electron microscopy.     

Influence of stabilizers on the salt balance and pH of buffalo milk and its concentrate

The influence of three different concentrations (0.05%, 0.10% and 0.15%) of two stabilizers, disodium phosphate and trisodium citrate on the salt balance (concentration and molar ratios of salt constituents in the dissolved phase) and pH of buffalo milk and its 2:1 concentrate was determined. The disodium phosphate caused a significant shift in all the salt constituents (calcium, magnesium, phosphate and citrate) from the dissolved to the colloidal phase while the trisodium citrate produced a significant shift from the colloidal to the dissolved phase. Further, the phosphate caused a uniform decrease in the molar ratios of Ca/P and (Ca+Mg)/(P+Cit.) in the dessolved phase, while the citrate produced only a small and non-significant effect. Both salts caused a significant increase in pH which was progressive with increase in the concentration of added salts. Therefore, the primary effect of stabilizers in stabilizing or destabilizing the milk is a consequence of their influence on the pH and not on the mineral equilibrium of milk.     

Serum protein and casein concentration: effect on pH and freezing point of milk with added CO2

The objective of this study was to determine the effect of protein concentration and protein type [i.e., casein (CN) and serum protein (SP)] on pH (0 degree C) and freezing point (FP) of skim milk upon CO2 injection at 0 degree C. CN-free skim milks with increasing SP content (0, 3, and 6%) and skim milks with the same SP content (0.6%) but increasing CN content (2.4, 4.8, and 7.2%) were prepared using a combination of microfiltration and ultrafiltration processes. CO2 was injected into milks at 0 degree C using a continuous flow carbonation unit (230 ml/min). Increasing SP or CN increased milk buffering capacity and protein-bound mineral content. At the same CO2 concentration at 0 degree C, a milk with a higher SP or a higher CN concentration had more resistance to pH change and a greater extent of FP decrease. The buffering capacity provided by an increase of CN was contributed by both the CN itself and the colloidal salts solublized into the serum phase from CN upon carbonation. Skim milks with the same true protein content (3%), one with 2.4% CN plus 0.6% SP and one with 3% SP, were compared. At the same true protein content (3%), increasing the proportion of CN increased milk buffering capacity and protein-bound mineral content. Milk with a higher proportion of CN had more resistance to pH change and a greater extent of FP decrease at the same carbonation level at 0 degree C. Once CO2 was dissolved in the skim portion of a milk, the extent of pH reduction and FP depression depended on protein concentration and protein type (i.e., CN and SP).     

Effect of increasing the colloidal calcium phosphate of milk on the texture and microstructure of yogurt

The effect of increasing the colloidal calcium phosphate (CCP) content on the physical, rheological, and microstructural properties of yogurt was investigated. The CCP content of heated (85°C for 30 min) milk was increased by increasing the pH by the addition of alkali (NaOH). Alkalized milk was dialyzed against pasteurized skim milk at approximately 4°C for 72 h to attempt to restore the original pH and soluble Ca content. By adjustment of the milk to pH values 7.45, 8.84, 10.06, and 10.73, the CCP content was increased to approximately 107, 116, 123, and 128%, respectively, relative to the concentration in heated milk. During fermentation of milk, the storage modulus (G′) and loss tangent values of yogurts were measured using dynamic oscillatory rheology. Large deformation rheological properties were also measured. The microstructure of yogurt was observed using fluorescence microscopy, and whey separation was determined. Acid-base titration was used to evaluate changes in the CCP content in milk. Total Ca and casein-bound Ca increased with an increase in the pH value of alkalization. During acidification, elevated buffering occurred in milk between pH values 6.7 to 5.2 with an increase in the pH of alkalization. When acidified milk was titrated with alkali, elevated buffering occurred in milk between pH values 5.6 to 6.4 with an increase in the pH of alkalization. The high residual pH of milk after dialysis could be responsible for the decreased contents of soluble Ca in these milks. The pH of gelation was higher in all dialyzed samples compared with the heated control milk, and the gelation pH was higher with an increase in CCP content. The sample with highest CCP content (128%) exhibited gelation at very high pH (6.3), which could be due to alkali-induced CN micellar disruption. The G′ values at pH 4.6 were similar in gels with CCP levels up to 116%; at higher CCP levels, the G′ values at pH 4.6 greatly decreased. Loss tangent values at pH 5.1 were similar in all samples except in gels with a CCP level of 128%. For dialyzed milk, the whey separation levels were similar in gels made from milk with up to 107% CCP but increased at higher CCP levels. Microstructure of yogurt gels made from milk with 100 to 107% CCP was similar but very large clusters were observed in gels made from milk with higher CCP levels. By dialyzing heated milk against pasteurized milk, we may have retained some heat-induced Ca phosphate on micelles that normally dissolves on cooling because, during dialysis, pasteurized milk provided soluble Ca ions to the heated milk system. Yogurt texture was significantly affected by increasing the casein-bound Ca (and total Ca) content of milk as well as by the alkalization procedure involved in that approach.     

Effects of high pressure and heat treatment on the mineral balance of goats' milk

Changes in the distribution of minerals in skim goats' milk by high pressure (400 MPa) and/or heat (85 degrees C for 30 min) treatment have been studied. Heat treatment caused reduced solubility of the calcium, magnesium and phosphorus, and this increased with the severity of heating. In contrast, high pressure released different levels of micellar elements into the soluble phase without causing appreciable changes in pH or ionic calcium concentration. The levels of soluble salts returned to their original values when the heated samples were subjected to high pressure. However, heating pressurized milk resulted in concentrations of soluble minerals that were lower than in control milks, and close to values found in heated milks. The salt balance in goats' milk was less affected by high pressure treatment at 75 degrees C than was that of cows' milk. These results are discussed in relation to the effects of high pressure and heat treatment on mineral equilibrium and micellar structure.     

Changes in the proportions of soluble and insoluble calcium during the ripening of cheddar cheese

In cheese, the concentration and form of residual Ca greatly influences texture. Two methods were used to determine the proportions of soluble (SOL) and insoluble (INSOL) Ca in Cheddar cheese during 4 mo of ripening. The first method was based on the acid-base buffering curves of cheese and the second was based on the extraction of the aqueous phase ("juice") of cheese under high pressure and determining the concentration of SOL Ca in the juice using atomic absorption spectroscopy. When cheese was acidified there was a strong buffering peak at pH approximately 4.8, which was due to the solubilization of residual colloidal calcium phosphate (CCP) of milk that remained in cheese as INSOL Ca phosphate. The area of this buffering peak in cheese was expressed as a percentage of the original area of this peak in milk and was used to estimate the concentration of residual INSOL Ca phosphate in cheese. There were no significant differences between the 2 methods. The proportions of INSOL Ca in cheese decreased from approximately 73 to approximately 58% between d 1 and 4 mo. These methods will be useful techniques to study the role of Ca in cheese texture and functionality.     

INFLUENCE OF HOMOGENIZATION ON THE HEAT STABILITY AND SALT BALANCE OF BUFFALO MILK AND ITS 1:2 CONCENTRATE

The influence of homogenization (two stage) on heat stability (determined as heat coagulation time at 130°) as well as the salt balance of buffalo milk and its 1:2 concentrate was determined. Homogenization had no significant influence on the heat stability and salt balance (molar ratio of Ca + Mg to P + Cit. in the dissolved phase) of fluid milk. However, during concentration both the heat coagulation time and salt balance of homogenized milk were affected significantly. Homogenization caused a significant decrease in the dissolved phosphate but had no significant effect on calcium and magnesium and this resulted in a significant increase in the molar ratio of Ca + Mg/P + Cit. This disruption in the salt equilibrium caused the destabilization of homogenized concentrated milk.     

浓缩酪蛋白胶束成分的影响因素及其在奶酪生产中的应用

目前,采用膜过滤技术可从脱脂奶中分离酪蛋白,随后通过浓缩、杀菌、干燥等工艺获得浓缩酪蛋白胶束。对浓缩酪蛋白胶束成分的影响因素及其在奶酪生产中的应用进行综述,发现膜过滤期间的pH值、温度和洗滤条件均会影响浓缩酪蛋白胶束的成分,使其具有不同浓度的酪蛋白、乳清蛋白、乳糖以及钙。而且可以利用浓缩酪蛋白胶束标准化原奶,从而制备成分和品质一致的奶酪;也可以利用不同成分的浓缩酪蛋白胶束获得不同的原奶组合物,从而制备所需品质和功能的奶酪。总之,在奶酪生产过程中添加浓缩酪蛋白胶束能够影响奶酪的成分、质地以及风味等,但通过调整膜过滤和奶酪生产的工艺参数可以解决这些问题。未来还需获得一种经济有效的方式来保存浓缩酪蛋白胶束,赋予其更长的保质期,良好的凝乳酶凝乳特性,从而保证奶酪的品质和产量。     

Impact of source and level of calcium fortification on the heat stability of reconstituted skim milk powder

Calcium enrichment of food and dairy products has gained interest with the increased awareness about the importance of higher calcium intake. Calcium plays many important roles in the human body. Dairy products are an excellent source of dietary calcium, which can be further fortified with calcium salts to achieve higher calcium intake per serving. However, the addition of calcium salts can destabilize food systems unless conditions are carefully controlled. The effect of calcium fortification on the heat stability of reconstituted skim milk was evaluated, using reconstituted skim milks with 2 protein levels: 1.75 and 3.5% (wt/wt) prepared using low and high heat powders. Calcium carbonate, phosphate, lactate, and citrate were used for fortification at 0.15, 0.18, and 0.24% (wt/wt). Each sample was analyzed for solubility, heat stability, and pH. The addition of phosphate and lactate salts lowered the pH of milk, citrate did not have any major effect, and carbonate for the 1.75% protein samples increased the pH. In general, changes in solubility and heat stability were associated with changes in pH. Calcium addition decreased the solubility and heat stability. However, interestingly, the presence of carbonate salt greatly increased the heat stability for 1.75% protein samples. This is due to the neutralizing effect of calcium carbonate when it goes into solution. The results suggested that the heat stability of milk can be affected by the type of calcium salt used. This may be applied to the development of milk-based calcium enriched beverages.     

Effects of pH,calcium-complexing agents and milk solids concentration on formation of soluble protein aggregates in heated reconstituted skim milk

The proportion of protein present in the supernatant after centrifugation and the formation of soluble protein aggregates in heated (90 °C for 10 min) reconstituted skim milk were investigated as a function of (a) pH, (b) milk concentration 9–21%, w/w (milk solids non-fat, MSNF) and (c) addition of calcium-complexing agents (orthophosphate or EDTA). Compositional changes in milk, resulting from pH adjustment or salt addition, altered the distribution of caseins between the serum and micellar phases of milk prior to heating, and influenced the amount and composition of soluble aggregates formed during heat treatment. Although milk pH was a good predictor of the aggregation behaviour of the proteins during heating of milk of different solids concentrations, neither the proportion of protein in the supernatant prior to heating nor the pH alone could predict the aggregation behaviour when the composition was adjusted with mineral salts.     

Properties of casein micelles cross-linked by transglutaminase

Factors affecting the cross-linking of milk proteins by transglutaminase (TGase) were studied. Cross-linking of caseins in bovine skim milk was optimal over a very wide pH range. The role of micellar calcium phosphate (MCP) in maintaining the integrity of TGase-treated casein micelles was studied by incubating skim milk with 0.01% (w/v) TGase at 30°C for 1–24 h, followed by removal of MCP from untreated or TGase-treated milk by acidification and dialysis. The protein content and profile of the samples were determined by Kjeldahl and SDS-PAGE, respectively. Whey proteins in unheated milk were not susceptible to TGase-induced cross-linking. The higher level of sedimentable protein in MCP-free TGase-treated milk than in MCP-free control milk indicated that TGase treatment partially prevented disintegration of the micelle on removal of MCP, probably due to extensive intramicellar TGase-induced cross-linking of casein molecules which led to the formation of sedimentable covalently bonded casein aggregates.     

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.