Phosphopeptides from Grana Padano cheese: nature, origin and changes during ripening.

Casein phosphopeptides (CPP) which develop in Grana Padano cheese at different ages were isolated by precipitation with Ba2+ and analysed by HPLC. Profiles were complex throughout the period between 4 and 38 months. CPP in a cheese sample 14 months old were identified by a combination of fast atom bombardment-mass spectrometry and Edman degradation.

Evaluation of the stable reaction products of histidine with formaldehyde or with other carbonyl compounds in dairy products.

Combined formaldehyde (FA) can be evaluated in cheese and other dairy products by determining spinacine (6-carboxy-1,2,3,4-tetrahydroimidazopyridine), an imino acid arising from the reaction of FA with the alpha-amino group of histidine. Other carbonyl compounds usually produced by fermentations can react themselves with histidine, forming molecules which interfere with the determination of spinacine. A sensitive (minimum detectable amount, 5 fmol spinacine) and interference-free HPLC method, with precolumn derivatization with o-phtalaldehyde (OPA) and 9-fluorenylmethylchloroformate (FMOC), to evaluate these histidine reaction-compounds in dairy products is described.

Gel electrophoresis and immunoblotting for the detection of casein proteolysis in cheese.

The whole N fraction of six samples of hard and semi-hard pressed cheeses was analysed using PAGE, polyacrylamide gel isoelectric focusing and immunoblotting with polyclonal antibodies against beta- and alpha s1-casein. The origin of some electrophoretic bands corresponding to peptides produced from the enzymic degradation of the casein fractions was established. A number of these peptides were also present in the in vitro hydrolysates of casein with plasmin and chymosin.

Identification of spinacine as the principal reaction product of gamma-casein with formaldehyde in cheese.

Formaldehyde is added to milk in the production of grana cheese as an antimicrobial agent. In order to study the fate of the formaldehyde, a grana cheese preparation was made using 14C-labelled formaldehyde. The 14C-activity in the cheese was found to be mainly associated with the caseins, but it was not uniformly distributed among the different fractions (alpha s, beta- and gamma-caseins).