Hydrolysis of beta-casein and peptides by intracellular neutral protease of Streptococcus diacetylactis.

The endopeptidase activity of mesophilic streptococci was characterized further by investigating the specificity of an intracellular endopeptidase from Streptococcus diacetylactis for beta-casein, derived peptides, and bradykinin. The inhibitory action of phosphoramidon as well as direct determinations of metal content showed this enzyme was a metalloprotein. Hydrolysis of native beta-casein was relatively low. Peptides obtained from the fraction soluble at pH 4.6 led to the demonstration that Pro186-Ile187 and Ala189-Phe190 were hydrolyzed by the enzyme. Two peptides derived from beta-casein by the action of chymosin were hydrolyzed efficiently: we observed hydrolysis of Lys176-Ala177, Lys169-Val170, and Pro206-Ile207. The Pro7-Phe8 bond of bradykinin was hydrolyzed rapidly, showing that this enzyme was efficient for the hydrolysis of prolyl peptide bonds. The protease was slightly less sensitive to phosphoramidon than was thermolysin. Metal analyses showed the enzyme contained 580 microgram of zinc and 4,760 microgram of calcium per gram protein. This protease is thus a true metalloenzyme (E.C.3.4.24.4), and its action may complete the hydrolysis initiated by chymosin remaining active in cheese curd by hydrolyzing peptides released by chymosin.