Effects of milk heat treatment and solvent composition on physicochemical and selected functional characteristics of milk protein concentrate

Milk protein concentrate (MPC) powders (～81% protein) were made from skim milk that was heat treated at 72°C for 15 s (LHMPC) or 85°C for 30 s (MHMPC). The MPC powder was manufactured by ultrafiltration and diafiltration of skim milk at 50°C followed by spray drying. The MPC dispersions (4.02% true protein) were prepared by reconstituting the LHMPC and MHMPC powders in distilled water (LHMPC_w and MHMPC_w, respectively) or milk permeate (LHMPC_p and MHMPC_p, respectively). Increasing milk heat treatment increased the level of whey protein denaturation (from ～5 to 47% of total whey protein) and reduced the concentrations of serum protein, serum calcium, and ionic calcium. These changes were paralleled by impaired rennet-induced coagulability of the MHMPC_w and MHMPC_p dispersions and a reduction in the pH of maximum heat stability of MHMPC_p from pH 6.9 to 6.8. For both the LHMPC and MHMPC dispersions, the use of permeate instead of water enhanced ethanol stability at pH 6.6 to 7.0, impaired rennet gelation, and changed the heat coagulation time and pH profile from type A to type B. Increasing the severity of milk heat treatment during MPC manufacture and the use of permeate instead of water led to significant reductions in the viscosity of stirred yogurt prepared by starter-induced acidification of the MPC dispersions. The current study clearly highlights how the functionality of protein dispersions prepared by reconstitution of high-protein MPC powders may be modulated by the heat treatment of the skim milk during manufacture of the MPC and the composition of the solvent used for reconstitution.     

Effects of polymerized goat milk whey protein on physicochemical properties and microstructure of recombined goat milk yogurt

Goat milk whey protein concentrates were manufactured by microfiltration (MF) and ultrafiltration (UF). When MF retentate blended with cream, which could be used as a starting material in yogurt making. The objective of this study was to prepare goat milk whey protein concentrates by membrane separation technology and to investigate the effects of polymerized goat milk whey protein (PGWP) on the physicochemical properties and microstructure of recombined goat milk yogurt. A 3-stage MF study was conducted to separate whey protein from casein in skim milk with 0.1-µm ceramic membrane. The MF permeate was ultrafiltered using a 10 kDa cut-off membrane to 10-fold, followed by 3 step diafiltration. The ultrafiltration-diafiltration-treated whey was electrodialyzed to remove 85% of salt, and to obtain goat milk whey protein concentrates with 80.99% protein content (wt/wt, dry basis). Recombined goat milk yogurt was prepared by mixing cream and MF retentate, and PGWP was used as main thickening agent. Compared with the recombined goat milk yogurt without PGWP, the yogurt with 0.50% PGWP had desirable viscosity and low level of syneresis. There was no significant difference in chemical composition and pH between the recombined goat milk yogurt with PGWP and control (without PGWP). Viscosity of all the yogurt samples decreased during the study. There was a slight but not significant decrease in pH during storage. Bifidobacterium and Lactobacillus acidophilus in yogurt samples remained above 10⁶ cfu/g during 8-wk storage. Scanning electron microscopy of the recombined goat milk yogurt with PGWP displayed a compact protein network. Results indicated that PGWP prepared directly from raw milk may be a novel protein-based thickening agent for authentic goat milk yogurt making.     

Gelation of Mixed Gels Containing κ-Carrageenan and Skim Milk Components

ABSTRACT: The gelation characteristics of mixed gels containing κ-carrageenan and skim milk or milk fractions (skim milk permeate or retentate) obtained by ultrafiltration were examined. Increasing the skim milk solids content of mixtures containing carrageenan increased setting temperatures and gel strength. The milk proteins contributed to gel strength but did not influence the setting temperature of mixtures. The binding of denatured whey proteins to casein micelles affected gel network formation of milk-carrageenan mixtures containing 10% milk solids. Network formation in mixed gels containing carrageenan and milk or milk fractions was initiated by the carrageenan component and dictated primarily by the ionic content of the mixtures.     

Influence of partially demineralized milk proteins on rheological properties and microstructure of acid gels

Innovative clean label processes employed in the manufacture of acid gels are targeted to modify the structure of proteins that contribute to rheological properties. In the present study, CO₂-treated milk protein concentrate powder with 80% protein in dry matter (TMPC80) was mixed with nonfat dry milk (NDM) in different ratios for the manufacture of acid gels. Dispersions of NDM and TMPC80 that provided 100, 90, 70, and 40% of protein from NDM were reconstituted to 4.0% (wt/wt) protein and 12.0% (wt/wt) total solids. Dispersions were adjusted to pH 6.5, followed by heat treatment at 90°C for 10 min. Glucono-δ-lactone was added and samples were incubated at 30°C, reaching pH 4.5 ± 0.05 after 4 h of incubation. Glucono-δ-lactone levels were adjusted to compensate for the lower buffering capacity of samples with higher proportions of TMPC80, which is attributable to the depletion of buffering minerals from both the serum and micellar phase during preparation of TMPC80. Sodium dodecyl sulfate-PAGE analysis indicated a higher amount of caseins in the supernatant of unheated suspensions with increasing proportions of CO₂-treated TMPC80, attributable to the partial disruption of casein micelles in TMPC80. Heat treatment reduced the level of whey proteins in the supernatant due to the heat-induced association of whey proteins with casein micelles, the extent of which was larger in samples containing more micellar casein (i.e., samples with a lower proportion of TMPC80). Particle size analysis showed only small differences between nonheated and heated dispersions. Gelation pH increased from ?5.1 to ?5.3, and the storage modulus of the gels at pH 4.5 increased from ?300 to ?420 Pa when the proportion of protein contributed by TMPC80 increased from 0 to 60%. Water-holding capacity also increased and gel porosity decreased with increasing proportion of protein contributed by TMPC80. The observed gel properties were in line with microstructural observations by confocal microscopy, wherein sample gels containing increasing levels of TMPC80 exhibited smaller, well-connected aggregates with uniform, homogeneous pore sizes. We concluded that TMPC80 can be used to partially replace NDM as a protein source to improve rheological and water-holding properties in acid gels. The resultant gels also exhibited decreased buffering, which can improve the productive capacity of yogurt manufacturing plants. Overall, the process can be leveraged to reduce the amount of hydrocolloids added to improve yogurt consistency and water-holding capacity, thus providing a path to meet consumer expectations of clean label products.     

Power ultrasound as a tool to improve the processability of protein-enriched fermented milk gels for Greek yogurt manufacture

One approach to avoid production of acid whey during the manufacture of high-protein yogurt and related products is to concentrate the milk before fermentation. However, the resultant gels are firm so that stirring in the tank and further processing are difficult on an industrial scale. We hypothesize that power ultrasound (US) during fermentation softens the gel because sound waves cause cavitation and strong shear forces in the fluid. Skim milk was standardized to different protein contents up to 12%, heated (85°C, 30 min), and acidified with thermophilic or mesophilic starter cultures. An excessive increase in gel firmness as a function of protein content was detected. In the next series of experiments, US was applied during fermentation. Milks (10% protein) were acidified at 43.5°C and sonicated from pH 5.8 to 5.1 with a sonotrode (20 kHz, 20 W). Immediately after fermentation, gels were agitated using a rheometer with a vane geometry. The maximum torque required to break the gel was reduced by 75% following US, and gel firmness was reduced by 80%. Gels were then processed into stirred yogurt and analyzed. Sonicated samples were smoother with fewer large aggregates. Confocal laser scanning microscopy images suggested a less cohesive structure and more compact microgel particles, resulting in reduced viscosity. We concluded that US is a promising tool to weaken the gel and facilitate further processing. This enables new approaches for the manufacture of Greek yogurt, particularly in regard to avoiding production of acid whey and developing products with novel textures.     

Pre-treatment methods of Edam cheese milk. Effect on the whey composition

Edam cheese milk was subjected to high-heat treatment (HH), ultrafiltration (UF) and microfiltration (MF). The effect on the recovery yield and the composition of whey was studied. Traditional Edam process was used as a reference. HH reduced the whey protein concentration of milk and whey, but the recovery from milk to whey was not affected. Reduction of whey proteins was the highest (28%) with MF treatment, during which 15% was lost in the MF permeate and 13% was co-precipitated with the cheese curd. Co-precipitation of the whey proteins was the highest (84%) with ultrafiltered milk. MF and UF treatments produced 22% less whey with increased whey protein concentration. Elevation of the cheese milk protein concentration by microfiltration or ultrafiltration decreased the recovery of fat in whey. None of the treatments decreased the residual casein concentration in whey. The protein composition was altered by UF and MF treatments, which significantly increased the caseinomacropeptide content of total protein in whey. The whey was processed into whey protein concentrate powders. The amino acid composition of the whey protein concentrate produced from microfiltration process was significantly different from the others.     

Manufacture of high-protein yogurt without generating acid whey – Impact of the final pH and the application of power ultrasound on texture properties

The fermentation of preconcentrated milk is a challenging method to avoid acid whey during the manufacture of high-protein fermented milks like Greek yogurt. Milk concentrates (10% protein) were fermented to a final pH of 5.0, 4.8, or 4.6 and processed into stirred yogurt. Additionally, the potential of power ultrasound (US) as a post-processing tool was examined by sonicating the stirred yogurt with a sonotrode at 20 kHz. Set gels fermented to pH 4.8 and 5.0 were considerably softer than gels fermented to pH 4.6. Stirred yogurts fermented to pH 4.8 or 5.0 were less grainy and exhibited a reduced apparent viscosity and water-holding capacity. The application of US further decreased the visual graininess and product viscosity whereas the particle size was only slightly affected. The final pH and sonication are two powerful approaches to control the rheological properties of high-protein fermented milks, offering the potential for innovative processes and products.     

Effect of skim milk treated with high hydrostatic pressure on permeate flux and fouling during ultrafiltration

Ultrafiltration (UF) is largely used in the dairy industry to generate milk and whey protein concentrate for standardization of milk or production of dairy ingredients. Recently, it was demonstrated that high hydrostatic pressure (HHP) extended the shelf life of milk and improved rennet coagulation and cheese yield. Pressurization also modified casein micelle size distribution and promoted aggregation of whey proteins. These changes are likely to affect UF performance. Consequently, this study determined the effect of skim milk pressurization (300 and 600 MPa, 5 min) on UF performance in terms of permeate flux decline and fouling. The effect of HHP on milk proteins was first studied and UF was performed in total recycle mode at different transmembrane pressures to determine optimal UF operational parameters and to evaluate the effect of pressurization on critical and limiting fluxes. Ultrafiltration was also performed in concentration mode at a transmembrane pressure of 345 kPa for 130 or 140 min to evaluate the decline of permeate flux and to determine fouling resistances. It was observed that average casein micelle size decreased by 32 and 38%, whereas β-lactoglobulin denaturation reached 30 and 70% at 300 and 600 MPa, respectively. These results were directly related to UF performance because initial permeate fluxes in total recycle mode decreased by 25% at 300 and 600 MPa compared with nonpressurized milk, critical flux, and limiting flux, which were lower during UF of milk treated with HHP. During UF in concentration mode, initial permeate fluxes were 30% lower at 300 and 600 MPa compared with the control, but the total flux decline was higher for nonpressurized milk (62%) compared with pressure-treated milk (30%). Fouling resistances were similar, whatever the treatment, except at 600 MPa where irreversible fouling was higher. Characterization of the fouling layer showed that caseins and β-lactoglobulin were mainly involved in membrane fouling after UF of pressure-treated milk. Our results demonstrate that HHP treatment of skim milk drastically decreased UF performance.     

Acid-induced gelation of whey protein polymers: effects of pH and calcium concentration during polymerization

Heating whey protein dispersions (90°C for 15 min) at low ionic strength and pH values far from isoelectric point (pH>6.5) induced the formation of soluble polymers. The effect of mineral environment during heating on the hydrodynamic characteristics and acid-induced gelation properties of polymers was studied. Whey protein dispersions (80 g/l) were denatured at different pH (6.5–8.5) and calcium concentrations (0–4 mm) according to a factorial design. At pH 6.5, the hydrodynamic radius of protein polymers increased with increasing calcium concentration, while the opposite trend was observed at pH 8.5. Intrinsic viscosity results suggested that heating conditions altered the shape of protein polymers. Whey protein polymers were acidified to pH 4.6 with glucono-δ-lactone and formed opaque particulate gels. The storage modulus and firmness of gels were both affected by conditions used to prepare protein polymers. As a general trend, polymers with high intrinsic viscosity produced stronger gels, suggesting a relationship between polymer shape and gel strength.Acid gelation properties of whey protein polymers makes them suitable ingredients for yoghurt applications. Using whey protein polymers to standardize protein content increased yoghurt viscosity to 813 Pa.s while using skim milk powder at same protein concentration increased yoghurt viscosity to 393 Pa.s. Water holding capacity of protein polymers in yoghurt was 19.8 ml/g compared to 7.2 ml/g for skim milk powder protein. Acid gelation properties of whey protein polymers are modulated by calcium concentration and heating pH and offers new alternatives to control the texture of fermented dairy products.     

Front-face fluorescence spectroscopy combined with chemometrics to detect high proteinaceous matter in milk and whey ultrafiltration permeate

Proteinaceous matter can leak into the permeate stream during ultrafiltration (UF) of milk and whey and lead to financial losses. Although manufacturers can measure protein content in the finished permeate powders, there is currently no rapid monitoring tool during UF to identify protein leak. This study applied front-face fluorescence spectroscopy (FFFS) and chemometrics to identify the fluorophore of interest associated with the protein leak, develop predictive models to quantify true protein content, and classify the types of protein leak in permeate streams. Crude protein (CP), nonprotein nitrogen (NPN), true protein (TP), tryptone-equivalent peptide (TEP), α-lactalbumin (α-LA), and β-lactoglobulin (β-LG) contents were measured for 37 lots of whey permeate and 29 lots of milk permeate from commercial manufacturers. Whey permeate contained more TEP than did milk permeate, whereas milk permeate contained more α-LA and β-LG than did whey permeate. The types of protein leak were thus identified for predictive model development. Based on excitation-emission matrix (EEM) of high- and low-TP permeates, tryptophan excitation spectra were collected for predictive model development, measuring TP content in permeate. With external validation, a useful model for quality control purposes was developed, with a root mean square error of prediction of 0.22% (dry basis) and a residual prediction deviation of 2.8. Moreover, classification models were developed using partial least square discriminant analysis. These classification methods can detect high TP level, high TEP level, and presence of α-LA or β-LG with 83.3%, 84.8%, and 98.5% cross-validated accuracy, respectively. This method showed that FFFS and chemometrics can rapidly detect protein leaks and identify the types of protein leak in UF permeate. Implementation of this method in UF processing plants can reduce financial loss from protein leaks and maintain high-quality permeate production.     

Effect of pressure treatment of recombined whole milk on fat globule membrane composition and acid gelation functionality

Recombined whole milk was prepared by pressure treating skim milk (200 to 600 MPa/30 min) then homogenizing with milkfat (HPHO) or by homogenizing milkfat with skim milk then pressure treating the recombined whole milk (HOHP). β-Lactoglobulin denaturation increased at higher pressures. Low levels of α-lactalbumin were denatured at 600 MPa only. Denaturation was similar in the HPHO and HOHP milk. The HPHO milk had statistically similar levels of total protein, higher levels of whey protein and κ-casein and lower levels of α_s-casein adsorbed to the fat globules compared with the HOHP milk. The HPHO milk had a higher proportion of β-casein directly at the interface at all pressures and a higher proportion of κ-casein and a lower proportion of denatured whey proteins at pressure up to 400 MPa than the HOHP milk. Acid gels prepared from the HOHP milk had higher final G′ and yield stresses than those from the HPHO milk. These differences are discussed in relation to the compositions of the proteins adsorbed to the fat globules and how these interact during acidification.     

Fermentation of specially formulated milk with single strains of bifidobacteria

A number of strains of bifidobacteria have been used to ferment milk to a yoghurt-like product. The milk used for fermentation was fortified with whey protein and threonine. It was prepared by mixing skimmed milk and cheddar cheese whey which had been concentrated using ultrafiltration membranes which allowed lactose to permeate. A 2 percent inoculum was used, and fermentation was carried out at 37° C overnight. The resulting products resembled voghurt, having good consistency. 'walnutty' aroma (acetaldehyde). and pleasant acidity.     

Thermal Aggregation of Whey Proteins in Model Solutions as Affected by Casein/Whey Protein Ratios

Model solutions (32.5 g protein/L) prepared from milk, ultrafiltration permeate, and whey protein isolate were adjusted at pH 6.7 to casein:whey protein (C:W) ratios of 80:20, 60:40, 40:60,20:80, and 0:100. Heating was performed in test tubes at 95 °C for 5 min. Observations of the heated suspensions revealed the occurrence of heterogeneous particulates from the existing casein micelles complexed with denatured whey proteins and from aggregates essentially consisting of denatured whey proteins. The proportion of whey protein aggregates increased as C:W was changed from 80:20 to 20:80. The results from this study confirmed that heat-induced aggregates were formed not only from casein micelles but also from heat-denatured whey proteins.     

High pressure-induced changes in milk proteins and possible applications in dairy technology

This paper reviews the newest information on the effects of high pressure (HP) on whey proteins, caseins and milk enzymes, and discusses their influence on milk properties. HP treatments cause substantial modification to milk proteins and to the mineral balance of milk. Casein micelles disaggregate into smaller particles or aggregate, depending the intensity and the temperature of the HP treatment. Whey proteins are denatured, possibly interacting with the remnants of the casein micelles, and give aggregated forms different from those produced by heat treatment. These events influence rennet coagulation properties of milk, with micellar disintegration favoring coagulation and whey protein denaturation hindering the aggregation of renneted micelles and enhancing cheese yield. HP treatment of milk favors acid coagulation and produces acid gels whose structure is greatly determined by the different micellar sizes attainable and the degree of whey protein denaturation. Milk gels can also be formed from concentrated milk under HP, providing new structures inaccessible via conventional methods.     

Preliminary studies on the gelation processes of fermented and GDL-acidified bovine and caprine milk systems

Bovine and caprine milk gels were made by GDL-acidification and yogurt fermentation (using 'ropy' and 'non-ropy' starter cultures). The respective gelation processes were monitored rheologically using dynamic oscillatory testing. The bovine fermented systems produced gel structures with about half the strength of the equivalent chemically acidified gels. The fermented caprine milk systems produced gel structures some eight to 10 times weaker than the equivalent acidified systems. In all cases the caprine systems were weaker than the bovine gels despite having higher protein contents. In all cases the 'ropy' milk systems followed somewhat different gelation patterns and formed weaker gels than the equivalent 'non-ropy' and GDL-acidified systems. These data suggested that the starter culture material (biomass and extracellular polysaccharides) may have interfered with the protein–protein interactions during yogurt fermentation. This produced weaker gel structures, possibly by a modified gelation mechanism .     

Transglutaminase cross-linking of milk proteins and impact on yoghurt gel properties

A novel yoghurt process was investigated in which milk proteins were covalently cross-linked by a microbial transglutaminase (TG) preparation containing glutathione (TG+GSH). As unheated milk is normally less reactive towards TG, TG+GSH was applied to enable non-inhibited cross-linking without requiring a pre-heat treatment beyond pasteurisation conditions. After the TG+GSH incubation phase, the enzyme was inactivated by heat treatment of the yoghurt milk prior to fermentation. During yoghurt fermentation, no negative effect of TG+GSH on fermentation time was found. Protein cross-linking by TG+GSH was enhanced, resulting in higher apparent viscosity and a higher degree of protein polymerisation than that given by TG without GSH. Furthermore, different ratios of casein to whey proteins (CWP ratios) were used to investigate the effect of both protein fractions on covalently cross-linked yoghurt gel structures. The results showed a relationship between extent of cross-linking, apparent viscosity and CWP ratio of the yoghurt gels. During storage for up to 6 weeks at 4 °C, no changes in rheological properties and degree of protein polymerisation were measurable for stirred yoghurt gels prepared from cross-linked milk proteins.     

Gelation of casein-whey protein mixtures

Heated milk consists of a mixture of whey protein-coated casein micelles and soluble whey protein aggregates. The acid-induced gelation properties of heated milk are consistently different from those of unheated milk—i.e., a shift in gelation pH, stronger gels, and a different microstructure of the gels. In this study we investigated the role of the different fractions of denatured whey proteins on the acid-induced gelation, the gel hardness, and the microstructure. Both whey protein fractions contribute to the observed shift in gelation pH, although by a different mechanism. Obtaining gels with high gel hardness occurs most effectively when all denatured whey proteins are present as whey protein aggregates. It was observed that disulfide bridge exchange reactions during the acid-induced gelation at ambient temperature play an important role for both whey protein fractions. Additionally, disulfide interactions seem to occur between the aggregates and the casein micelles during the gel state. In this study, we show the development of a new approach for confocal scanning laser microscopy measurements—i.e., separate staining of the proteins in milk. By using this method, we were able to determine that, although whey protein aggregates are not linked to the casein micelles, they nevertheless gel at the same moment. This work adds to a better understanding of the role of denatured whey proteins during acid-induced gelation and could improve the effective use of whey proteins.     

High-pressure treatment of milk in industrial and pilot-scale equipments: effect of the treatment conditions on the protein distribution in different milk fractions

The effects of two different high-pressure (HP) equipments, operating at industrial- and pilot scales, and of the HP-release rate on the contents of non-sedimentable proteins and denatured whey proteins were investigated after treatments of skim milk—from 250 to 650 MPa. Non-sedimentable caseins and denatured whey proteins significantly increased with the pressure level. The industrial-scale equipment produced lower micellar disintegration than the pilot-scale equipment with similar degrees of whey protein denaturation. Ultracentrifugation supernatants obtained from skim milk at 100,000×g and 20 °C for 1 h were also HP-treated for comparative purposes, showing that, in skim milk, the presence of casein promoted the denaturation of whey proteins, although the extent of whey protein denaturation did not influence the release of casein to the soluble phase. Furthermore, most denatured whey proteins remained soluble after treatment in both equipments. In the pilot-scale equipment, the pressure-release rate influenced casein solubilization and whey protein denaturation.     

乳成分对酸奶凝胶微观结构和流变学性质的影响

酸奶凝胶的许多宏观的物理特性与其微观结构和流变学性质密切相关。从酸奶的微结构、流变学性质和质地等方面综述了乳脂肪、蛋白质及调节酪蛋白和乳清蛋白比例对酸奶凝胶的影响。     

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.