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.     

Effect of partial whey protein depletion during membrane filtration on thermal stability of milk concentrates

Membrane filtration technologies are widespread unit operations in the dairy industry, often employed to obtain ingredients with tailored processing functionalities. The objective of this work was to better understand the effect of partial removal of whey proteins by microfiltration (MF) on the heat stability of the fresh concentrates. The micellar casein concentrates were compared with control concentrates obtained using ultrafiltration (UF). Pasteurized milk was microfiltered (80 kDa polysulfone membrane) or ultrafiltered (30 kDa cellulose membrane) without diafiltration (i.e., no addition of water) to 2× and 4× concentration, based on volume reduction. The final concentrates showed no differences in pH, casein micelle size, or mineral concentration in the serum phase. The micellar casein retentates (obtained by MF) showed a 20 and 40% decrease in whey protein concentration compared with the corresponding UF milk protein concentrates for 2× and 4× concentration, respectively. The heat coagulation time decreased with increasing protein concentration, regardless of the treatment; however, MF retentates showed a higher thermal stability than the corresponding UF controls. The average diameter for casein micelles increased after heating in UF but not MF concentrates. The turbidity (measured by light scattering) increased after heating, but to a higher extent for UF retentates than for MF retentates at the same protein concentration. It was concluded that the reduced amount of whey protein in the MF retentates caused a significant increase in the heat stability compared with the corresponding UF retentates. This difference was not due to ionic composition differences or pH, but to the type and amount of complexes formed in the serum phase.     

Impact of modifications in acid development on the insoluble calcium content and rheological properties of Cheddar cheese

Cheddar cheese was made from milk concentrated by reverse osmosis (RO) to increase the lactose content or from whole milk. Manufacturing parameters (pH at coagulant addition, whey drainage, and milling) were altered to produce cheeses with different total Ca contents and low pH values (i.e., <5.0) during ripening. The concentration of insoluble (INSOL) Ca in cheese was measured by cheese juice method, buffering by acid-base titration, rheological properties by small amplitude oscillatory rheometry, and melting properties by UW-Melt Profiler. The INSOL Ca content as a percentage of total Ca in all cheeses rapidly decreased during the first week of aging but surprisingly did not decrease below approximately 41% even in cheeses with a very low pH (e.g., ∼4.7). Insoluble Ca content in cheese was positively correlated (r = 0.79) with cheese pH in both RO and nonRO treatments, reflecting the key role of pH and acid development in altering the extent of solubilization of INSOL Ca. The INSOL Ca content in cheese was positively correlated with the maximum loss tangent value from the rheology test and the degree of flow from the UW-Melt Profiler. When cheeses with pH <5.0 where heated in the rheometer the loss tangent values remained low (<0.5), which coincided with limited meltability of Cheddar cheeses. We believe that this lack of meltability was due to the dominant effects of reduced electrostatic repulsion between casein particles at low pH values (<5.0).     

Buffering curves of ideal whey fractions obtained from a cascade membrane separation process

Skimmed milk was fractionated via a cascade system: Graphic plotting of microfiltration (MF); ultrafiltration (UF); nanofiltration (NF); and reverse osmosis (RO). The buffering curves of each fraction were studied over the pH range 4–7. Depending on their composition, the individual permeate streams showed different buffering capacity values and pH ranges where the buffering occurred. The concentration of active buffering substances in the permeates decreased (in mmol/L) from ~26.6 (MF) to ~17.4 (UF) to 1.39 (NF) to 0.07 (RO). Contributions to the total buffering capacity for MF permeate, which represents the serum phase of milk, were ~37% from whey proteins and ~63% from milk salts (especially citrates, phosphates and carbonates) including lactose and water.     

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.     

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.     

Cold ultrafiltered or microfiltered milk retentates: A systematic comparison of the effects of compositional differences on their gelation functionality

The colloidal stability of casein micelles suspensions prepared using ultrafiltration (UF) and microfiltration (MF) was studied by testing acid- and rennet-induced destabilization. Skim milk and 4× (based on volume reduction) concentrates were obtained by processing under similar conditions, at temperatures below 10°C. Concentrates were subjected to different levels of diafiltration (DF), resulting in samples with comparable casein volume fractions but different amounts of proteins and ions in the serum phase. The novelty of the work is the systematic comparison of MF and UF concentrates of similar history. More specifically, concentrates similar in ionic composition but with or without serum proteins were compared, to evaluate whether whey proteins and β-casein depletion from the micelles will play a role in the processing properties, or whether these are affected solely by the ionic balance. Microfiltered micelles' apparent diameter decreased by about 50 nm during the specific hydrolysis of κ-casein by chymosin, whereas those in skim milk control showed a decrease of about half that size. All concentrates subjected to extensive DF showed smaller hydrodynamic diameters, with reductions of ∼18 and 13 nm for MF and UF, respectively. Highly diafiltered UF retentates showed a delayed onset of rennet-induced gelation, due to low colloidal calcium, compared with other samples. Low-diafiltered samples showed weak storage modulus (∼1 Pa) after 60 min of onset of gelation. In addition, onset pH increased with diafiltration to ∼5.8 for UF and ∼6 for MF in high-diafiltered samples. These results clearly demonstrated that the functional properties of casein micelles change during membrane concentration, and this cannot be solely attributed to changes in ionic equilibrium.     

Effect of Tetrasodium Pyrophosphate on the Physicochemical Properties of Yogurt Gels

The effect of tetrasodium pyrophosphate (TSPP) on the properties of yogurt gels was investigated. Various concentrations (0.05 to 0.2%) of TSPP were added to preheated (85°C for 30 min) reconstituted skim milk, which was readjusted to pH 6.50. Milk was inoculated with 2% starter culture and incubated at 42°C until the pH reached 4.6. Acid-base buffering profiles of milk and total and soluble calcium levels were measured. Turbidity measurements were used to indicate changes in casein dispersion. Storage modulus (G′) and loss tangent (LT) values of yogurts were monitored during fermentation using dynamic oscillatory rheology. Large deformation properties of gels were also measured. Microstructural properties of yogurt were observed using fluorescence microscopy. The addition of TSPP resulted in the disappearance of the buffering peak during acid titration at pH ∼5.1 that is due to the solubilization of colloidal calcium phosphate (CCP), and a new peak was observed at lower pH values (pH 4.0-4.5). The buffering peak at pH 6.0 during base titration virtually disappeared with addition of TSPP and a new peak appeared at pH ∼4.8. The addition of TSPP reduced the soluble Ca content of milk and increased casein-bound Ca values. The addition of up to 0.125% TSPP resulted in a reduction in turbidity because of micelle dispersion but at 0.15%, turbidity increased and these samples exhibited a time-dependent increase in turbidity because of aggregation of casein particles. Gels made with 0.20% TSPP were very weak and had a very high gelation pH (6.35), probably due to complete dispersion of the micelle structure in this sample. The LT value of gels at pH 5.1 decreased with an increase in TSPP concentration, probably due to the loss of CCP with the addition of TSPP. The G′ values at pH 4.6 of gels made with ≤0.10% TSPP were not significantly different but the addition of ≥0.125% TSPP significantly decreased G′ values. The addition of 0.05 to 0.125% TSPP to milk resulted in a reduction in the yield stress values of yogurt compared with yogurt made without TSPP. Greater TSPP levels (>0.125%) markedly reduced the yield stress values of yogurt. Lowest whey separation levels were observed in yogurts made with 0.10% TSPP. High TSPP levels (>0.10%) greatly increased the apparent pore size of gels. Addition of very low levels of TSPP to milk for yogurt manufacture may be useful in reducing the whey separation defect, but at TSPP concentrations ≥0.125% very weak gels were formed.     

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 acidification on physico-chemical characteristics of buffalo milk: A comparison with cow’s milk

Composition and physico-chemical properties of buffalo and cow milks were compared at their initial pH and during acidification. As compared to cow milk, buffalo milk was richer in fat, lactose, protein (especially caseins) and minerals such as calcium, magnesium and inorganic phosphate. Along with these differences of major components, the capacity of milk to be acidified (named buffering capacity) was higher for buffalo milk than for cow milk. The precipitation/aggregation of caseins at their isoelectric pH, solubilization of calcium and inorganic phosphate and decrease in hydration of casein as a function of decrease in pH were significant for both milks. For both species, these molecular changes were qualitatively similar but quantitatively different. These quantitative differences during acidification were related to the differences of composition between the milks.     

Observations of casein micelles in skim milk concentrate by transmission electron microscopy

The microstructure of casein micelles in ultrafiltrated (UF) skim milk concentrate at pH 6.5 and 5.8 was investigated by transmission electron microscopy using three different preparation methods. The volume fraction of the casein micelles in the UF concentrate was 62.8% (v/v) at pH 6.5 and fixation by glutaraldehyde revealed the close packing of micelles in the UF concentrate as well as a higher degree of micelle aggregation at pH 5.8. No details of the microstructure of the micellar surface or core could, however, be observed. Freeze-fracture of cryoprotected, i.e. glycerol, UF concentrate on the other hand, exposed the finer structures of the micellar core but no pH dependent differences were observed. As cryoprotection includes a dilution of the sample with glycerol, the packing of the micelles in the UF concentrate could not be observed. Undiluted UF concentrate exposed to rapid freezing using a propane jet followed by freeze-fracture exhibited development of ice crystals but rough areas on the micrographs were identified as fractured casein micelles. The micellar core appeared rougher and differences in the micellar core microstructure due to changing pH could be observed when this preparation method was used.     

Effect of insoluble calcium concentration on rennet coagulation properties of milk

Rennet-induced gels were made from milk acidified to various pH values or milk at pH 6.0 that had added EDTA. The objective was to examine the effect of removing insoluble Ca (INS Ca) from casein micelles (CM) on rennet gelation properties. For the pH trial, diluted lactic acid was added to reconstituted skim milk to decrease the pH to 6.4, 6.0, 5.8, 5.6, and 5.4. For the EDTA trial, EDTA was slowly added (0, 2, 4, and 6 mM) to reconstituted skim milk, and the final pH values were subsequently adjusted to pH 6.0. Dynamic low amplitude oscillatory rheology was used to monitor gel development. The Ca content of CM and rennet wheys made from these milks was measured using inductively coupled plasma spectroscopy. The INS Ca content of milk was altered by the acidification pH values or level of EDTA added. In all samples, the storage modulus (G′) exhibited a maximum (GM), with a decrease in G′ during longer aging times. Gels made at pH 6.4 had higher GM compared with gels made at pH 6.7 probably due to the reduction in electrostatic repulsion, whereas the INS Ca content only slightly decreased. The highest GM value of gels was observed at pH 6.4 and the GM value decreased with decreasing pH from 6.4 to 5.4. This was due to an excessive loss of INS Ca from CM. There was a decrease in GM with the increase in the concentration of added EDTA, which was probably due to the loss of colloidal calcium phosphate, which weakens the integrity of CM. Loss tangent (LT) values at GM increased with a reduction in milk pH and the addition of EDTA to milk. Rennet gels at the point of the GM were subjected to constant low shearing to fracture the gels. With a reduction in INS Ca content, the yield stress decreased, whereas LT values increased indicating a weaker, more flexible casein network. Microstructure of rennet-induced gels near the GM point and 2 to 10 h after this point was studied using fluorescence microscopy. At GM, gels made from milk acidified to pH 6.4 exhibited more branched, interconnected networks, whereas strands and clusters became larger with a reduction in milk pH to 5.4. Gels made from milk with EDTA added had more finely dispersed protein clusters compared with gels made from milk with no EDTA added. These microscopic observations supported the effect of loss of INS Ca on GM and LT. There was a decrease in apparent interconnectivity between strands in gel microstructure during aging, which agreed with the decrease in G′ after GM. It can be concluded that low levels of solubilization of INS Ca and the decrease in milk pH resulted in an increase in GM. With greater losses of INS Ca there was excessive reduction in cross-linking within CM, which resulted in weaker, more flexible rennet gels. This complex behavior cannot be explained by adhesive hard sphere models for CM or rennet gels made from these CM.     

Clotting and proteolytic properties of plant coagulants in regular and ultrafiltered bovine skim milk

Regular and ultrafiltered (UF; 1×, 2× and 4× concentrated) skim milk samples were treated with a range of enzymes including calf rennet, ficin and papain. The clotting properties, curd casein profiles and free amino acid (FAA) contents were determined. In general, UF milk samples coagulated faster and formed firmer curds irrespective of protein concentration. Furthermore, both ficin and papain had a more significant effect on proteolysis in curd formed from regular and 1× UF milk than on 2× or 4× UF milk. Cardoon extract and calf rennet had very similar clotting properties, although the former caused both the capillary electrophoresis profile of caseins and FAA measurements to show slightly more extensive hydrolysis in the curd. The results suggest that the UF process may cause structural changes to proteins or other milk constituents with a resultant change in clotting properties and proteolysis of the casein molecules.     

Effect of pH at heat treatment on the hydrolysis of κ-casein and the gelation of skim milk by chymosin

Skim milk was adjusted to pH values between 6.5 and 7.1 and heated at 90 °C for times from 0 to 30 min. After heat treatment, the samples were re-adjusted to the natural pH (pH 6.67) and allowed to re-equilibrate. High levels of denatured whey proteins associated with the casein micelles during heating at pH 6.5 (about 70-80% of the total after 30 min of heating). This level decreased as the pH at heating was increased, so that about 30%, 20% and 10% of the denatured whey protein was associated with the casein micelles after 30 min of heating at pH 6.7, 6.9 and 7.1, respectively. Increasing levels of κ-casein were transferred to the serum as the pH at heating was increased. The loss of κ-casein and the formation of para-κ-casein with time as a consequence of the chymosin treatment of the milk samples were monitored by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The loss of κ-casein and the formation of para-κ-casein were similar for the unheated and heated samples, regardless of the pH at heating or the heat treatment applied. Monitoring the gelation properties with time for the chymosin-treated milk samples indicated that the heat treatment of the milk markedly increased the gelation time and decreased the firmness (G^′) of the gels formed, regardless of whether the denatured whey proteins were associated with the casein micelles or in the milk serum. There was no effect of pH at heat treatment. These results suggest that the heat treatment of milk has only a small effect on the primary stage of the chymosin reaction (enzymatic phase). However, heat treatment has a marked effect on the secondary stage of this reaction (aggregation phase), and the effect is similar regardless of whether the denatured whey proteins are associated with the casein micelles or in the milk serum as nonsedimentable aggregates.     

Acid induced gelation behavior of skim milk concentrated by membrane filtration

This work reports a detailed study of the effect of ultrafiltration (UF) and diafiltration (DF) on the acid-induced gelation behavior of fresh milk retentates (2× and 4×). Concentrates were heated at 80°C for 15 min, and compared to unheated samples. The use of extensive DF caused a significantly greater amount of protein (both caseins and whey proteins) in the supernatant fraction, compared to UF retentates at the same concentration, both in unheated and heated samples. DF retentates showed higher pH of gelation, compared to the corresponding UF retentates. The development of tan δ is reported for the first time as a function of colloidal calcium release, and the protein gelation behavior discussed in light differences in composition of the soluble fraction. The results demonstrate how processing history can affect compositional changes and the gelation behavior of fresh milk retentates. Membrane filtration is a widespread unit operation in the dairy industry, employed either to prepare fresh concentrates for further processing, or ingredients with specific functional properties. This work describes in detail the effect of processing history during membrane filtration on the rheological properties of acid induced gels and will help in optimizing formulations and prepare the right ingredients for the right application. It will also be possible to determine new ways to define processing quality of the milk protein concentrates, as it relates to their ability to form texture in fermented dairy products.     

Effect of pH at heating on the acid-induced aggregation of casein micelles in reconstituted skim milk

Reconstituted skim milk samples at pH between 6.5 and 7.1 (heating pH) were heated at 80°C, 90°C or 100°C for 30 min (heating temperature). The particle size of the casein micelles was measured at pH 4.75-7.1 (measurement pH) and at temperatures of 10°C, 20°C and 30°C (measurement temperature) using photon correlation spectroscopy. The particle size of the casein micelles, at a measurement pH of 6.7 and a measurement temperature of 20°C, was dependent on the heating pH and heating temperature to which the milk was subjected. The casein micelle size in unheated milk was about 215 nm. At a heating pH of 6.5, the casein micelle size increased by about 15, 30 and 40 nm when the milk was heated at 80°C, 90°C or 100°C, respectively. As the heating pH of the milk was increased, the size of the casein micelles decreased so that, at pH 7.1, the casein micelles were ∼20 nm smaller than those from unheated milk. Larger effects were observed as the heating temperature was increased from 80°C to 100°C. The size differences as a consequence of the heating pH were maintained at all measurement temperatures and at all measurement pH down to the pH at which aggregation of the micelles was observed. For all samples, size measurements at 10°C showed no aggregation at all measurement pH. Aggregation occurred at progressively higher pH as the measurement temperature was increased. Aggregation also occurred at a progressively higher measurement pH as the heating pH was increased. The particle size changes on heating and the aggregation on subsequent acidification may be related to the pH dependence of the association of whey proteins with, and the dissociation of κ-casein from the casein micelles as milk is heated.     

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.     

Gelation of casein micelles in ��-casein reduced milk prepared using membrane filtration

Pasteurized skim milk was subjected to membrane filtration using a molecular weight cut-off of 80 kDa and a plate and frame pilot scale system at temperatures below 10 °C. Via this process, transmission of whey proteins and ??-casein through the membrane was achieved. The milk was concentrated to two times (based on volume reduction), and whey protein-free permeate was added to return to the original volume fraction of casein micelles in milk. This diafiltration process was carried out four times, and the retentate obtained was nearly free of whey proteins and with approximately 20% of ??-casein removed. The same membrane filtration was also carried out at 25 °C to achieve transmission of whey protein but not of ??-casein, and to obtain whey protein-depleted milk without depletion of ??-casein.The gelling behaviour of these samples, reconstituted to the original casein volume fraction, was examined using rheology and diffusing wave spectroscopy. When compared to the original skim milk it was found that there were no statistically significant differences in gelation behaviour during acidification, but differences were noted in gelation time and final stiffness modulus for samples undergoing renneting. These differences were attributed mostly to the changes in ionic composition, as when the serum composition of the retentates was re-equilibrated against the original skim milk by dialysis; the gelation behaviour of the samples was comparable to that of skim milk. The results clearly indicate the importance of the milk's overall ionic balance in the early stages of aggregation of rennet-induced gelation of milk.     

Standardization of milk using cold ultrafiltration retentates for the manufacture of parmesan cheese

The effects of using cold ultrafiltered (UF) retentates (both whole and skim milk) on the coagulation, yield, composition, and ripening of Parmesan cheese were investigated. Milks for cheese making were made by blending cold UF retentates with partially skimmed milk to obtain blends with 14.2% solids and a casein:fat ratio of 1.1. Cutting times, as selected by the cheese-maker, were approximately 15 and approximately 20 min for experimental and control milks, respectively. Storage modulus values at cutting were similar, but yield stress values were significantly higher in UF retentate standardized milks. Cheese yields were significantly higher in UF retentate standardized milks (approximately 12%) compared with control milk (cream removed) (approximately 7 to 8%). Significantly higher protein recoveries were obtained in cheeses manufactured using cold UF retentates. There were no differences in the pH and moisture contents of the cheeses prior to brining, and there was no residual lactose or galactose left in the cheeses. Using UF retentates resulted in a significant reduction in whey volume as well as a higher proportion of protein in the solids of the whey. Proteolysis, free fatty acids, and sensory properties of the cheeses were similar. The use of milk concentrated by cold UF is a promising way of improving the yield of Parmesan cheese without compromising cheese quality. The question remaining to be answered by the cheesemaker is whether it is economical to do so.     

Impact of ultrafiltration/diafiltration sequence on the production of soy protein isolate by membrane technologies

In previous studies, the advantages of combining electrodialysis using a bipolar-cationic membranes configuration to acidify a soy protein extract to pH 6 with ultrafiltration/diafiltration (UF/DF) using a 100 kDa membrane to produce a soy protein isolate with low phytic acid content and improved solubility between pH 2 and 4 was demonstrated, when compared to the production of soy protein isolates by isoelectric precipitation and by UF/DF of a soy protein extract at pH 9. However, limited work was done to establish the impact of the UF/DF sequence for the purification of the pH 6 extract. Therefore, the purpose of this work was to study the impact of four different UF/DF sequences with a total permeate volume of 1.5–1.6 times the initial volume, on membrane fouling and permeate flux, as well as on the isolate protein, ash and phytic acid contents and solubility profile. Of the investigated UF/DF sequences, the VCR 5, VD 4 sequence was shown to be the one with the most severe fouling and consequently the most severe permeate flux decline. At the same time, it was also the VCR 5, VD 4 sequence which was the most efficient in terms of ash and phytic acid removal, followed by the VCR 5, re-VCR5 sequence, the VCR 2, VD 2 sequence and the VCR 2, (re-VCR 2)X 2 sequence, respectively. It was also observed that isolate with low phytic acid content resulted in narrower protein solubility profiles around the isoelectric point and higher protein solubility for the pH range of 2 to 4.Industrial relevancePlant proteins have made up a higher proportion of the human diet in recent years. Soybeans are the most important source of plant protein ingredients accounting for some 68% of global plant protein consumption in the world. Soy protein isolate is traditionally prepared by isoelectric precipitation process. This process has high productivity, however, it results in products with poor functional properties due to protein denaturation and to the presence of phytic acid (1–3% w/w) which alters the solubility of the isolates especially for the pH below the proteins' isoelectric point. In this work, we combined electrodialysis using a bipolar-cationic membranes configuration to acidify a soy protein extract to pH 6 with ultrafiltration/diafiltration (UF/DF) using a 100 kDa membrane to produce soy protein isolates with low phytic acid content. The impact of four different UF/DF sequences on membrane fouling, permeate flux, isolate composition and solubility profile was studied. Of the investigated UF/DF sequences, the VCR 5, VD 4 sequence was shown to be the one with the most severe fouling but at the same time the most efficient in terms of ash and phytic acid removal. It was also observed that the isolate produced by the VCR 5, VD 4 sequence shows narrower protein solubility profiles around the isoelectric point and higher protein solubility for the pH range of 2 to 4 than isolates produced by alternative UF/DF sequences. This isolate could be considered as a valuable ingredient for the formulation of fruit juice beverages or power juices, considering that the pH of these liquid food products is around 3.5.