Heat stability of milk: pH-dependent dissociation of micellar κ-casein on heating milk at ultra high temperatures

1985 ◽  
Vol 52 (4) ◽  
pp. 529-538 ◽  
Author(s):  
Harjinder Singh ◽  
Partick F. Fox
Keyword(s):  
Whey Protein ◽  
Whey Proteins ◽  
Heat Stability ◽  
Casein Micelle ◽  
Casein Micelles ◽  
Ph Profile ◽  
Ph Values ◽  
Maximum Stability ◽  
Β Lactoglobulin ◽  
Maximum Minimum

SUMMARYPreheating milk at 140 °C for 1 min at pH 6·6, 6·8, 7·0 or 7·2 shifted the heat coagulation time (HCT)/pH profile to acidic values without significantly affecting the maximum stability. Whey proteins (both β-lactoglobulin and α-lactalbumin) co-sedimented with the casein micelles after heating milk at pH < 6·9 and the whey protein-coated micelles, dispersed in milk ultrafiltrate, showed characteristic maxima–minima in their HCT/pH profile. Heating milk at higher pH values (> 6·9) resulted in the dissociation of whey proteins and κ-casein-rich protein from the micelles and the residual micelles were unstable, without a maximum–minimum in the HCT/pH profile. Preformed whey protein–casein micelle complexes formed by preheating (140 °C for 1 min) milk at pH 6·7 dissociated from the micelles on reheating (140 °C for 1 min) at pH > 6·9. The dissociation of micellar-κ-casein, perhaps complexed with whey proteins, may reduce the micellar zeta potential at pH ≃ 6·9 sufficiently to cause a minimum in the HCT/pH profile of milk.

Heat stability of milk: influence of modifying sulphydryl-disulphide interactions on the heat coagulation time–pH profile

1987 ◽  
Vol 54 (3) ◽  
pp. 347-359 ◽  
Author(s):  
Harjinder Singh ◽  
Patrick F. Fox
Keyword(s):  
Heat Stability ◽  
Casein Micelle ◽  
Coagulation Time ◽  
Casein Micelles ◽  
Ph Profile ◽  
Ph Values ◽  
Stabilizing Effect ◽  
Ph Range ◽  
Β Lactoglobulin ◽  
Minimum Ratio

SummaryAddition of reducing agents such as 2-mercaptoethanol (2-ME), dithio-threitol and Na sulphite to milk markedly reduced its heat stability at pH values below 7·1. 2-ME reversibly destabilized milk or serum protein-free casein micelle dispersions and promoted the release of κ-casein-rich protein from the micelles. Reduction of either casein micelles or β-lactoglobulin (β-lg) with 2-ME and subsequent blocking of the newly formed –SH groups with N-ethylmaleimide irreversibly reduced the maximum to minimum ratio in the heat stability profile. 2-ME disrupted κ-casein/β-lg complexes and stripped κ-casein from the micelles on heating. The milk or caseinate systems were thus destabilized. Addition of KBrO4 or iodosobenzoate to milk at 5 HIM eliminated the minimum but destabilized milk in the region of the maximum. However, KIO3 at 5 mm had a strong stabilizing effect throughout the pH range 6·5–7·3.

Heat stability of milk: role of β-lactoglobulin in the pH-dependent dissociation of micellar κ-casein

1987 ◽  
Vol 54 (4) ◽  
pp. 509-521 ◽  
Author(s):  
Harjinder Singh ◽  
Patrick F. Fox
Keyword(s):  
Heat Stability ◽  
Casein Micelle ◽  
Casein Micelles ◽  
Degree Of Hydration ◽  
Ph Values ◽  
High Concentrations ◽  
Ph Range ◽  
Β Lactoglobulin ◽  
Micelle Hydration

SummaryOn heating casein micelle systems containing β-lactoglobulin (β-lg) at 90°C for 10 min, β-lg complexed with casein micelles at pH < 6·9, probably as a result of interaction with κ-casein via sulphydryl-disulphide interchange, and co-sedimented with the micelles on ultracentrifugation. Complex formation with β-lg appeared to prevent the dissociation of micellar κ-casein on heating. However, at pH ≥ 6·9, κ-casein/β-lg complexes dissociated from the micelles on heating, thus enhancing the release of micellar κ-casein. High concentrations of β-lg (≥0·8%) induced coagulation at pH 7·3, essentially by promoting the dissociation of micellar κ-casein. It appeared that αs1-, αs2-, β- and κ-caseins dissociated from serum protein-free casein micelles to equal extents, but the presence of β-lg specifically enhanced the dissociation of κ-casein at pH values ≥ 6·9. Micelle hydration increased slightly when casein micelles were heated in the presence of β-lg at pH 6·7, while at pH 7·3 β-lg decreased the degree of hydration of casein micelles. Formation of a complex between β-lg and κ-casein appeared to stabilize the micelles in the pH range 6·5–6·7, possibly via increased micellar charge or degree of hydration or by preventing the dissociation of κ-casein.

Heat stability of milk: influence of κ-casein hydrolysis

1978 ◽  
Vol 45 (2) ◽  
pp. 173-181 ◽  
Author(s):  
Patrick F. Fox ◽  
Catherine M. Hearn
Keyword(s):  
Heat Stability ◽  
Principal Factor ◽  
Coagulation Time ◽  
Casein Micelles ◽  
Ph Profile ◽  
Acetylneuraminic Acid ◽  
Ph Values ◽  
Colloidal Calcium Phosphate ◽  
Maximum Stability ◽  
Ph Range

SummaryThe release of 12% (w/v) TCA-soluble N-acetylneuraminic acid from casein at 5 temperatures between 110 and 150°C was determined and showed a Q10 °C about 3; coagulation occurred when about 20% of the κ-casein was hydrolysed. Renneting of milk or colloidal calcium phosphate-free milk rendered the caseinate very heat-labile at its normal pH when more than about 20% of the κ-casein had been hydrolysed. Heat stability at the pH of maximum stability was not significantly decreased until after very prolonged renneting, but stability at pH values alkaline to the minimum was very sensitive to such hydrolysis suggesting that κ-casein is the principal factor responsible for heat stability in that region. Systems which do not have a maximum or minimum in the heat coagulation time–pH profile, i.e. serum protein-free casein micelles in milk diffusate, or milk which had been dialysed against water, were destabilized by renneting throughout the pH range 6·4–7·4.

High-pressure treatment of milk: effects on casein micelle structure and on enzymic coagulation

2000 ◽  
Vol 67 (1) ◽  
pp. 31-42 ◽  
Author(s):  
ERIC C. NEEDS ◽  
ROBERT A. STENNING ◽  
ALISON L. GILL ◽  
VICTORIA FERRAGUT ◽  
GILLIAN T. RICH
Keyword(s):  
Whey Proteins ◽  
Skim Milk ◽  
Casein Micelle ◽  
Casein Micelles ◽  
Dense Networks ◽  
Protein Levels ◽  
Micelle Structure ◽  
Milk Casein ◽  
Β Lactoglobulin ◽  
Gel Network

High isostatic pressures up to 600 MPa were applied to samples of skim milk before addition of rennet and preparation of cheese curds. Electron microscopy revealed the structure of rennet gels produced from pressure-treated milks. These contained dense networks of fine strands, which were continuous over much bigger distances than in gels produced from untreated milk, where the strands were coarser with large interstitial spaces. Alterations in gel network structure gave rise to differences in rheology with much higher values for the storage moduli in the pressure-treated milk gels. The rate of gel formation and the water retention within the gel matrix were also affected by the processing of the milk. Casein micelles were disrupted by pressure and disruption appeared to be complete at treatments of 400 MPa and above. Whey proteins, particularly β-lactoglobulin, were progressively denatured as increasing pressure was applied, and the denatured β-lactoglobulin was incorporated into the rennet gels. Pressure-treated micelles were coagulated rapidly by rennet, but the presence of denatured β-lactoglobulin interfered with the secondary aggregation phase and reduced the overall rate of coagulation. Syneresis from the curds was significantly reduced following treatment of the milk at 600 MPa, probably owing to the effects of a finer gel network and increased inclusion of whey protein. Levels of syneresis were more similar to control samples when the milk was treated at 400 MPa or less.

Heat stability of milk: influence of colloidal calcium phosphate and β-lactoglobulin

1975 ◽  
Vol 42 (3) ◽  
pp. 427-435 ◽  
Author(s):  
P. F. Fox ◽  
M. C. T. Hoynes
Keyword(s):  
Calcium Phosphate ◽  
Fold Increase ◽  
Heat Stability ◽  
Protein Charge ◽  
Ph Values ◽  
Colloidal Calcium Phosphate ◽  
Maximum Stability ◽  
Ph Range ◽  
Ph Curve ◽  
Β Lactoglobulin

SummaryReduction of the level of colloidal calcium phosphate (CCP) progressively increased the heat stability of milk at pH values <~7·0 and increased the pH of maximum stability. Removal of 40% CCP also stabilized the system at the pH of minimum stability, but removal of ≥60% CCP rendered milk very unstable at pH values >7·2, an effect not offset by a 4-fold increase in κ-casein concentration. Doubling CCP had a slight destabilizing effect in the pH range 6·5–7·5.Addition of β-lactoglobulin to serum protein-free casein micelles had a marked destabilizing effect at pH values > ~6·8, but increased stability in the pH range 6·4–6·8. β-Lactoglobulin had a similar and more apparent effect on the heat stability of Na caseinate dissolved in milk diffusate.It is suggested that rather than being a stabilizing factor responsible for the maximum in the heat stability-pH curve, the true effect of β-lactoglobulin is to shift the curve to more acid pH values (reason unknown) and to sensitize the caseinate system to heat-induced Ca phosphate precipitation at pH values > ~7·0. Low stability at ~pH 7·0 introduces an apparent maximum in the heat stability-pH curve at ~pH 6·8, but this has no independent existence. At pH values >7·2, increased protein charge more than off-sets the influence of heat-precipitated CCP and stability again increases in micellar but not in soluble casein systems.

Heat stability of milk: further studies on the pH-dependent dissociation of micellar κ-casein

1986 ◽  
Vol 53 (2) ◽  
pp. 237-248 ◽  
Author(s):  
Harjinder Singh ◽  
Patrick F. Fox
Keyword(s):  
Whey Protein ◽  
Cetyltrimethylammonium Bromide ◽  
Opposite Effect ◽  
Whey Proteins ◽  
Sodium Dodecyl ◽  
Skim Milk ◽  
Heat Stability ◽  
Casein Micelles ◽  
Acetylneuraminic Acid ◽  
Ph Dependent

SUMMARYWhey protein complexed and became co-sedimentable with casein micelles after heating milk at ≥ 90°C for 10 min at pH ≤ 6·9 while at higher pH values (7·3) whey proteins and κ-casein-rich protein dissociated from the micelles on heating. κ-Casein-deficient micelles were more sensitive to heat, Ca2+ or ethanol than whey protein-coated or native micelles and were readily coagulable by rennet. Isolated κ-casein added to skim milk before preheating (90°C for 10 min) did not associate with the micelles at pH ≥ 6·9. Sodium dodecyl sulphate increased the level of both non-sedimentable N (NSN) and N-acetylneuraminic acid (NANA) and shifted the NSN-pH and NANA-pH curves to more acidic values while cetyltrimethylammonium bromide had the opposite effect. It is suggested that the pH-dependent dissociation in micellar κ-casein, which appears to be reversible, depends on the surface charge on the micelles; at a certain negative charge, disruption of hydrophobic and electrostatic forces could result in the dissociation of κ-casein from the casein micelles.

Influence of 2-mercaptoethanol on heat stability of concentrated whey-protein-free milk and formation of soluble casein

1984 ◽  
Vol 51 (3) ◽  
pp. 439-445 ◽  
Author(s):  
Takayoshi Aoki ◽  
Yoshitaka Kako
Keyword(s):  
Whey Protein ◽  
Heat Stability ◽  
Coagulation Time ◽  
Casein Micelles ◽  
Ph Profile ◽  
Very High

SummaryThe heat coagulation time (HCT) of concentrated whey-protein-free (WPF) milk measured at 120 and 130 °C was reduced to by addition of 5–20 mM-2-mercaptoethanol (ME). However, although the amount of soluble casein formed on heating was doubled by addition of ME, the shape of the HCT–pH profile was affected only slightly. The proportion of κ-casein in the soluble casein from heated concentrated WPF milk containing ME was very high, though it was somewhat lower than that of the soluble casein from heated concentrated WPF milk containing no ME. No solubilization of colloidal Ca phosphate was observed in either unheated or heated concentrated WPF milk on addition of ME. These facts suggest that ME probably promotes the formation of soluble casein with release of κ-casein from micelles on heating, thus destabilizing the casein micelles.

High pressure treatment of bovine milk: effects on casein micelles and whey proteins

2004 ◽  
Vol 71 (1) ◽  
pp. 97-106 ◽  
Author(s):  
Thom Huppertz ◽  
Patrick F Fox ◽  
Alan L Kelly
Keyword(s):  
High Pressure ◽  
Treatment Time ◽  
Whey Proteins ◽  
Bovine Milk ◽  
Casein Micelle ◽  
Pressure Treatment ◽  
Casein Micelles ◽  
High Pressure Treatment ◽  
Micelle Size ◽  
Β Lactoglobulin

Effects of high pressure (HP) on average casein micelle size and denaturation of α-lactalbumin (α-la) and β-lactoglobulin (β-lg) in raw skim bovine milk were studied over a range of conditions. Micelle size was not influenced by treatment at pressures <200 MPa, but treatment at 250 MPa increased micelle size by ∼25%, while treatment at [ges ]300 MPa irreversibly reduced it to ∼50% of that in untreated milk. The increase in micelle size after treatment at 250 MPa was greater with increasing treatment time and temperature and milk pH. Treatment times [ges ]2 min at 400 MPa resulted in similar levels of micelle disruption, but increasing milk pH to 7·0 partially stabilised micelles against HP-induced disruption. Denaturation of α-la did not occur [les ]400 MPa, whereas β-lg was denatured at pressures >100 MPa. Denaturation of α-la and β-lg increased with increasing pressure, treatment time and temperature and milk pH. The majority of denatured β-lg was apparently associated with casein micelles. These effects of HP on casein micelles and whey proteins in milk may have significant implications for properties of products made from HP-treated milk.

Disruption and reassociation of casein micelles during high pressure treatment: influence of whey proteins

2007 ◽  
Vol 74 (2) ◽  
pp. 194-197 ◽  
Author(s):  
Thom Huppertz ◽  
Cornelis G de Kruif
Keyword(s):  
High Pressure ◽  
Treatment Time ◽  
Light Transmission ◽  
Whey Proteins ◽  
Casein Micelle ◽  
Casein Micelles ◽  
Β Lactoglobulin ◽  
Steric Stabilisation ◽  
Prolonged Holding

In the study presented in this article, the influence of added α-lactalbumin and β-lactoglobulin on the changes that occur in casein micelles at 250 and 300 MPa were investigated by in-situ measurement of light transmission. Light transmission of a serum protein-free casein micelle suspension initially increased with increasing treatment time, indicating disruption of micelles, but prolonged holding of micelles at high pressure partially reversed HP-induced increases in light transmission, suggesting reformation of micellar particles of colloidal dimensions. The presence of α-la and/or β-lg did not influence the rate and extent of micellar disruption and the rate and extent of reformation of casein particles. These data indicate that reformation of casein particles during prolonged HP treatment occurs as a result of a solvent-mediated association of the micellar fragments. During the final stages of reformation, κ-casein, with or without denatured whey proteins attached, associates on the surface of the reformed particle to provide steric stabilisation.

Heat stability of milk: influence of denaturable proteins and detergents on pH sensitivity

1978 ◽  
Vol 45 (2) ◽  
pp. 159-172 ◽  
Author(s):  
Patrick F. Fox ◽  
Catherine M. Hearn
Keyword(s):  
Tween 80 ◽  
Heat Stability ◽  
Ph Sensitivity ◽  
Ammonium Bromide ◽  
Casein Micelle ◽  
Maximum Stability ◽  
Ph Range ◽  
Triton X ◽  
Ph Curve ◽  
Β Lactoglobulin

Summaryα-Lactalbumin and SDS in addition to β-lactoglobulin introduced pH sensitivity to the heat stability–pH curve of serum protein free casein micelles particularly by increasing stability in the pH range 6·4–6·7. Bovine serum albumin, ovalbumin and lysozyme caused marked destabilization of milk and casein micelle suspensions throughout the pH range 6·4–7·4. Tetramethyl ammonium bromide caused destabilization of milk at pH values > 7·0, but had no effect in the region of maximum stability while the non-ionic detergents Triton X-100 and Tween 80 had no effect on heat stability.

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