Casein micelle stabilization

Hydrolysis of the casein micelle-stabilizing K-casein by the action of selected acid proteinases (rennets), and the resultant slow quiescent aggregation of the destabilized micelles in the presence of calcium ions ( 3 mM) at 30-36°C (e.g., for most rennet-curd cheeses such as Cheddar, Mozzarella and Gouda)... 

Horne, D. S. (2009). Casein micelles structure and stability. In "Milk Proteins From Expression to Food", (A. Thompson, M. Boland, and H. Singh, Eds), pp. 133-179. Academic Press, San Diego. 

Spagnuolo, P. A., Dalgleish, D. G., Goff, H. D., and Morris, E. R. (2005). Kappa-carrageenan interactions in systems containing casein micelles and polysaccharide stabilizers. Food Hydrocolloids 19,371-377. 

The elastic modulus (G ) of MP, BCAS, and BLG5 rapidly rose to plateaus that corresponded to different G saturations (Gjat) (Table 2). MP and BCAS coagula showed the more important Gsat value (142 N/m ), meaning that the emulsions stabilized by skim milk proteins (mainly casein micelles) and 6-casein formed the coagula with the strongest protein network. 

The following factors must be considered when assessing the stability of the casein micelle The role of Ca++ is very significant in milk. More than 90% of the calcium content of skim milk is associated in some way or another with the casein micelle. The removal of Ca++ leads to reversible dissociation of P-casein without micellular disintegration. The addition of Ca++ leads to aggregation. The same reaction occurs between the individual caseins in the micelle, but not as much because there is no secondary structure in casein proteins. 

Van der Waals forces There has been some success in relating these forces to micellar stability. However, the steric stabilization has been found to be also of some importance. Especially, the hairy layer interferes with the interparticle approach. There are several factors that will affect the stability of the casein micelle system ... 

The weak physical forces that hold together self-assembled nanoparticles are, of course, susceptible to disruption under the influence of thermodynamic and/or mechanical stresses. Hence some workers have investigated ways to reinforce nanoscale structures via covalent bonding. For instance, improved stability of protein nanoparticles, in particular, casein micelles, can be achieved by enzymatic cross-linking with the enzyme transglutaminase, which forms bonds between protein-bound glutamine and lysine residues. By this means native casein micelles can be converted from semi-reversible association colloids into permanent nanogel particles (Huppertz and de Kruif, 2008). 

Huppertz, T., de Kruif C.G. (2008). Structure and stability of nanogel particles prepared by internal cross-linking of casein micelles. International Dairy Journal, 18, 556-565. 

The casein responsible for colloidal stabilization of the casein micelle is K-casein. This glycoprotein has a molecular weight of 19 kDa and is composed of 169 amino acids (Swaisgood, 2003). K-Casein is special in being the only casein component that is insensitive to calcium ions (up to concentrations of 400 mM). 

The presence of casein-polysaccharide interactions is commonly invoked to explain the mechanistic stabilizing role of food hydrocolloids in dairy colloids (Dickinson, 1998a). Thus, for example, under conditions where the casein micelles and K-carrageenan both carry a net negative... 

Experiments on interactions of polysaccharides with casein micelles show similar trends to those with casein-coated droplets. For example, Maroziene and de Kruif (2000) demonstrated the pH-reversible adsorption of pectin molecules onto casein micelles at pH = 5.3, with bridging flocculation of casein micelles observed at low polysaccharide concentrations. In turn, Tromp et al. (2004) have found that complexes of casein micelles with adsorbed high-methoxy pectin (DE = 72.2%) form a self-supporting network which can provide colloidal stability in acidified milk drinks. It was inferred that non-adsorbed pectin in the serum was linked to this network owing to the absence of mobility of all the pectin in the micellar casein dispersion. Hence it seems that the presence of non-adsorbed pectin is not needed to maintain stability of an acid milk drink system. It was stated by Tromp et al. (2004) that the adsorption of pectin was irreversible in practical terms, i.e., the polysaccharide did not desorb under the influence of thermal motion. 

Principal micelle characteristics. The structure of the casein micelles has attracted the attention of scientists for a considerable time. Knowledge of micelle structure is important because the stability and behaviour of the micelles are central to many dairy processing operations, e.g. cheese manufacture, stability of sterilized, sweetened-condensed and reconstituted milks and frozen products. Without knowledge of the structure and properties of the casein micelle, attempts to solve many technological problems faced by the dairy industry will be empirical and not generally applicable. From the academic viewpoint, the casein micelle presents an interesting and complex problem in protein quaternary structure. 

Although CCP represents only about 6% of the dry weight of the casein micelle, it plays an essential role in its structure and properties and hence has major effects on the properties of milk it is the integrating factor in the casein micelle without it, milk is not coagulable by rennet and its heat and calcium stability properties are significantly altered. In fact, milk would be a totally different fluid without colloidal calcium phosphate. 

Figure 9.19 Effect of pH on the heat stability of type A milk (A), type B milk ( ) and whey protein-free casein micelle dispersions (O) (from Fox, 1982).

All the heat-induced changes discussed would be expected to cause major alterations in the casein micelles, but the most significant change with respect to heat coagulation appears to be the decrease in pH - if the pH is readjusted occasionally to pH 6.7, milk can be heated for several hours at 140°C without coagulation. The stabilizing effect of urea is, at least partially,... 

As discussed in Chapter 4, the casein micelles are stabilized by tc-casein, which represents 12-15% of the total casein and is located mainly on the surface of the micelles such that its hydrophobic N-terminal region reacts hydrophobically with the calcium-sensitive asl-, as2- and j8-caseins while its hydrophilic C-terminal region protrudes into the surrounding aqueous environment, stabilizing the micelles by a negative surface charge and steric stabilization. 

Hill, R. J. and Wake, R. G. 1969. Amphiphile nature of -casein as the basis for its micelle stabilizing properties. Nature 221, 635-639. 

The binding of milk lipases to casein micelles apparently imparts some stability to the enzyme, for as purification progresses, the milk lipase becomes less stable, and more so as the concentration of casein decreases (Downey and Andrews 1966 Egelrud and Olivecrona 1972). 

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