A signature protein-based method to distinguish Mediterranean water buffalo and foreign breed milk

A novel genetic variant at the α_s1-casein locus of water buffalo (WB), 8-residue shorter than its wild-type has been found and sequenced. The internal deletion of the peptide E³⁵KVNELsT⁴² was confirmed by the isolation of the junction peptide. The 8-residue deletion mutant has a molecular weight that is 919 Da less than that of the wild-type. The novel isoform with a unique f35-42 deletion could be the result of the skipping of exon 6, generating an exon 6-deleted variant of α_s1-casein. The wild-type and its shortened α_s1-casein forms were found to co-exist in many individual milk samples. In contrast, the 8-residue, internally deleted α_s1-casein variant did not occur in water buffaloes of the Mediterranean breed reared in Italy. Wild-type α_s1-casein has 6 to 8 phosphate groups (P) while the internally deleted form 6 and 7P per molecule.