Effects of milk pH alteration on casein micelle size and gelation properties of milk

The effects of changes in pH above and below the natural pH of milk (ca 6.6) on the casein micelle size and the gelation properties of the pH adjusted and restored samples were investigated. The size of casein micelles increased at pH 7.0 and 7.5, then started to decrease above pH 8.5. It is postulated that at pH below 8.5 the casein micelles swell, while elevated pH causes their dissociation. Conversely, the size of casein micelles decreased on acidification to pH 5.5 and increased when the pH dropped below 5.5, due to the shrinkage of casein micelles at lower pH before their aggregation at pH below 5.5. In response to neutralising treated milk back to normal milk pH of 6.6, it was found that the casein micelle size of treated milk with a narrow range of pH change between 6.0 and 7.0 was reversible, while beyond this range the structure of casein micelles became irreversible. The restoration of casein micelle size was followed by the restoration of some parameters such as soluble calcium, ethanol stability, and milk whiteness. Acid-induced gelation did not change the elastic modulus, while rennet-induced gelation was affected by initial milk pH. In reference to the size of reversible range elastic modulus (G’) of acid or rennet gels made from restored milk, the sizes were similar to control milk after 6 h of gelation.     

Importance of casein micelle size and milk composition for milk gelation

The economic output of the dairy industry is to a great extent dependent on the processing of milk into other milk-based products such as cheese. The yield and quality of cheese are dependent on both the composition and technological properties of milk. The objective of this study was to evaluate the importance and effects of casein (CN) micelle size and milk composition on milk gelation characteristics in order to evaluate the possibilities for enhancing gelation properties through breeding. Milk was collected on 4 sampling occasions at the farm level in winter and summer from dairy cows with high genetic merit, classified as elite dairy cows, of the Swedish Red and Swedish Holstein breeds. Comparisons were made with milk from a Swedish Red herd, a Swedish Holstein herd, and a Swedish dairy processor. Properties of CN micelles, such as their native and rennet-induced CN micelle size and their ζ-potential, were analyzed by photon correlation spectroscopy, and rennet-induced gelation characteristics, including gel strength, gelation time, and frequency sweeps, were determined. Milk parameters of the protein, lipid, and carbohydrate profiles as well as minerals were used to obtain correlations with native CN micelle size and gelation characteristics. Milk pH and protein, CN, and lactose contents were found to affect milk gelation. Smaller native CN micelles were shown to form stronger gels when poorly coagulating milk was excluded from the correlation analysis. In addition, milk pH correlated positively, whereas Mg and K correlated negatively with native CN micellar size. The milk from the elite dairy cows was shown to have good gelation characteristics. Furthermore, genetic progress in relation to CN micelle size was found for these cows as a correlated response to selection for the Swedish breeding objective if optimizing for milk gelation characteristics. The results indicate that selection for smaller native CN micelles and lower milk pH through breeding would enhance gelation properties and may thus improve the initial step in the processing of cheese.     

Influence of variation in calcium content on casein micelle stability and techno-functional properties of buffalo milk

Changes in techno-functional properties of buffalo milk were evaluated due to variation in calcium content. Decalcification resulted in significant variation in ζ-potential, casein size, colour and η_app. However, calcium addition only influenced ζ-potential of milk. In case of acid gelation, the time and temperature required for coagulation decreased significantly for both calcium-depleted and -added milks. However, during chymosin gelation, only 20%–30% of calcium-depleted milk coagulated with an increased clotting time. Furthermore, calcium addition increased firmness, consistency and cohesiveness of both chymosin and acid-induced gelation.     

Natural variation of bovine milk fat globule size within a herd

Native milk fat globule (MFG) size, pH, total fat, protein, and lactose composition in milk from individual cows was examined in autumn and spring 2012 and autumn 2013. Mean MFG diameters ranged between 2.5 and 5.7 µm. Some cows were observed to consistently produce small or large MFG throughout the sampling period, though trends were not consistent across all cows. Small-MFG milk contained more total polar lipids; however, the relative proportion of individual polar lipids did not differ with MFG size, with the exception of phosphatidylcholine, which was greater in small-MFG milk. No significant correlation between MFG size and proximate composition, including calculated fat yield or pH, was found. This work revealed a natural cow-to-cow variation in MFG size exists, which could potentially be exploited to improve the functionality of milk for manufacturing and some products.     

Native milk fat globule size and its influence on whipping properties

The objective of this study was to investigate the influence of native milk fat globule size on the aeration of high fat dairy products with regard to maximum firmness time, gas inclusion and foam stability. The results showed that whipping time to maximum firmness was inversely proportional to mean fat globule size for both unhomogenised and slightly homogenised (2 MPa) creams. Additionally, increasing native mean fat globule size of the creams resulted in increased overrun. No significant differences in serum drainage were found between creams with different native milk fat globule size. Furthermore, when creams with native large mean fat globules were homogenised, the results showed that the maximum firmness time was in accordance with the mean fat globule size of non-aggregated creams. In the present study, cream fractionation was achieved by creaming or in a cost effective and fast manner using a modified centrifugal separator.     

Heat and/or high-pressure treatment of skim milk: changes to the casein micelle size, whey proteins and the acid gelation properties of the milk

Skim milk was subjected to heat, pressure or combined processes. In general, higher levels of whey protein denaturation were observed for milk subjected to combined processes than those heat- or pressure-treated only. Heat treatment caused small changes to the casein micelle size. Pressure treatment decreased the casein micelle size; however, the effect was less marked when heat and pressure treatments were combined. Acidification of the skim milks produced gels with a range of firmness, yield stresses and yield strains depending on the treatments applied. These changes in acid gel properties were not related only to whey protein denaturation levels in the milks.     

Acid-induced gelation of milk protein concentrates with added pectin: Effect of casein micelle dissociation

The dynamics of the formation of the acid gel network for mixtures of milk protein concentrate (MPC) and low methoxyl amidated (LMA) pectin were studied using rheological measurements. The results as a function of pectin content and casein micelle integrity, from neutral pH to approximately pH 4.2, together with the microstructural changes observed in some of these systems, are presented.The gelation profiles of a mixture of 4% w/v MPC and LMA pectin (0–0.075% w/v) after the addition of 1.2% w/v glucono-δ-lactone showed a gradual decrease in the shear modulus with the incorporation of pectin. The effects of casein micelle integrity on casein–pectin interactions were studied, by preparing MPC dispersions containing various levels of micellar casein. A gradual change in the shear modulus, from a disrupting effect of pectin added to MPC, in which the casein micelles are intact, to a clear synergistic effect of pectin added to dissociated casein systems, was found in the acid-induced milk gels.     

Gravity separation of raw bovine milk: fat globule size distribution and fat content of milk fractions

This project determined effects of time and temperature on changes of fat globule size distribution and fat content in milk fractions during gravity separation. Fresh raw bovine milk was gravity separated at 4 or 15 degrees C. After 2, 6, 12, and 48 h, seven fractions, from bottom fraction (F1) to top fraction (F7), were successively drained from a separation column. Higher temperature resulted in a faster rate of fat separation. Within 2 h, large fat globules had already moved to the top, and the volume mean diameter of F7 increased from 3.13 microm (without separation) to 3.48 and 3.64 microm, respectively, at 4 and 15 degrees C. In F7, there was little change in globule size distribution after 2 h, but fat content continued to increase with separation time. The fat content of F7 reached 26.6% after 48 h at 4 degrees C, achieving a 58.8% creaming capacity. For F1 to F6, longer separation time resulted in smaller fat globule sizes and lower fat contents, especially for F1. After 48 h at 4 degrees C, the volume mean diameter of F1 decreased from 3.23 microm (without separation) to 1.16 microm, and fat content decreased from 3.75% (without separation) to 0.20%. Gravity separation may have unique applications in the dairy industry today. Its simplicity makes it an effective procedure for small-scale dairy product manufacturers to produce milks with a range of fat contents without using a centrifugal cream separator.     

乳脂肪球膜功能特性的研究进展

乳脂肪以脂肪球的形式存在,乳脂肪球膜是包裹在乳脂肪球周围的三层生物薄膜,具有很高的营养价值。随着食品科学研究的深入和分离技术的发展,乳脂肪球膜中的活性成分及其功能作用正在逐渐被揭示。乳脂肪球膜是含有蛋白质、磷脂、鞘脂、神经节苷脂、胆碱、唾液酸和胆固醇的混合物,这些成分是具有重要功能的食品成分,应用于配方食品生产。本文综述了乳脂肪球膜中常见的蛋白质、脂质及其生物活性,综述了近年来乳脂肪球膜及其成分在改善肠道健康、改善大脑发育、改善肥胖及相关并发症、改善老年人虚弱、抗癌、抗氧化和缓解疲劳等方面的体内研究和临床研究进展,并讨论了其可能的作用机制,以期为乳脂肪球膜配料的研发及其在配方食品中的应用提供借鉴和参考。     

Characterization of protein components of natural and heat-treated milk fat globule membranes

The proteins associated with the milk fat globule membrane (MFGM), isolated from early, mid and late season whole milks, were characterized using one- and two-dimensional SDS-PAGE under reducing and non-reducing conditions. In some experiments, MFGM separated from fresh whole milk was suspended in simulated milk ultrafiltrate (SMUF) and heated at various temperatures and times. SDS-PAGE under reducing conditions followed by staining with Coomassie blue showed the presence of about 37 protein bands, ranging in molecular weight from 15 to 200 kDa. SDS-PAGE under non-reducing conditions showed only about 25 distinct bands and the intensity of xanthine oxidase and butyrophilin bands was much less, while the intensity of PAS 6/7 band was similar compared with the reduced SDS-PAGE. Two-dimensional SDS-PAGE showed that the protein complexes that remained at the top of non-reducing gel were resolved into mostly xanthine oxidase and butyrophilin with a small proportion of PAS 6. These results indicate that xanthine oxidase and butyrophilin may be complexed via intermolecular disulfide bonds in the natural MFGM, although it is not possible to differentiate the individual protein distributions within these aggregates. It was found that the total protein content (mg/g fat) of MFGM and the percentage of xanthine oxidase and butyrophilin in early and late season MFGM were higher than that of mid season MFGM. In heated samples (above 60°C), xanthine oxidase and butyrophilin interacted further to form higher molecular weight protein complexes, while PAS 6/7 was relatively heat stable.     

Influence of heat treatment of goat milk on casein micelle size,rheological and textural properties of acid gels and set type yoghurts

Acid gels and yoghurts were made from goat milk that was heated at 72°C/30 s, 85°C/5 min, and 95°C/5 min, followed by acidification with starter culture at 43C until pH 4.6. The rheological and textural properties of acid gels and yoghurts were analyzed using dynamic low amplitude oscillatory rheology and back extrusion texture analysis, respectively. The effect of goat milk heat treatment on the mean casein micelle diameter and protein profile was also determined by dynamic light scattering and SDS PAGE electrophoresis, respectively. The shortest gelation and fermentation time was recorded for yoghurt prepared from milk heated at 85°C/5 min. Also, the pH of gelation, the storage moduli (G′) and yield stress were higher for this yoghurt, compared with the other two. Textural properties of goat milk yoghurts such as firmness and consistency were strongly affected by milk heat treatment, and the highest values were recorded for yoghurt produced from milk preheated at 85°C/5 min, as well. The largest casein micelles were measured after 85°C/5 min treatment and their size decreased at higher temperature, despite higher denaturation of whey proteins at the most intense heat regime, indicating the structure changes that influence on the acid gelation.     

Effects of milk protein polymorphism and composition,casein micelle size and salt distribution on the milk coagulation properties in Norwegian Red cattle

Effects of milk protein polymorphism and composition, casein micelle size and salts distribution on the coagulation properties of milk from 99 Norwegian Red cattle (NRF) were studied. Genetic variants of α_S1-casein (CN), β-CN, κ-CN and β-lactoglobulin (LG) affected rennet coagulation properties of milk. Significant effects of κ-CN and the composite genotype α_S1-β-κ-CN were observed on acid coagulation properties. Relative concentrations of milk proteins were significantly affected by individual casein genotypes and the composite genotype of α_S1-β-κ-CN while, the relative concentration of β-LG was only affected by β-LG genotypes. The salts distribution in milk and the concentration of milk proteins affected both rennet and acid coagulation properties. Milk protein genotypes associated with better rennet coagulation, impaired the acid coagulation properties. However, α_S1-β-κ-CN BB-A¹A²-BE and BB-A²A²-BB were associated with poor rennet and acid coagulation properties. Breeding programs should focus on decreasing these genotypes in NRF cattle.     

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

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

Variation in fat globule size in bovine milk and its prediction using mid-infrared spectroscopy

The objectives of this study were to investigate the sources of variation in milk fat globule (MFG) size in bovine milk and its prediction using mid-infrared (MIR) spectroscopy. Mean MFG size was measured in 2,076 milk samples from 399 Ayrshire, Brown Swiss, Holstein, and Jersey cows, and expressed as volume moment mean (D[4,3]) and surface moment mean (D[3,2]). The mid-infrared spectra of the samples and milk performance data were also recorded during routine milk recording and testing. The effects of breed, herd nested within breed, days in milk, season, milking period, age at calving, parity, and individual animal on the variation observed in MFG size were investigated. Breed, herd nested within breed, days in milk, season, and milking period significantly affected mean MFG size. Milk fat globule size was the largest at the beginning of lactation and subsequently decreased. Milk samples with the smallest MFG on average came from Holstein cows, and those with the largest were from Jersey and Brown Swiss cows. Partial least squares regression was used to predict MFG size from MIR spectra of samples with a calibration data set containing 2,034 and 2,032 samples for D[4,3] and D[3,2], respectively. Coefficients of determination of cross validation for D[4,3] and D[3,2] prediction models were 0.51 and 0.54, respectively. The associated ratio of performance deviation values were 1.43 and 1.48 for D[4,3] and D[3,2], respectively. With these models, individual mean MFG size could not be accurately predicted, but results may be sufficient to screen samples for having either small or large MFG on average. Significant but low correlations of D[4,3] and D[3,2] with milk fat yield were estimated (0.16 and 0.21, respectively). Significant and moderate Pearson correlation coefficients for fat percent with D[4,3] and D[3,2] were assessed (0.34 and 0.36, respectively). This correlation was greater between milk fat percentage and predicted MFG size than with measured MFG size with coefficients of 0.47 and 0.49 for D[4,3] and D[3,2], respectively. The MIR prediction equations are potentially overusing the correlation between fat and MFG size and exploiting the strong relationship between the MIR spectra and total milk fat. However, the predictions of MFG size are able to determine variation in mean globule size beyond what would be achieved just by looking at the correlation with fat production.     

The association of lipophilic phospholipids with native bovine casein micelles in skim milk: Effect of lactation stage and casein micelle size

A known biological role of casein micelles is to transport calcium from mother to young and provide amino acids for growth and development. Previous reports demonstrated that modified casein micelles can be used to transport and deliver hydrophobic probes. In this study, the distribution of lipid-soluble phospholipids, including sphingomyelins (SM) and phosphatidylcholines (PC), was quantified in whole raw milk, skim raw milk, and casein micelles of various sizes during early, mid, and late lactation stages. Low-pressure size exclusion chromatography was used to separate casein micelles by size, followed by hydrophobic extraction and liquid chromatography–mass spectrometry for the quantification of PC and SM. Results showed that the SM d18:1/23:0, d18:1/22:0, d18:1/16:0, d16:1/22:0, d16:1/23:0, and d18:1/24:0 and the PC 16:0/18:1, 18:0/18:2, and 16:0/16:0 were dominating candidates appearing in maximum concentration in whole raw milk obtained from late lactation, with 21 to 50% of total SM and 16 to 35% of total PC appearing in skim milk. Of the total SM and PC found in skim milk, 35 to 46% of SM and 22 to 29% of PC were associated with the casein micelle fraction. The highest concentrations of SM d18:1/22:0 (341 ± 17 µg/g of casein protein) and PC 16:0/18:1 (180 ± 20 µg/g of casein protein) were found to be associated with the largest casein micelles (diameter = 149 nm) isolated in milk from late lactation, followed by a decrease in concentration as the casein micelle size decreased.     

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.     

Temporal variation of milk fat globule diameter,fat and cholesterol content and milk epithelial cell gene expression in dairy cows

It was possible to show a connection between the temporal variation of milk fat globule diameter, fat and cholesterol content in milk and the expression of candidate genes in the mammary gland epithelial cells in milk. The beginning of lactation corresponded with higher levels of fat and cholesterol in the milk as a result of a higher expression of key enzymes in the purified bovine milk epithelial cells, paralleled with an increase in milk fat globule mean size.     

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.     

Heritabilities of measured and mid-infrared predicted milk fat globule size,milk fat and protein percentages,and their genetic correlations

The objective of this study was to estimate the heritability of milk fat globule (MFG) size and mid-infrared (MIR) predicted MFG size in Holstein cattle. The genetic correlations between measured and predicted MFG size with milk fat and protein percentage were also investigated. Average MFG size was measured in 1,583 milk samples taken from 254 Holstein cows from 29 herds across Canada. Size was expressed as volume moment mean (D[4,3]) and surface moment mean (D[3,2]). Analyzed milk samples also had average MFG size predicted from their MIR spectral records. Fat and protein percentages were obtained for all test-day milk samples in the cow's lactation. Univariate and bivariate repeatability animal models were used to estimate heritability and genetic correlations. Moderate heritabilities of 0.364 and 0.466 were found for D[4,3] and D[3,2], respectively, and a strong genetic correlation was found between the 2 traits (0.98). The heritabilities for the MIR-predicted MFG size were lower than those estimated for the measured MFG size at 0.300 for predicted D[4,3] and 0.239 for predicted D[3,2]. The genetic correlation between measured and predicted D[4,3] was 0.685; the correlation was slightly higher between measured and predicted D[3,2] at 0.764, likely due to the better prediction accuracy of D[3,2]. Milk fat percentage had moderate genetic correlations with both D[4,3] and D[3,2] (0.538 and 0.681, respectively). The genetic correlation between predicted MFG size and fat percentage was much stronger (greater than 0.97 for both predicted D[4,3] and D[3,2]). The stronger correlation suggests a limitation for the use of the predicted values of MFG size as indicator traits for true average MFG size in milk in selection programs. Larger samples sizes are required to provide better evidence of the estimated genetic parameters. A genetic component appears to exist for the average MFG size in bovine milk, and the variation could be exploited in selection programs.     

Milk quality and automatic milking: fat globule size, natural creaming, and lipolysis

Thirty-eight Italian Friesian first-lactation cows were allocated to 2 groups to evaluate the effect of 1) an automatic milking system (AMS) vs. milking in a milking parlor (MP) on milk fat characteristics; and 2) milking interval (≤480, 481 to 600, 601 to 720, and >720 min) on the same variables. Milk fat was analyzed for content (% vol/vol), natural creaming (% of fat), and free fatty acids (FFA, mEq/100 g of fat). Distribution of milk fat globule size was evaluated to calculate average fat globule diameter (d₁), volume-surface average diameter (d₃₂), specific globule surface area, and mean interglobular distance. Milk yield was recorded to calculate hourly milk and milk fat yield. Milking system had no effect on milk yield, milk fat content, and hourly milk fat yield. Milk from AMS had less natural creaming and more FFA content than milk from MP. Fat globule size, globular surface area, and interglobular distance were not affected by milking system per se. Afternoon MP milkings had more fat content and hourly milk fat yield than AMS milkings when milking interval was >480 min. Milk fat FFA content was greater in AMS milkings when milking interval was ≤480 min than in milkings from MP and from AMS when milking interval was >600 min. Milking interval did not affect fat globule size, expressed as d₃₂. Results from this experiment indicate a limited effect of AMS per se on milk fat quality; a more important factor seems to be the increase in milking frequency, generally associated with AMS.