Proteomic characterization of kefir milk by two-dimensional electrophoresis followed by mass spectrometry.

Kefir is a type of fermented milk obtained thanks to the introduction of "kefir grains" in mammalian milk. Kefir grains consist of lactic and acetic acid bacteria and yeasts in alternative proportions that are held together by a matrix of complex sugars known as "kefiran." Thanks to the fermentative process, the kefir milk is rich in nutraceutical substances such as amino acids, vitamins, and mineral salts. The most valuable compounds of kefir fermentation are mainly lactic acid, exopolysaccharides, and bioactive peptides, the resulting products of proteolytic release from milk proteins (caseins and whey proteins). Among the nutraceutical properties of kefir are antimicrobial and antitumor activity, immunomodulating effect, and cholesterol-lowering effect. Therefore, in light of these intriguing properties of kefir milk, in this work, a proteomic analysis, by two-dimensional electrophoresis followed by mass spectrometry, has been performed. As a result, milk-derived polypeptides were identified in commercial kefir milk from organic farming. In particular, polypeptides deriving from κ-, αs1 -, and αs2 -caseins that may have potentially beneficial effects on human health have been detected.

A comparison among ß-caseins purified from milk of different species: Self-assembling behaviour and immunogenicity potential.

Caseins are a family of proteins constituted by α-caseins (αs-1 and αs-2 caseins), ß-caseins and κ-caseins. ß-caseins, in particular, show a temperature and concentration-dependent self-assembling behaviour. Recently, ß-casein micelles have been proposed as natural nanocarriers for the delivery of hydrophobic compounds, promoting their bioavailability. Until now, all studies regarding both chemical-physical characterization and applications of ß-caseins have employed the protein of bovine origin. However, it could be interesting to exploit the use of ß-caseins from other milk sources for their potential encapsulation ability and immunogenicity but, at present, no information on the self-assembling behaviour is available for ß-caseins from the milk of species different from bovine. In this work, for the first time, ß-caseins from human milk and from donkey, goat, and sheep milk were purified and their self-assembling behaviour was compared to that of a commercial bovine ß-casein, the only one for which the concentration and temperature aggregation behaviour is known. Furthermore, a preliminary evaluation of the immunogenicity potential of ß-casein from other milk sources has been performed by cross-reaction experiments using anti-ß-casein antibodies from bovine origin. The results indicated a similar self-assembling profile among all ß-caseins examined compared to the bovine ß-casein, suggesting the possible use of ß-casein from other milk sources as nanocarriers. Since donkey and human ß-casein do not cross-react with bovine anti-ß-casein antibodies, they could be particularly interesting for the development of self-assembling systems with lower hypoallergenic potential.