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

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

Insoluble calcium content and rheological properties of Colby cheese during ripening

Colby cheese was made using different manufacturing conditions (i.e., varying the lactose content of milk and pH values at critical steps in the cheesemaking process) to alter the extent of acid development and the insoluble and total Ca contents of cheese. Milk was concentrated by reverse osmosis (RO) to increase the lactose content. Extent of acid development was modified by using high (HPM) and low (LPM) pH values at coagulant addition, whey drainage, and curd milling. Total Ca content was determined by atomic absorption spectroscopy, and the insoluble (INSOL) Ca content of cheese was measured by the cheese juice method. The rheological and melting properties of cheese were measured by small amplitude oscillatory rheometry and UW-Melt Profiler, respectively. There was very little change in pH during ripening even in cheese made from milk with high lactose content. The initial (d 1) cheese pH was in the range of 4.9 to 5.1. The INSOL Ca content of cheese decreased during the first 4 wk of ripening. Cheeses made with the LPM had lower INSOL Ca content during ripening compared with cheese made with HPM. There was an increase in melt and maximum loss tangent values during ripening except for LPM cheeses made with RO-concentrated milk, as this cheese had pH <4.9 and exhibited limited melt. Curd washing reduced the levels of lactic acid produced during ripening and resulted in significantly higher INSOL Ca content. The use of curd washing for cheeses made from high lactose milk prevented a large pH decrease during ripening; high rennet and draining pH values also retained more buffering constituents (i.e., INSOL Ca phosphate), which helped prevent a large pH decrease.     

Effect of different curd-washing methods on the insoluble Ca content and rheological properties of Colby cheese during ripening

A curd-washing step is used in the manufacture of Colby cheese to decrease the residual lactose content and, thereby, decrease the potential formation of excessive levels of lactic acid. The objective of this study was to investigate the effect of different washing methods on the Ca equilibrium and rheological properties of Colby cheese. Four different methods of curd-washing were performed. One method was batch washing (BW), where cold water (10°C) was added to the vat, with and without stirring, where curds were in contact with cold water for 5 min. The other method used was continuous washing (CW), with or without stirring, where curds were rinsed with continuously running cold water for approximately 7 min and water was allowed to drain immediately. Both methods used a similar volume of water. The manufacturing pH values were similar in all 4 treatments. The insoluble (INSOL) Ca content of cheese was measured by juice and acid-base titration methods and the rheological properties were measured by small amplitude oscillatory rheology. The levels of lactose in cheese at 1 d were significantly higher in CW cheese (0.06-0.11%) than in BW cheeses (∼0.02%). The levels of lactic acid at 2 and 12 wk were significantly higher in CW cheese than in BW cheeses. No differences in the total Ca content of cheeses were found. Cheese pH increased during ripening from approximately 5.1 to approximately 5.4. A decrease in INSOL Ca content of all cheeses during ripening occurred, although a steady increase in pH took place. The initial INSOL Ca content as a percent of total Ca in cheese ranged from 75 to 78% in all cheeses. The INSOL Ca content of cheese was significantly affected by washing method. Stirring during manufacturing did not have a significant effect on the INSOL Ca content of cheese during ripening. Batch-washed cheeses had significantly higher INSOL Ca contents than did CW cheeses during the first 4 wk of ripening. The maximum loss tangent values (meltability index) of CW cheese at 1 d and 1 wk were significantly higher compared with those of BW cheeses. In conclusion, different curd washing methods have a significant effect on the levels of lactose, lactic acid, meltability, and INSOL Ca content of Colby cheese during ripening.     

Calcium release from milk concentrated by ultrafiltration and diafiltration

The present work studied the solubilization of Ca during acidification in milk concentrated by ultrafiltration (UF) and diafiltration (DF). The effect of heating milk at 80°C for 15 min was also evaluated. In addition to measuring buffering capacity, the amount of Ca released as a function of pH was determined. The area of the maximum peak in buffering capacity observed at pH ~5.1, related to the presence of colloidal Ca phosphate, was significantly affected by casein volume fraction but did not increase proportionally with casein concentration. In addition, a lower buffering capacity and less solubilized Ca were measured in 2× DF milk compared with 2× UF milk. Heat treatment did not change the buffering capacity or Ca release in 1× and 2× concentrated milk. On the other hand, at a higher volume fraction (4×), more Ca was present in the soluble phase in heated 4× UF and DF milk compared with unheated milk. This is the first comprehensive study on the effect of concentration, distinguishing the effect of UF from that of DF, before and after heating, on Ca solubilization.     

Utilization of fourier transform infrared spectroscopy for measurement of organic phosphorus and bound calcium in cheddar cheese

The methods available for measuring organic P and bound Ca in cheese are either cumbersome or involve dilution of the cheese. Dilution of the cheese can lead to erroneous results, particularly in the case of bound Ca. Hence, the objective of this study was to evaluate the feasibility of Fourier transform infrared (FTIR) spectroscopy for direct measurement of organic P and bound Ca in Cheddar cheese. Two hundred sixteen samples of cheese were analyzed for protein-bound organic P, bound Ca using a water-extraction based method, and buffering curves. Additionally, the infrared spectra of the cheeses were collected between 4,000 and 650 cm⁻¹, at a resolution of 4 cm⁻¹, and 256 scans per sample. The spectral shifts in the infrared region from 1,050 to 900 cm⁻¹, in addition to the measured concentrations of organic P, bound Ca, and buffering peak area at pH 5.1, were used to develop calibration models using partial least squares (PLS) regression analysis. The spectral region of 956 to 946 cm⁻¹ correlated with the measured concentrations of organic P and the overall PLS model had a correlation (R²) of 0.76 between the predicted and measured concentrations. The spectral region at ∼980 cm⁻¹ was correlated with the measured concentrations of bound Ca, and the overall PLS model had a correlation (R²) of 0.70 between the predicted and measured concentrations. A similar spectral region at ∼980 cm⁻¹ was also correlated with the measured buffering peak areas and the overall PLS model had a correlation (R²) of 0.64 between the predicted and measured peak areas. A linear regression analysis between the bound Ca and buffering peak area demonstrated that bound Ca was correlated (R² = 0.73) with buffering peak area. This study demonstrates that FTIR can be used to measure organic P in cheeses. It also has the potential to be used for measuring bound Ca in undiluted cheeses, and for prediction of the buffering capacity of cheese.     

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

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

Effect of sodium hexametaphosphate concentration and cooking time on the physicochemical properties of pasteurized process cheese

Sodium hexametaphosphate (SHMP) is commonly used as an emulsifying salt (ES) in process cheese, although rarely as the sole ES. It appears that no published studies exist on the effect of SHMP concentration on the properties of process cheese when pH is kept constant; pH is well known to affect process cheese functionality. The detailed interactions between the added phosphate, casein (CN), and indigenous Ca phosphate are poorly understood. We studied the effect of the concentration of SHMP (0.25-2.75%) and holding time (0-20 min) on the textural and rheological properties of pasteurized process Cheddar cheese using a central composite rotatable design. All cheeses were adjusted to pH 5.6. The meltability of process cheese (as indicated by the decrease in loss tangent parameter from small amplitude oscillatory rheology, degree of flow, and melt area from the Schreiber test) decreased with an increase in the concentration of SHMP. Holding time also led to a slight reduction in meltability. Hardness of process cheese increased as the concentration of SHMP increased. Acid-base titration curves indicated that the buffering peak at pH 4.8, which is attributable to residual colloidal Ca phosphate, was shifted to lower pH values with increasing concentration of SHMP. The insoluble Ca and total and insoluble P contents increased as concentration of SHMP increased. The proportion of insoluble P as a percentage of total (indigenous and added) P decreased with an increase in ES concentration because of some of the (added) SHMP formed soluble salts. The results of this study suggest that SHMP chelated the residual colloidal Ca phosphate content and dispersed CN; the newly formed Ca-phosphate complex remained trapped within the process cheese matrix, probably by cross-linking CN. Increasing the concentration of SHMP helped to improve fat emulsification and CN dispersion during cooking, both of which probably helped to reinforce the structure of process cheese.     

Effect of trisodium citrate concentration and cooking time on the physicochemical properties of pasteurized process cheese

The effects of the concentration of trisodium citrate (TSC) emulsifying salt (0.25 to 2.75%) and holding time (0 to 20 min) on the textural, rheological, and microstructural properties of pasteurized process Cheddar cheese were studied using a central composite rotatable design. The loss tangent parameter (from small amplitude oscillatory rheology), extent of flow (derived from the University of Wisconsin Meltprofiler), and melt area (from the Schreiber test) all indicated that the meltability of process cheese decreased with increased concentration of TSC and that holding time led to a slight reduction in meltability. Hardness increased as the concentration of TSC increased. Fluorescence micrographs indicated that the size of fat droplets decreased with an increase in the concentration of TSC and with longer holding times. Acid-base titration curves indicated that the buffering peak at pH 4.8, which is due to residual colloidal calcium phosphate, decreased as the concentration of TSC increased. The soluble phosphate content increased as concentration of TSC increased. However, the insoluble Ca decreased with increasing concentration of TSC. The results of this study suggest that TSC chelated Ca from colloidal calcium phosphate and dispersed casein; the citrate-Ca complex remained trapped within the process cheese matrix. Increasing the concentration of TSC helped to improve fat emulsification and casein dispersion during cooking, both of which probably helped to reinforce the structure of process cheese.     

Effect of lactose standardization of milk using low-concentration factor ultrafiltration: Effect of reducing the lactose-to-casein ratio on the properties of milled-curd Cheddar cheese

The pH of cheese is determined by the amount of lactose fermented and the buffering capacity of the cheese. The buffering capacity of cheese is largely determined by the protein contents of milk and cheese and the amount of insoluble calcium phosphate in the curd, which is related to the rate of acidification. The objective of this study was to standardize both the lactose and casein contents of milk to better control final pH and prevent the development of excessive acidity in Cheddar cheese. This approach involved the use of low-concentration factor ultrafiltration of milk to increase the casein content (～5%), followed by the addition of water, ultrafiltration permeate, or both to the retentate to adjust the lactose content. We evaluated milks with 4 different lactose-to-casein ratios (L:CN): 1.8 (control milk), 1.4, 1.1, and 0.9. All cheesemilks had similar total casein (2.3%) and fat (3.4%) contents. These milks were used to make milled-curd Cheddar cheese, and we evaluated cheese composition, texture, functionality, and sensory properties over 9 mo of ripening. Cheeses made from milks with varying levels of L:CN had similar moisture, protein, fat, and salt contents, due to slight modifications during manufacture (i.e., cutting the gel at a smaller size than control) as well as control of acid development at critical steps (i.e., cutting the gel, whey drainage, salting). As expected, decreasing the L:CN led to cheeses with lower lactic acid, residual lactose, and insoluble Ca contents, as well as a substantial pH increase during cheese ripening in cheeses. The L:CN ratio had no significant effect on the levels of primary and secondary proteolysis. Texture profile analysis showed no significant differences in hardness values during ripening. Maximum loss tangent, an index of cheese meltability, was lower until 45 d for the L:CN 1.4 and 0.9 treatments, but after 45 d, all reduced L:CN cheeses had higher maximum loss tangent values than the control cheese (L:CN 1.8). Sensory analyses showed that cheeses made from milks with reduced L:CN contents had lower acidity, sourness, sulfury notes, and chewdown cohesiveness. Standardization of milk to a specific L:CN ratio, while maintaining a constant casein level in the milk, would allow Cheddar cheese manufacturers to have tighter control of pH and acidity.     

Influence of mixtures of calcium-chelating salts on the physicochemical properties of casein micelles

Calcium-chelating salts (CCS), such as phosphates and citrates, are often added to milk systems to modify physical properties like heat stability. The objective of this study was to investigate the effect of binary CCS mixtures on the properties of casein (CN) micelles including the distribution of Ca between the soluble and CN-bound states. Six binary CCS mixtures were prepared from 4 different types of CCS [i.e., trisodium citrate (TSC), disodium phosphate (DSP), tetrasodium pyrophosphate (TSPP), and sodium hexameta phosphate (SHMP)] by combining 2 CCS at a time in 5 different proportions (8.3:91.7, 29.2:70.8, 50:50, 70.8:29.2, and 91.7:8.3). Different concentrations of these mixtures (0, 0.1, 0.3, 0.5, and 0.7% wt/wt) were added to milk protein concentrate solutions (5% wt/wt) at pH 5.8. The ability of CCS to disperse CN particles and its interaction with Ca were assessed from turbidity measurements, acid-base titration behavior, and the quantity of CN-bound Ca and inorganic phosphate (Pi). Turbidity and the buffering peak at pH ∼5.0 during acid titration decreased with an increasing concentration of CCS. This was due to the chelation of Ca and the dispersion of CN micelles. The presence of TSC in mixtures decreased the amount of CN-bound Ca and Pi; however, the presence of TSPP in mixtures increased CN-bound Ca and Pi. When DSP was present at high proportions in mixtures of CCS, the CN-bound Ca and Pi slightly increased. When SHMP was used in mixtures of CCS, CN-bound Ca and Pi increased with the use of a low proportion of SHMP but decreased when SHMP was used at high proportions in the mixture. Combinations of DSP-TSPP used in the proportions 29.2:70.8, 50:50, and 70.8:29.2 resulted in the gelation of milk protein concentrates when the total CCS concentration was ≥0.3%. These results indicated that the type of CCS present in a mixture modified CN properties by various mechanisms, including chelation of Ca, dispersion of CN micelles, and formation of new types of Ca-CCS complexes. The type of interaction between the newly formed Ca-CCS complexes and the dispersed CN depended on the proportion, concentration, and type of CCS present in the mixtures. This information is useful in understanding how mixtures of CCS affect CN properties.     

Influence of calcium and phosphorus, lactose, and salt-to-moisture ratio on cheddar cheese quality: pH changes during ripening

The pH of cheese is an important attribute that influences its quality. Substantial changes in cheese pH are often observed during ripening. A combined effect of calcium, phosphorus, residual lactose, and salt-to-moisture ratio (S/M) of the cheese on the changes in cheese pH during ripening was investigated. Eight cheeses with 2 levels of Ca and P (0.67 and 0.47% vs. 0.53 and 0.39%, respectively), lactose at pressing (2.4 vs. 0.78%), and S/M (6.4 vs. 4.8%) were manufactured. All the cheeses were salted at a pH of 5.4, pressed for 5 h, and then ripened at 6 to 8°C. The pH of the salted curds before pressing and the cheeses during 48 wk of ripening was measured. Also, cheeses were analyzed for water-soluble Ca and P, organic P, and bound inorganic P during ripening. Changes in organic acids’ concentration and shifts in the distribution of Ca and P between different forms were studied in relation to changes in pH. Cheeses with low S/M exhibited a larger increase in acid production during ripening compared with high S/M cheeses. Cheeses with the highest concentration of bound inorganic P exhibited the highest pH, whereas cheeses with the lowest concentration of bound inorganic P exhibited the lowest pH among the 8 treatments. Although conversion of lactose to short-chain, water-soluble organic acids decreased cheese pH, bound inorganic phosphate buffered the changes in cheese pH. Production of acid in excess of the buffering capacity (which was the case in low Ca and P and low S/M treatments) led to a low pH, whereas solubilization of bound inorganic P in excess to acid production (which was the case in high Ca and P and high S/M treatments) led to an increase in pH. However, for cheeses with high Ca and P and low S/M, changes in cheese pH were influenced by the level of residual lactose. Hence, pH changes in Cheddar cheese can be modulated by a concomitant control on the amount and state of Ca and P, level of residual lactose, and S/M of the cheese.     

Influence of calcium and phosphorus, lactose, and salt-to-moisture ratio on Cheddar cheese quality: pH buffering properties of cheese

The pH buffering capacity of cheese is an important determinant of cheese pH. However, the effects of different constituents of cheese on its pH buffering capacity have not been fully clarified. The objective of this study was to characterize the chemical species and chemical equilibria that are responsible for the pH buffering properties of cheese. Eight cheeses with 2 levels of Ca and P (0.67 and 0.47% vs. 0.53 and 0.39%, respectively), residual lactose (2.4 vs. 0.78%), and salt-to-moisture ratio (6.4 vs. 4.8%) were manufactured. The pH-titration curves for these cheeses were obtained by titrating cheese:water (1:39 wt/wt) dispersions with 1 N HCl, and backtitrating with 1 N NaOH. To understand the role of different chemical equilibria and the respective chemical species in controlling the pH of cheese, pH buffering was modeled mathematically. The 36 chemical species that were found to be relevant for modeling can be classified as cations (Na⁺, Ca²⁺, Mg²⁺), anions (phosphate, citrate, lactate), protein-bound amino acids with a side-chain pKa in the range of 3 to 9 (glutamate, histidine, serine phosphate, aspartate), metal ion complexes (phosphate, citrate, and lactate complexes of Na⁺, Ca²⁺, and Mg²⁺), and calcium phosphate precipitates. A set of 36 corresponding equations was solved to give the concentrations of all chemical species as a function of pH, allowing the prediction of buffering curves. Changes in the calculated species concentrations allowed the identification of the chemical species and chemical equilibria that dominate the pH buffering properties of cheese in different pH ranges. The model indicates that pH buffering in the pH range from 4.5 to 5.5 is predominantly due to a precipitate of Ca and phosphate, and the protonation equilibrium involving the side chains of protein-bound glutamate. In the literature, the precipitate is often referred to as amorphous colloidal calcium phosphate. A comparison of experimental data and model predictions shows that the buffering properties of the precipitate can be explained, assuming that it consists of hydroxyapatite [Ca₅(OH)(PO₄)₃] or Ca₃(PO₄)₂. The pH buffering in the region from pH 3.5 to 4.5 is due to protonation of side-chain carboxylates of protein-bound glutamate, aspartate, and lactate, in order of decreasing significance. In addition, pH buffering between pH 5 to 8 in the backtitration results from the reprecipitation of calcium and phosphate either as CaHPO₄ or Ca₄H(PO₄)₃.     

Effect of natural cheese characteristics on process cheese properties

Natural cheese is the major ingredient utilized to manufacture process cheese. The objective of the present study was to evaluate the effect of natural cheese characteristics on the chemical and functional properties of process cheese. Three replicates of 8 natural (Cheddar) cheeses with 2 levels of calcium and phosphorus, residual lactose, and salt-to-moisture ratio (S/M) were manufactured. After 2 mo of ripening, each of the 8 natural cheeses was converted to 8 process cheese foods that were balanced for their composition, including moisture, fat, salt, and total protein. In addition to the standard compositional analysis (moisture, fat, salt, and total protein), the chemical properties (pH, total Ca, total P, and intact casein) and the functional properties [texture profile analysis (TPA), modified Schreiber melt test, dynamic stress rheometry, and rapid visco analysis] of the process cheese foods were determined. Natural cheese Ca and P, as well as S/M, significantly increased total Ca and P, pH, and intact casein in the process cheese food. Natural cheese Ca and P and S/M also significantly affected the final functional properties of the process cheese food. With the increase in natural cheese Ca and P and S/M, there was a significant increase in the TPA-hardness and the viscous properties of process cheese food, whereas the meltability of the process cheese food significantly decreased. Consequently, natural cheese characteristics such as Ca and P and S/M have a significant influence on the chemical and the final functional properties of process cheese.     

Effect of camel chymosin on the texture,functionality, and sensory properties of low-moisture,part-skim Mozzarella cheese

The objective of this study was to compare the effect of coagulant (bovine calf chymosin, BCC, or camel chymosin, CC), on the functional and sensory properties and performance shelf-life of low-moisture, part-skim (LMPS) Mozzarella. Both chymosins were used at 2 levels [0.05 and 0.037 international milk clotting units (IMCU)/mL], and clotting temperature was varied to achieve similar gelation times for each treatment (as this also affects cheese properties). Functionality was assessed at various cheese ages using dynamic low-amplitude oscillatory rheology and performance of baked cheese on pizza. Cheese composition was not significantly different between treatments. The level of total calcium or insoluble (INSOL) calcium did not differ significantly among the cheeses initially or during ripening. Proteolysis in cheese made with BCC was higher than in cheeses made with CC. At 84 d of ripening, maximum loss tangent values were not significantly different in the cheeses, suggesting that these cheeses had similar melt characteristics. After 14 d of cheese ripening, the crossover temperature (loss tangent = 1 or melting temperature) was higher when CC was used as coagulant. This was due to lower proteolysis in the CC cheeses compared with those made with BCC because the pH and INSOL calcium levels were similar in all cheeses. Cheeses made with CC maintained higher hardness values over 84 d of ripening compared with BCC and maintained higher sensory firmness values and adhesiveness of mass scores during ripening. When melted on pizzas, cheese made with CC had lower blister quantity and the cheeses were firmer and chewier. Because the 2 types of cheeses had similar moisture contents, pH values, and INSOL Ca levels, differences in proteolysis were responsible for the firmer and chewier texture of CC cheeses. When cheese performance on baked pizza was analyzed, properties such as blister quantity, strand thickness, hardness, and chewiness were maintained for a longer ripening time than cheeses made with BCC, indicating that use of CC could help to extend the performance shelf-life of LMPS Mozzarella.     

Effects of the concentration of insoluble calcium phosphate associated with casein micelles on the functionality of directly acidified cheese

Directly acidified cheeses with different insoluble Ca (INS Ca) contents were made to test the hypothesis that the removal of INS Ca from casein micelles (CM) would directly contribute to the softening and flow behavior of cheese at high temperature. Skim milk was directly acidified with dilute lactic acid to pH values of 6.0, 5.8, 5.6, or 5.4 to remove INS Ca (pH trial). Lowering milk pH also reduced protein charge repulsion, which could influence melt. In a second treatment, EDTA (0, 2, 4, or 6 mM) was added to skim milk that was subsequently acidified to pH 6.0 (EDTA trial). Both types of milks were then made into directly acidified cheese. Cheese properties were determined at approximately 10 h after pressing to reduce possible confounding effects of proteolysis. The INS Ca content was determined by the acid-base titration method. Dynamic low-amplitude oscillatory rheology was used to measure the viscoelastic properties of cheese during heating from 5 to 80°C. The composition of all cheeses was as similar as possible, with cheese-making procedures being modified to obtain similar moisture contents (∼55%). Insoluble Ca contents of cheeses significantly decreased with a reduction in pH or with the addition of EDTA to skim milk. The pH values of cheeses in the pH trial varied, but all cheeses in the EDTA trial had similar pH values (∼5.73). In the pH trial, the reduction in cheese pH and consequent decrease in INS Ca content resulted in a reduction in the G′ values of cheeses at 20°C. In contrast, the G′ values at 20°C in cheeses from the EDTA trial increased with EDTA addition up to 4 mM EDTA. The G′ values at 70°C of cheeses from the pH trial decreased with a decrease in cheese pH, and a similar decrease was observed in the G′ values of cheese from the EDTA trial with an increase in EDTA concentration even though these cheeses had a similar pH value. In both trials, loss tangent (LT) values increased with temperatures >30°C and reached a maximum at approximately 70°C. In the pH trial, LT values at 70°C increased from 1.50 to 4.24 with a decrease in cheese pH from 5.78 to 5.21. The LT values increased from 1.43 to 3.23 with an increase in the concentration of added EDTA from 0 to 6 mM. In the EDTA trial, the decrease in G′ and increase in LT values at 70°C were due to the reduction in INS Ca content, because the pH values of these cheeses were the same. It can be concluded that the loss of INS Ca increases the melting in cheeses that have the same pH and gross chemical composition, and removal of INS Ca can even make cheese at high pH (∼5.73) exhibit reasonable melt characteristics.     

Effect of salt addition on the micellar composition of milk subjected to pH reversible CO2 acidification

Response surface methodology was used to investigate the effect of salt supplementation on the micellar composition of reconstituted skim milk subjected to acidification by CO2 pressure to pH 5.8, followed by depressurization under vacuum. Using a Doehlert design, calcium and phosphate were added to skim milk in the range of 0 to 25 mmol/kg and 0 to 16 mmol/kg of milk, respectively, and the pH was adjusted to 6.65 +/- 0.02. After carbonation, the milk sample was depressurized, and the pH returned to its initial value without modification of the ionic strength. Micellar composition was assessed by the concentration of micellar Ca, P, Mg, and protein, and the buffering properties of milk. The second order polynomial models satisfactorily predicted the effect of salt supplementation on the micellar composition (R2adj > 0.75). Added calcium was the most determinant factor, and favored the removal of Ca, P, Mg, and proteins from the soluble phase to the micellar phase when this addition was less than 17.5 mmol/kg of milk. Above this concentration, only the concentration of micellar Ca increased. The buffering response surface showed that the amount of micellar calcium phosphate increased to a maximum upon addition of 17.5 mmol of Ca/kg. By comparison with a control sample (supplemented but untreated skim milk), changes were essentially due to salt supplementation and not to the CO2 treatment. We suggest that Ca formed micellar calcium phosphate when added at a concentration less than 17.5 mmol/kg; whereas above this concentration, Ca bound directly to micellar proteins.     

Invited review: perspectives on the basis of the rheology and texture properties of cheese

Physical and chemical properties of cheese, such as texture, color, melt, and stretch, are primarily determined by the interaction of casein (CN) molecules. This review will discuss CN chemistry, how it is influenced by the cheese-making process, and how it impinges on the final product, cheese. We attempt to demonstrate that the application of principles governing the molecular interactions of CN can be useful in understanding the many physical and chemical properties of cheese and, in turn, how this can be used by the cheesemaker to produce the desired cheese. The physical properties of cheese (as well as flavor) are influenced by a number of factors including: milk composition; milk quality; temperature; the rate and extent of acidification by the starter bacteria; the pH history of cheese; the concentration of Ca salts (proportions of soluble and insoluble forms); extent and type of proteolysis, and other ripening reactions. Our hypothesis is that these factors also control and modify the nature and strength of CN interactions. The approach behind the recently proposed dual-binding model for the structure and stability of CN micelles is used as a framework to understand the physical and chemical properties of cheese.     

The effects of κ-casein polymorphism on the texture and functional properties of mozzarella cheese

The present study compared the texture and functional properties of mozzarella cheese made with milk containing different of genetic polymorphisms κ-casein (AA, AB, AE or BE). The genotype of κ-casein in the milk from individual Holstein cow was determined by pyrosequencing method. Full-fat Mozzarella cheese was made from pooled milk from 3 cows with the same κ-casein genotype and analysed 7 d after manufacture. The cheese made from type AB contained the highest level of fat and Ca/protein, and the lowest moisture content. The cheese made from type AB milk was harder and chewier than cheese made from type AE and BE milk. The cheese made from type AB and AA milk had higher stretchability but lower meltability and flowability than type AE and BE. In summary, the cheese made from type AB milk had different texture and functionality quality than that made from type AE or BE.     

A model system for studying the effects of colloidal calcium phosphate concentration on the rheological properties of Cheddar cheese

A novel model system was developed for studying the effects of colloidal Ca phosphate (CCP) concentration on the rheological properties of Cheddar cheese, independent of proteolysis and any gross compositional variation. Cheddar cheese slices (disks; diameter = 50 mm, thickness = 2 mm) were incubated in synthetic Cheddar cheese aqueous phase solutions for 6 h at 22°C. Control (unincubated) Cheddar cheese had a total Ca and CCP concentration of 2.80 g/100 g of protein and 1.84 g of Ca/100 g of protein, respectively. Increasing the concentration of Ca in the synthetic Cheddar cheese aqueous phase solution incrementally in the range from 1.39 to 8.34 g/L significantly increased the total Ca and CCP concentration of the cheese samples from 2.21 to 4.59 g/100 g of protein and from 1.36 to 2.36 g of Ca/100 g of protein, respectively. Values of storage modulus (index of stiffness) at 70°C increased significantly with increasing concentrations of CCP, but the opposite trend was apparent at 20°C. The maximum in loss tangent (index of meltability/flowability) decreased significantly with increasing concentration of CCP, and there was no significant effect on the temperature at which the maximum in loss tangent occurred (68 to 70°C). Fourier transform mechanical spectroscopy showed the frequency dependence of all of the cheese samples increased with increasing temperature; however, solubilization of CCP increased the frequency dependence of the cheese matrix only in the high temperature region (i.e., >35°C). These results support earlier studies that hypothesized that the concentration of CCP strongly modulates the rheological properties of cheese.     

Effect of high-pressure treatment of ewe raw milk curd at 200 and 300 MPa on characteristics of Hispánico cheese

Hispánico cheese is a semihard variety made from a mixture of cow and ewe milks. Production of ewe milk declines in summer and autumn. To surmount the seasonal shortage of ewe milk and prevent the inactivation of milk enzymes by pasteurization, curd made in spring from ewe raw milk was pressurized at 200 and 300 MPa and stored frozen for 4 mo. Thawed ewe milk curds were added to fresh curd made from pasteurized cow milk for the manufacture of experimental Hispánico cheeses. Control cheese was made from a mixture of pasteurized cow and ewe milk in the same proportions as those used for experimental cheeses. Experimental cheeses exhibited lower dry matter content, higher aminopeptidase activity and total free amino acid concentration, and higher levels of acetic and propionic acids, aldehydes, alcohols, and esters compared with control cheese. In contrast, the concentration of total free fatty acids and ketones and the levels of textural parameters were significantly higher in control cheese. The use of ewe raw milk curd pressurized at 200 and 300 MPa, stored frozen and thawed for Hispánico cheese manufacture, was generally beneficial for cheese characteristics and increased cheese yield because of the lower dry matter content of experimental cheeses.