Comparison of bovine β-casein hydrolysis by PI and PIII-type proteinases from Lactobacillus lactis subsp. cremoris

The action of the cell-wall-associated proteinases from Lactococcus lactis subsp. cremoris strains H2 and SK112 on bovine ß-casein was compared. The proteinase from the H2 strain was characterised as a P_I-type proteinase since it did not hydrolyse a_s1-casein and the initial trifluoroacetic acid-soluble products of ß-casein hydrolysis were identical to those previously identified as hydrolysis products of P_I-type lactococcal proteinase action. The time-course of product formation by the proteinase from the H2 strain indicated that the bonds Tyr₁₉₃-Gln₁₉₄ and Gln₁₈₂-Arg₁₈₃ were the first to be hydrolysed. Cleavage of the bonds Gln₁₇₅-Lys₁₇₆, Ser₁₆₈-Lys₁₆₉, Ser₁₆₆-Gln₁₆₇ and Leu₁₆₃-Ser₁₆₄ was also very rapid. Four of the five bonds in ß-casein most susceptible to hydrolysis by the P_III-type proteinase from strain SK112 were different from those cleaved by the P_I-type proteinase, initial hydrolysis being at the sites Tyr₁₉₃-Gln₁₉₄, Leu₁₉₂-Tyr₁₉₃, Asp₄₃-Glu₄₄, Gln₄₆-Asp₄₇ and Phe₅₂-Ala₅₃. Early hydrolysis at the three sites in the N-terminal region of ß-casein, leading to cleavage of the N-terminal phosphopeptide and rapid precipitation of the residual fragment, represents a marked contrast to the action of P_I-type proteinases where cleavage at sites in the N-terminal region occurs only very slowly.