Chemical and structural characterization of bacterially-derived casein peptides that impair milk clotting

Milk-clotting parameters are highly affected by hydrolysis of casein. Previously, it was shown that products of the hydrolysis of casein impair milk clotting, affecting both clotting time and curd firmness. One of these fractions is of particular interest since it is produced exclusively by enzymes of Streptococcus dysgalactiae. The present study aims to further investigate the chemical and structural properties of this fraction in an attempt to understand its influence on milk clotting. Preparations of this fraction, obtained from either S. dysgalactiae-infected glands or ex vivo inoculations with the same bacteria, were found to be identical. Mass spectrometry and Edman degradation analyses indicate that it comprises primarily β-CN83–209, generated by cleavage at a Val-Val peptide bond, presumably by bacterial thermolysin- or elastin-like proteases. A model offering a putative mechanism for interference with milk-clotting parameters through production of this fraction is presented.