Casein micelle precipitation

Destruction of the casein micelles in the milk with subsequent precipitation of the casein can be accomplished in a number of ways. The action of heat or the action of alcohols, acids, salts and the enzyme rennet all bring about precipitation. In commercial practise the two techniques used employ either acid coagulation or rennet coagulation mechanisms. 

Co-preclpltate is an insoluble milk protein product that is produced by heating skinimllk to high temperatures ( > 90 C) to denature the whey proteins and complex them with the casein micelles. The heated system is subsequently adjusted to isoelectric point conditions of pH 4.5-5 to precipitate the complexed whey protein-casein micelles, centrifuged or filtered to recover the precipitate, washed and dryed. The resulting product, which is virtually insoluble, exhibits only minor functionality in most typical emulsification applications. 

Although the gelation properties of whey proteins are of great importance in many foods (Mulvihill, 1992) and it is possible to form a weak gel in creams by the formation of a continuous network of fat globules, most important milk gels are those involving casein micelles which can be made to form a gel matrix either by isoelectric precipitation (acid-induced gel) or by the action of a proteolytic enzyme (rennet-induced gel). Both gel types... 

Lowering the pH of milk to 4.6 solubilizes colloidal calcium phosphate. This removes its neutralizing effect, allowing electrostatic interactions between micelles. Under these conditions, micelles coagulate and precipitate from solution. Kudo (1980C) showed that release of whey proteins and K-casein from casein micelle surfaces as the pH is increased from 6.2 to 7.2 allows micelles to stick together and precipitate from solution. 

Stability of the complex protein system of milk or whey is decreased by concentration (Fox 1982 Muir and Sweetsur 1978 Sweetsur and Muir 1980B). In addition to closer packing of casein micelles and other proteins in concentrated milk, calcium phosphate is precipitated so that the pH decreases (Fox 1982). The pH effect causes protein which would be soluble at a normal solids concentration to precipitate. Casein in milk concentrated to three times its original solids level forms a flocculent after 1 to 3 weeks at -8°C (Lonergan 1978). 

According to Bloomfield and Mead (1974), The ultimate goal of all workers on casein is to reconstitute micelles with native properties from the separated constituents of skim-milk. This assertion reflects the large number of studies in the literature on the precipitation and association properties of the caseins.There are, however, legitimate scientific goals in this kind of work other than the creation of artificial casein micelles, such as the elucidation of the mechanisms by which phosphoproteins profoundly influence the nucleation and growth of calcium phosphate phases. 

The most thorough study of the formation of artificial casein micelles is that of Schmidt and co-workers (1977 1979 Schmidt and Koops, 1977 Schmidt and Both, 1982 Schmidt and Poll, 1989), who not only studied the properties of the casein aggregates but also attempted to relate them to the solution conditions under which they were formed. In the precipitation of calcium phosphate from solution, the means by which solutions are mixed together is of crucial importance Schmidt et al. (1977) described a method in which four solutions were pumped simultaneously into a reaction vessel while keeping the pH constant. As a result of careful, slow mixing, the reproducibility of the size distributions of particles, measured by electron microscopy on freeze-fractured and freeze-etched specimens, was very good. In the first series of experiments, the objective was to produce milk like concentrations of the most important ions while... 

Casein micelles are remarkably stable structures. Milk may be boiled, sometimes for several hours, without coagulating the micelles. Also, the addition of CaCU to milk does not precipitate the micelles up to concentrations greatly in excess of that required to precipitate purified whole casein. On the other hand, micelles rapidly flocculate after treatment with chymosin, at or above room temperature, and casein... 

The effect of protein methylation (20% modification) on the stability of casein micelles was evaluated. The stability of the clJk micelle system towards Ca2+ precipitation was decreased when either or both proteins were methylated (see Figure 1). Methylation also resulted in a slight reduction in the stability of the /3-k casein micelle system (see Figure 2). 

The temperature-sensitive precipitation of unmodified and methylated /3-caseins in the presence of calcium was measured also (see Figure 3). Methylation caused an increase of up to 3°C in the precipitation temperature of calcium /3-caseinate. Results from rennet clotting of an asi-K casein micelle system indicated that replacing native asi-casein with the reductively methylated protein had little influence on clotting time, while replacing K-casein with its reductively methylated derivative re-... 

The caseins exist in milk as polydisperse aggregates ranging in size from ca. 40 to 220nm (3), but the size distribution of micelles depends upon the method of measurement. These casein micelles scatter light and are responsible for the whitish, opaque nature of skim milk. The casein micelles are also associated with a colloidal apatite comprised of calcium-phosphate-citrate (CPC) which has a stabilizing influence on the micelle structure. The colloidal CPC is in equilibrium with soluble CPC in the milk serum phase and is solubilized as the pH is reduced. Thus, as the pH is reduced to the isoelectric point of the caseins (4.6), the colloidal CPC solubilizes, and the caseins precipitate (143). This phenomenon should be kept in mind during some of the following discussions. 

Cryoprecipitation. When milk is frozen and stored at about — 10°C, the ionic strength of the liquid phase increases with a concomitant increase in [Ca " ] and a decrease in pH (to approximately 5.8) due to precipitation of calcium phosphates with the release of hydrogen ions (H ) (Chapter 5). These changes destabilize the casein micelles which precipitate when the milk is thawed. 

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