A method for the large-scale isolation of β-casein

A method for the large-scale isolation of β-casein from renneted skim milk was developed. The curd from renneted skim milk was dispersed in hot (?70 °C) water to inactivate residual chymosin. The heated curd was subsequently recovered by centrifugation, resuspended in water and incubated at 5 °C, during which β-casein dissociated from the curd; the suspension was centrifuged and the aqueous phase lyophilised. The isolated protein consisted mainly of β-casein, containing a minor amount of γ-caseins and traces of other caseins. Unless chymosin was fully inactivated by heating, some β-casein was hydrolysed at the Leu₁₉₂–Tyr₁₉₃ bond. The yield of β-casein increased with incubation time, up to ∼20% of the β-casein present in the milk after 24 h at 5 °C. Reducing milk pH to 5.5 or 6.0, prior to renneting, caused a high level of contamination with α_s-caseins. This isolation procedure can be easily scaled-up to an industrial process and the β-casein-depleted curd may be used for the manufacture of rennet casein or processed cheese.     

Milk protein fractions strongly affect the patterns of coagulation,curd firming,and syneresis

The aim of this study was to assess the role of milk protein fractions in the coagulation, curd firming, and syneresis of bovine milk. Analyses were performed on 1,271 individual milk samples from Brown Swiss cows reared in 85 herds classified into 4 types of farming systems, from the very traditional (tied cows, feed manually distributed, summer highland pasture) to the most modern (loose cows, use of total mixed rations with or without silage). Fractions α_S1-casein (CN), α_S2-CN, β-CN, κ-CN, β-lactoglobulin (LG), and α-lactalbumin (LA) and genotypes at CSN2, CSN3, and BLG were obtained by reversed-phase HPLC. The following milk coagulation properties were measured with a lactodynamograph, with the testing time extended to 60 min: rennet coagulation time (RCT, min), curd firming time (min), and curd firmness at 30 and 45 min (mm). All the curd firmness measures recorded over time (total of 240 observations/sample) were used in a 4-parameter nonlinear model to obtain parameters of coagulation, curd firming, and syneresis: RCT estimated from the equation (min), asymptotic potential curd firmness (mm), the curd firming and syneresis instant rate constants (%/min), and the maximum curd firmness value (CF_max, mm) and the time taken to reach it (min). All the aforementioned traits were analyzed with 2 linear mixed models, which tested the effects of the protein fractions expressed in different ways: in the first, quantitative model, each protein fraction was expressed as content in milk; in the second, qualitative model, each protein fraction was expressed as a percentage of total casein content. Besides proteins, additional nuisance parameters were herd (included as a random effect), daily milk production (only for the quantitative model), casein content (only for the qualitative model), dairy system, parity, days in milk, the pendulum of the lactodynamograph, and the CSN2, CSN3, and BLG genotypes. Both α_S1-CN and β-CN showed a clear and favorable effect on CF_max, where the former effect was almost double the latter. Milk coagulation ability was favorably affected by κ-CN, which reduced both the RCT and RCT estimated from the equation, increased the curd firming and syneresis instant rate constants, and allowed a higher CF_max to be reached. In contrast, α_S2-CN delayed gelation time and β-LG worsened curd firming, both resulting in a low CF_max. The results of this study suggest that modification of the relative contents of specific protein fractions can have an enormous effect on the technological behavior of bovine milk.     

Enzymatic modification of protein preparation obtained from water-washed mechanically recovered poultry meat

Studies were conducted on a protein preparation obtained from washed mechanically recovered poultry meat (MRPM). The effect of addition of 3 g/kg microbial transglutaminase (MTG) to poultry meat protein was evaluated in terms of texture changes by dynamic mechanical analysis (DMA) and nuclear magnetic resonance (NMR) to determine water content in the preparation and its effect on protein. Samples with the addition of MTG were pre-incubated at 5–6 °C for 1.5, 3, 4.5, 6, 7.5, 9 and 24 h. The largest changes for both texture parameters and rheological properties were observed in the interval of approx. 4–7 h incubation. The protein preparation with the enzyme added had significantly higher values of the moduli of elasticity (G₁) and losses (G₂) in comparison to the control system. Samples with the addition of MTG also showed a higher water-binding capacity. From the NMR studies it was found that the greatest amount of water was bound by protein in the period of approx. 2.5–5 h incubation. After that time an increase was found in the amount of free water in the sample, which suggests that it was displaced from the system by stronger protein–protein bonds.     

Variations in milk protein fractions affect the efficiency of the cheese-making process

The aim of this study was to assess the influence of the amounts of the α_S1-, α_S2-, β-, and κ-casein (CN) and the α-lactalbumin and β-lactoglobulin protein fractions on the efficiency of the cheese-making process independently of their genetic polymorphisms. The study was carried out on milk samples from 1,271 Brown Swiss cows from 85 herds classified into 4 categories according to management, feeding, and housing characteristics (traditional and modern systems). To assess the efficiency of the cheese-making process, we processed the milk samples according to a laboratory cheese-making procedure (1,500 mL/sample) and obtained the following measures: (1) 3 percentage cheese yields (%CY_curd, %CY_solids, %CY_water), (2) 2 daily cheese yields obtained by multiplying %CY (curd and total solids) by daily milk yields (dCY_curd, dCY_solids), (3) 4 measures of nutrient recovery in the curd (REC_fat, REC_protein, REC_solids, REC_energy), and (4) 2 measures of cheese-making efficiency in terms of the ratio between the observed and theoretical %CY (Ef-%CY_curd, Ef-%CY_solids). All the aforementioned traits were analyzed by fitting 2 linear mixed models with protein fractions as fixed effects expressed as percentage in the milk (model M-%milk) and as percentage of the total casein content (model M-%cas) together with the effects of total casein content (only in model M-%cas), daily milk yield (only in model M-%milk; not for dCY traits), dairy system, herd (random effect), days in milk, parity, and vat. The efficiency of overall cheese yield (Ef-%CY_curd) was mostly positively associated with β-CN content in the milk, whereas Ef-%CY_solids was greater with higher amounts of κ-CN and α_S1-CN (M-%milk) due to the strong influence of both fractions on the recovery rate of milk components in the curd (fat and total solids, protein with α_S1-CN only) when expressed as percentage of milk and of total casein; only β-CN was more important for REC_protein. In contrast, we found β-lactoglobulin to be highly negatively related to all the traits related to the cheese-making process and to the daily cheese yield per cow, whereas α-lactalbumin was positively associated with the latter traits. Additional research on this topic is needed, with particular focus on the genetic and genomic aspects of the role of protein fractions in the cheese-making process and on the associations between genetic polymorphisms in milk protein and milk nutrient recovery in the curd.     

Effect of sesame protein isolate in partial replacement of milk protein on the rheological,textural and microstructural characteristics of fresh cheese

Soft cheeses were manufactured from bovine milk with the addition of 0–12% sesame protein isolate (SPI) were utilised to investigate rheology, texture and microstructure at different stages of cheese making. SPI addition reduced the speed of milk fermentation, kappa‐casein proteolysis of rennet and elongated the time of cheese curd formation. Renneted milk storage modulus G′_60min was decreased and coagulation time increased with increasing SPI content. Low SPI supplements (4% and 8%) enhanced the hardness, cohesiveness, adhesiveness and gumminess of the soft cheese, while high SPI addition (12%) deteriorated the texture. In the cheese curd gel matrix, SPI distributed as specific SPI‐gel clusters on the surface of curd fractures, stacked or fused with ball‐shaped casein micelles and wrapped up to casein gel strands. In summary, SPI actively interacted with casein colloid throughout the cheese making process.     

Associations between pathogen-specific cases of subclinical mastitis and milk yield,quality, protein composition,and cheese-making traits in dairy cows

The aim of this study was to investigate associations between pathogen-specific cases of subclinical mastitis and milk yield, quality, protein composition, and cheese-making traits. Forty-one multibreed herds were selected for the study, and composite milk samples were collected from 1,508 cows belonging to 3 specialized dairy breeds (Holstein Friesian, Brown Swiss, and Jersey) and 3 dual-purpose breeds of Alpine origin (Simmental, Rendena, and Grey Alpine). Milk composition [i.e., fat, protein, casein, lactose, pH, urea, and somatic cell count (SCC)] was analyzed, and separation of protein fractions was performed by reversed-phase high performance liquid chromatography. Eleven coagulation traits were measured: 5 traditional milk coagulation properties [time from rennet addition to milk gelation (RCT, min), curd-firming rate as the time to a curd firmness (CF) of 20 mm (k₂₀, min), and CF at 30, 45, and 60 min from rennet addition (a₃₀, a₄₅, and a₆₀, mm)], and 6 new curd firming and syneresis traits [potential asymptotical CF at an infinite time (CF_P, mm), curd-firming instant rate constant (k_CF, % × min^?1), curd syneresis instant rate constant (k_SR, % × min^?1), modeled RCT (RCT_eq, min), maximum CF value (CF_max, mm), and time at CF_max (t_max, min)]. We also measured 3 cheese yield traits, expressing the weights of total fresh curd (%CY_CURD), dry matter (%CY_SOLIDS), and water (%CY_WATER) in the curd as percentages of the weight of the processed milk, and 4 nutrient recovery traits (REC_PROTEIN, REC_FAT, REC_SOLIDS, and REC_ENERGY), representing the percentage ratio between each nutrient in the curd and milk. Milk samples with SCC > 100,000 cells/mL were subjected to bacteriological examination. All samples were divided into 7 clusters of udder health (UH) status: healthy (cows with milk SCC < 100,000 cells/mL and uncultured); culture-negative samples with low, medium, or high SCC; and culture-positive samples divided into contagious, environmental, and opportunistic intramammary infection (IMI). Data were analyzed using a linear mixed model. Significant variations in the casein to protein ratio and lactose content were observed in all culture-positive samples and in culture-negative samples with medium to high SCC compared to normal milk. No differences were observed among contagious, environmental, and opportunistic pathogens, suggesting an effect of inflammation rather than infection. The greatest impairment in milk quantity and composition, clotting ability, and cheese production was observed in the 2 UH status groups with the highest milk SCC (i.e., contagious IMI and culture-negative samples with high SCC), revealing a discrepancy between the bacteriological results and inflammatory status, and thus confirming the importance of SCC as an indicator of udder health and milk quality.     

Genetic parameters of different measures of cheese yield and milk nutrient recovery from an individual model cheese-manufacturing process

Cheese yield (CY) is an important technological trait in the dairy industry, and the objective of this study was to estimate the genetic parameters of cheese yield in a dairy cattle population using an individual model-cheese production procedure. A total of 1,167 Brown Swiss cows belonging to 85 herds were sampled once (a maximum of 15 cows were sampled per herd on a single test day, 1 or 2 herds per week). From each cow, 1,500 mL of milk was processed according to the following steps: milk sampling and heating, culture addition, rennet addition, gelation-time recording, curd cutting, whey draining and sampling, wheel formation, pressing, salting in brine, weighing, and cheese sampling. The compositions of individual milk, whey, and curd samples were determined. Three measures of percentage cheese yield (%CY) were calculated: %CY_CURD, %CY_SOLIDS, and %CY_WATER, which represented the ratios between the weight of fresh curd, the total solids of the curd, and the water content of the curd, respectively, and the weight of the milk processed. In addition, 3 measures of daily cheese yield (dCY, kg/d) were defined, considering the daily milk yield. Three measures of nutrient recovery (REC) were computed: REC_FAT, REC_PROTEIN, and REC_SOLIDS, which represented the ratio between the weights of the fat, protein, and total solids in the curd, respectively, and the corresponding nutrient in the milk. Energy recovery, REC_ENERGY, represented the energy content of the cheese versus that in the milk. For statistical analysis, a Bayesian animal model was implemented via Gibbs sampling. The effects of parity (1 to ≥4), days in milk (6 classes), and laboratory vat (15 vats) were assigned flat priors; those of herd-test-date, animal, and residual were given Gaussian prior distributions. Intra-herd heritability estimates of %CY_CURD, _%CY_SOLIDS, and %CY_WATER ranged from 0.224 to 0.267; these were larger than the estimates obtained for milk yield (0.182) and milk fat content (0.122), and similar to that for protein content (0.275). Daily cheese yields showed heritability estimates similar to those of daily milk yield. The trait %CY_WATER showed a highly positive genetic correlation with %CY_SOLIDS (0.87), whereas their phenotypic correlation was moderate (0.37), and the fat and protein contents of milk showed high genetic correlations with %CY traits. The heritability estimates of REC_PROTEIN and REC_FAT were larger (0.490 and 0.208, respectively) than those obtained for the protein and fat contents of milk, and the genetic relationships between REC_PROTEIN and REC_FAT with milk protein and fat content were low or moderate; REC_PROTEIN and REC_FAT were moderately correlated with the %CY traits and highly correlated with REC_SOLIDS and REC_ENERGY. Both REC_SOLIDS and REC_ENERGY were heritable (0.274 and 0.232), and showed high correlations with each other (0.96) and with the %CY traits (0.83 to 0.97). Together, these findings demonstrate the existence of economically important, genetically determined variability in cheese yield that does not depend solely upon the fat and protein contents of milk, but also relies on the ability of the coagulum to retain the highest possible proportions of the available protein, fat, and water. Exploitation of this interesting genetic variation does not seem to be feasible through direct measurement of the phenotype in cows at the population level. Instead, further research is warranted to examine possible means for indirect prediction, such as through assessing the mid-infrared spectra of milk samples.     

Improving the yield of Mozzarella cheese by phospholipase treatment of milk

Part-skim Mozzarella cheese was manufactured from milk hydrolyzed with fungal phospholipase A₁ prior to renneting. The phospholipase treatment reduced fat losses in whey and cooking water and increased cheese yield as a result of improved fat and moisture retention in the cheese curd. The amount of phospholipids in the whey was reduced because of improved retention of lysophospholipids in the cheese curd. Water binding in the fresh curds and young cheeses up to 3 wk of storage was investigated by a ¹H nuclear magnetic resonance spin-spin relaxation technique. In the fresh curds, 2 dominant water fractions were present, characterized by average spin-spin relaxation times (T₂) of 14 and 86 to 89 ms, respectively. These 2 fractions of low- and high-molecular-mobility water were similar in all cheeses and presumed to represent water associated with the casein matrix and water present in the pores. A few hours after manufacture, cheeses made with phospholipase showed decreased T₂ of the high-mobility fraction, indicating improved water-holding capacity. It is suggested that lysophospholipids released from the fat globule membranes act as surface-active agents in the cheese curd, helping emulsification of water and fat during processing and reducing syneresis. During 3 wk of storage after manufacture, the mobility of both water fractions increased in all cheeses, but was highest in the cheeses made with phospholipase. The increase in mobility during the first weeks of storage has earlier been ascribed to structural changes in the protein matrix, which in principle could be accelerated because of the higher moisture content. However, the microstructure of phospholipase-treated cheese was investigated by confocal laser scanning microscopy and found to be very similar to the control cheese during processing and up to 28 d of storage. In addition, flowability, stretchability, and browning were acceptable and similar in all the manufactured cheeses. Thus, phospholipase hydrolysis of cheese milk improved the cheese yield without changing the cheese microstructure, and resulted in cheese with functional properties that were identical to traditional Mozzarella cheese.     

Characterization of Cheese Curd Ripened with Penicillium caseicolum for Producing a Flavor Concentrate

Camembert cheese was prepared from homogenized milk containing veal oral lipase and ripened in a loose curd form to speed up the ripening process. Traditional hooped cheese was a control. Free fatty acid titer increased from 58 μmole in the fresh cheese to 359 μmole in control cheese and 2,289 μmole/g of cheese fat in loose curd cheese. After 3 wk of ripening, the cheese had 70% of its total protein as water soluble protein as compared to 39% in the hooped cheese. Polyacrylamide gel electrophoresis showed 86% of α_S1-caseins and 35% of β-casein had been degraded in the loose curd cheese. Sensory analysis demonstrated that the loose curd process rapidly produced cheese with intense flavor.     

CHANGES IN Pb, Cd, Fe, Cu AND Zn LEVELS DURING THE PRODUCTION OF KAŞAR CHEESE

ABSTRACT

Levels of Pb, Cd, Fe, Cu and Zn in milk, curd, pressed curd, fresh cheese, whey, rennet and scalding water taken from two different Ka?ar cheese plants (A and B) in Ankara, Turkey were investigated. The milk used in plant A contained higher amount of Pb, Fe and Zn than the milk used in plant B. Pb level during processing in both dairy plants showed a significant increase from milk to curd (626.2–912.3 µg/kg for plant A and 265.2–371.8 µg/kg, dry weight, for plant B) (P < 0.01). Similarly, Fe, Cu and Zn contents of the curds in plant A and B showed an important increase with respect to the milk (P < 0.01). During transition of the curd to pressed curd and of pressed curd to fresh cheese, almost all metals tested showed a decrease because of the loss of these metals into whey and scalding water. The results showed that curdling the milk was the most important contamination step.

PRACTICAL APPLICATIONS

Heavy metals may enter the human body through food, water, air or absorption through the skin, and can cause metabolic anormalies. Heavy metals may reach our foods from a number of sources. The more important of these are: soil; the chemicals applied to agricultural land; the water used in food processing or cooking; and the equipment, containers and utensils used for food processing, storage or cooking. Milk and milk products are the basic components of the human diet, and among milk products cheese holds an important place. Ka?ar cheese ranks second with respect to consumption, significantly contributing to the Turkish diet. In order to prevent the health risk of consuming contaminated cheeses, it is very important to determine the effect of equipment and process variation on the heavy‐metal content of Ka?ar cheese. The results of this study would help the regulatory authorities to establish a Hazard Analysis and Critical Control Points (HACCP) plan and to identify important contamination sources for Ka?ar and similar kind of cheeses.

     

Fate of aflatoxin M1 in cheese whey processing

Aflatoxin M₁ (AFM₁) is an important mycotoxin frequently found in milk and in dairy products. It is a minor metabolic product of Aspergillus flavus and A parasiticus. However, it occurs in dairy products as a metabolite formed in cows from aflatoxin B₁ contained in animal feeds. In cheese production, AFM₁ distributes between curd and whey, being present in products derived from cheese whey processing. In this study, cheese whey from dairy processing was artificially contaminated with the mycotoxin at about 0.1 µg l⁻¹. Ultra‐filtration experiments of whey were carried out in order to determinate AFM₁ distribution between retentate (protein‐rich fraction) and permeate (lactose‐rich fraction). Recoveries of AFM₁ in retentate were 72.6–86.4% while, in permeate, recoveries were in the range 2.4–14.7%. Partition coefficients of AFM₁, lactose and protein were calculated to determine whether there was an interaction between AFM₁ and protein. In all experiments, AFM₁ partition coefficient was lower than 1, whilst for lactose coefficients close to 1 were determined, showing an affinity of aflatoxin M₁ to the protein‐rich fraction (retentate). Copyright © 2005 Society of Chemical Industry     

Variation of milk technological properties in sheep milk: Relationships among composition,coagulation and cheese-making traits

The relationships between milk composition, coagulation properties and cheese-making traits in sheep milk were characterised. Ten traits related to milk coagulation (RCT_eq, k_CF, CF_p), cheese yield (%CY_CURD, %CY_SOLIDS, %CY_WATER), and curd nutrients recovery or whey loss (%REC_FAT, %REC_PROTEIN, %REC_SOLIDS, %REC_ENERGY) were recorded. To obtain a measure of the efficiency in terms of %CY, the ratio between the observed and the theoretical %CY (Ef-%CY_CURD, Ef-%CY_SOLIDS) was calculated. Sheep milk showed good qualities for coagulation and cheese production; milk lactose appeared to be the component most linked to gelation, curd firming time and water retained in the curd. In the case of milk protein, an opposite relationship with gelation time was observed. Milk fat and protein positively affected total solids recovery and yield inducing higher %CY_CURD. Relationships with CF_t parameters were limited; curd firming instant rate seems to be the most informative trait to assess the efficiency of the cheese-making process.     

Preliminary screening of Bifidobacteria spp. and Pediococcus acidilactici in a Swiss cheese curd slurry model system: Impact on microbial viability and flavor characteristics

A Swiss cheese curd slurry model system was used as a preliminary screening method to determine the feasibility of the incorporation of probiotic bacteria (Bifidobacterium breve R0070, Bifidobacterium infantis R0033, Bifidobacterium longum R0175, and Pediococcus acidilactici R1001) into Swiss cheese. The cheese curd was inoculated with probiotic bacteria (8.0 log₁₀ cfu g⁻¹) and ripened anaerobically for 0, 7, and 10 days at 37 °C. Following ripening, counts of the probiotic bacteria increased to 9–10 log₁₀ cfu g⁻¹, with no significant difference in the viability of the four probiotic bacteria. Viable populations of Swiss cheese background microflora in the presence of each probiotic culture were comparable with the control. Ripening time, and to a lesser extent probiotic treatment had a significant effect on the content of several volatile flavor compounds. Similarly, ripening time contributed to a significant increase in the content of a majority of the free amino acids. The study demonstrated the feasibility of the incorporation of probiotic bacteria into Swiss cheese to produce a functional food.     

Goat farm variability affects milk Fourier-transform infrared spectra used for predicting coagulation properties

Driven by the large amount of goat milk destined for cheese production, and to pioneer the goat cheese industry, the objective of this study was to assess the effect of farm in predicting goat milk-coagulation and curd-firmness traits via Fourier-transform infrared spectroscopy. Spectra from 452 Sarda goats belonging to 14 farms in central and southeast Sardinia (Italy) were collected. A Bayesian linear regression model was used, estimating all spectral wavelengths' effects simultaneously. Three traditional milk-coagulation properties [rennet coagulation time (min), time to curd firmness of 20 mm (min), and curd firmness 30 min after rennet addition (mm)] and 3 curd-firmness measures modeled over time [rennet coagulation time estimated according to curd firmness change over time (RCT_eq), instant curd-firming rate constant, and asymptotical curd firmness] were considered. A stratified cross validation (SCV) was assigned, evaluating each farm separately (validation set; VAL) and keeping the remaining farms to train (calibration set) the statistical model. Moreover, a SCV, where 20% of the goats randomly taken (10 replicates per farm) from the VAL farm entered the calibration set, was also considered (SCV₈₀). To assess model performance, coefficient of determination (R²_VAL) and the root mean squared error of validation were recorded. The R²_VAL varied between 0.14 and 0.45 (instant curd-firming rate constant and RCT_eq, respectively), albeit the standard deviation was approximating half of the mean for all the traits. Although average results of the 2 SCV procedures were similar, in SCV₈₀, the maximum R²_VAL increased at about 15% across traits, with the highest observed for time to curd firmness of 20 mm (20%) and the lowest for RCT_eq (6%). Further investigation evidenced important variability among farms, with R²_VAL for some of them being close to 0. Our work outlined the importance of considering the effect of farm when developing Fourier-transform infrared spectroscopy prediction equations for coagulation and curd-firmness traits in goats.     

Occurrence of β-casein fragments in cold-stored and curdled river buffalo (Bubalus bubalis L.) milk

The safeguard of river buffalo Mozzarella cheese, a Protected Designation of Origin dairy product, has prompted an analytical study to trace the milk and curd used as raw material in cheesemaking. This is to prevent the illegal use of milk or curd from different geographical areas outside of those indicated in the official production protocol. For this purpose, we studied primary proteolysis occurring in fresh and frozen milk and curd to identify a molecular marker that could indicate the raw material used. Whole casein from frozen river buffalo milk was separated using cation-exchange chromatography and sodium dodecyl sulfate-PAGE, and a protein component with an estimated molecular weight of 15.3 kDa was detected. This protein component was revealed in fresh river buffalo milk as a faint electrophoresis band, which drastically increased in intensity in refrigerated and frozen milk as well as in curd and was found to be associated with β-CN through hydrophobic interaction. By using matrix-assisted laser desorption/ionization-time of flight peptide mass mapping, this component was identified as the C-terminal fragment f(69-209) of β-CN (expected molecular weight of 15,748.8 Da). β-Casein f(69-209), originating from the early hydrolysis of Lys₆₈-Ser₆₉ by plasmin, has no counterpart in bovine milk. The increased rate of hydrolysis by plasmin toward the cleavage site Lys₆₈-Ser₆₉ has to be ascribed to the elevated proline content of the peptide 61-73. The favored production of β-CN f(69-209) has also drawn attention to the complementary proteose peptone β-CN f(1-68) that is presumed to play a physiological role in inducing milk secretion similar to that of β-CN f(1-29). The higher in vivo and in vitro production rate, compared with γ₁-CN formation, indicates that β-CN f(69-209) and its complementary fragment are candidate molecular markers to evaluate milk and curd freshness. We used indirect ELISA analysis based on the determination of remaining nonhydrolyzed β-CN to perform a quantitative evaluation of proteolysis.     

Saccharomyces cerevisiae improves rennet-induced gelation of ultra-high temperature milk

Heating milk at high temperatures impairs its renneting properties, but rennet-induced curds can be formed from ultra-high temperature (UHT) milk inoculated with Saccharomyces cerevisiae. Herein, we measured physicochemical indices of UHT milk inoculated with S. cerevisiae before rennet addition, monitored the kinetics of gel formation, and investigated the physicochemical properties and microstructure of rennet-induced curds to explore the mechanisms by which S. cerevisiae influenced rennet-induced gelation of UHT milk. Compared with untreated pasteurized cow milk and UHT milk, the ethanol content was increased, the pH was decreased, the particle size and ζ-potential were increased, the time points at which the elasticity index began to increase were advanced, and the maximum elasticity index was increased for UHT milk inoculated with S. cerevisiae. The number of S. cerevisiae cells affected the structure of rennet-induced curds; with few cells added, the protein network of curds was continuous and tight, the mean square displacement curves showed an asymptotic behavior, and the water retention capacity and curd yield were high; with more cells added, the loosely entangled proteins aggregated, the continuity of the network was destroyed, and the curd yield decreased. In summary, a low number of S. cerevisiae cells (<1.0 × 10⁷ cfu/mL) can increase particle size, ζ-potential, and ethanol content, and decrease pH of S. cerevisiae-inoculated UHT milk, thereby accelerating the aggregation reactions after enzymatic reaction and improving the renneting properties.     

Effects of the detailed protein composition of milk on curd yield and composition measured by model micro-cheese curd making of individual milk samples

The effect of the contents of casein (CN) and whey protein fractions on curd yield (CY) and composition was estimated using 964 individual milk samples. Contents of α_S1-CN, α_S2-CN, β-CN, γ-CN, glycosylated κ-CN (Gκ-CN), unglycosylated κ-CN, β-LG, and α-LA of individual milk samples were measured using reversed-phase HPLC. Curd yield and curd composition were measured by model micro-cheese curd making using 25 mL of milk. Dry matter CY (DMCY) was positively associated with all casein fractions but especially with α_S1-CN and β-CN. Curd moisture decreased at increasing β-CN content and increased at increasing γ-CN and Gκ-CN content. Due to their associations with moisture, Gκ-CN and β-CN were the fractions with the greatest effect on raw CY, which decreased by 0.66% per 1-standard deviation (SD) increase in the content of β-CN and increased by 0.62% per 1-SD increase in the content of Gκ-CN. The effects due to variation in percentages of the casein fractions in total casein were less marked than those exerted by contents. A 1-SD increase in β-CN percentage in casein (+3.8% in casein) exerted a slightly negative effect on DMCY (β = ?0.05%). Conversely, increasing amounts of α_S1-CN percentage were associated with a small increase in DMCY. Hence, results suggest that, at constant casein and whey protein contents in milk, the DMCY depends to a limited extent on the variation in the α_S1-CN:β-CN ratio. κ-Casein percentage did not affect DMCY, indicating that the positive relationship detected between the content of κ-CN and DMCY can be attributed to the increase in total casein resulting from the increased amount of κ-CN and not to variation in κ-CN relative content. However, milk with increased Gκ-CN percentage in κ-CN also shows increased raw CY and produces curds with increased moisture content. Curd yield increased at increasing content and relative proportion of β-LG in whey protein, but this is attributable to an improved capacity of the curd to retain water. Results obtained in this study support the hypothesis that, besides variation in total casein and whey protein contents, variation in protein composition might affect the cheese-making ability of milk, but this requires further studies.     

Pathway-based genome-wide association analysis of milk coagulation properties,curd firmness,cheese yield,and curd nutrient recovery in dairy cattle

It is becoming common to complement genome-wide association studies (GWAS) with gene-set enrichment analysis to deepen the understanding of the biological pathways affecting quantitative traits. Our objective was to conduct a gene ontology and pathway-based analysis to identify possible biological mechanisms involved in the regulation of bovine milk technological traits: coagulation properties, curd firmness modeling, individual cheese yield (CY), and milk nutrient recovery into the curd (REC) or whey loss traits. Results from 2 previous GWAS studies using 1,011 cows genotyped for 50k single nucleotide polymorphisms were used. Overall, the phenotypes analyzed consisted of 3 traditional milk coagulation property measures [RCT: rennet coagulation time defined as the time (min) from addition of enzyme to the beginning of coagulation; k₂₀: the interval (min) from RCT to the time at which a curd firmness of 20 mm is attained; a₃₀: a measure of the extent of curd firmness (mm) 30 min after coagulant addition], 6 curd firmness modeling traits [RCT_eq: RCT estimated through the CF equation (min); CF_P: potential asymptotic curd firmness (mm); k_CF: curd-firming rate constant (% × min^?1); k_SR: syneresis rate constant (% × min^?1); CF_max: maximum curd firmness (mm); and t_max: time to CF_max (min)], 3 individual CY-related traits expressing the weight of fresh curd (%CY_CURD), curd solids (%CY_SOLIDS), and curd moisture (%CY_WATER) as a percentage of weight of milk processed and 4 milk nutrient and energy recoveries in the curd (REC_FAT, REC_PROTEIN, REC_SOLIDS, and REC_ENERGY calculated as the % ratio between the nutrient in curd and the corresponding nutrient in processed milk), milk pH, and protein percentage. Each trait was analyzed separately. In total, 13,269 annotated genes were used in the analysis. The Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway databases were queried for enrichment analyses. Overall, 21 Gene Ontology and 17 Kyoto Encyclopedia of Genes and Genomes categories were significantly associated (false discovery rate at 5%) with 7 traits (RCT, RCT_eq, k_CF, %CY_SOLIDS, REC_FAT, REC_SOLIDS, and REC_ENERGY), with some being in common between traits. The significantly enriched categories included calcium signaling pathway, salivary secretion, metabolic pathways, carbohydrate digestion and absorption, the tight junction and the phosphatidylinositol pathways, as well as pathways related to the bovine mammary gland health status, and contained a total of 150 genes spanning all chromosomes but 9, 20, and 27. This study provided new insights into the regulation of bovine milk coagulation and cheese ability that were not captured by the GWAS.     

Analysis of texture and dynamics of water binding in enzymatically modified myofibrillar preparation obtained from washed mechanically recovered poultry meat

Experiments were conducted on a myofibrillar preparation, obtained from washed mechanically recovered poultry meat. An enzymatic preparation containing microbial transglutaminase (MTG) was added to samples of the myofibril isolate. The binding of water contained in the protein preparation with added MTG was assessed using low field nuclear magnetic resonance (NMR). Moreover, texture was analyzed in myofibril samples with the addition of transglutaminase pre-incubated for 0.5, 1.5, 3, 4.5 and 6 h. All measurements were taken at 7 ± 0.2 °C. Samples with added transglutaminase exhibited improved mechanical failure strength and better water binding capacity. The most dynamic increase of texture parameter values was observed in the interval from 1.5 to 3 h pre-incubation of the preparation with the added enzyme. Based on NMR (T ₁) testing it was established that the highest amount of water was bound by protein in the period from approximately 1 to 1.5 h pre-incubation. After that time free water content in the sample was again found to increase. This means that water was displaced from the system as a result of protein–protein interactions dominating over protein–water interactions. The above suggests that the enzymatic modification of the protein preparation contributed to the intensification of cross-linking between proteins in the preparation.     

Phenotypic factors affecting coagulation properties of milk from Sarda ewes

In this study, milk-coagulation properties (MCP) were characterized in the Sarda sheep breed. Milk composition and MCP [rennet-coagulation time (RCT), curd-firming time [time to reach a curd firmness of 20 mm (k₂₀)], and curd firmness (a₃₀), (a₄₅), and (a₆₀)] were obtained extending the lactodynamographic analysis from 30 to 60 min from a population of 1,121 ewes from 23 different farms. Managerial characteristics of farms and parity, individual daily milk yields and stage of lactation of ewes were recorded. Data were analyzed using a mixed-model procedure with fixed effects of days in milk, parity, daily milk yield, and flock size and the random effect of the flock/test day nested within flock size. Sampled farms were classified as small (<300 ewes) and medium (300 to 600 ewes), and these were kept by family operations, or as large (>600 ewes), often operated through hired workers. Daily milk yield was, on average, 1.58 ± 0.79 L/d and variability for this trait was very high. The average content of fat, protein, and casein was respectively 6.41, 5.39, and 4.20%. The class of flock size had a significant effect only on curd firmness, whereas days in milk affected RCT and k₂₀. The flock test day, parity, and daily milk yield were important sources of variation for all MCP. The mean value of RCT (8.6 min) and the low occurrence of noncoagulating samples (0.44%) confirmed the excellent coagulation ability of sheep milk compared with cattle milk. A more rapid coagulation was observed in mid-lactating, primiparous, and high-yielding ewes. The k₂₀ was usually reached in less than 2 min after gelation, with the most favorable values at mid lactation. The mean value of curd firmness 30 min after rennet addition (a₃₀) was, on average, 50 mm and decreased to 46 and 42 mm respectively after 45 (a₄₅) and 60 min (a₆₀). The decreasing value of curd-firmness traits was likely to be caused by curd syneresis and whey expulsion. The correlation between RCT and a₃₀ was much lower than in dairy cows and about null for a₄₅ and a₆₀. This means that curd firmness in dairy ewes is almost independent of gelation time and this can provide specific information for this species. In conclusion, this study showed that milk from Sarda sheep is characterized by an earlier gelation, a faster increase in curd firmness with time, and greater curd firmness after 30 min compared with dairy cows. Furthermore, correlations between MCP in sheep are much lower than in bovines and some of the assumptions and interpretations related to cows cannot be applied to sheep.