PURIFICATION AND CHARACTERIZATION OF A POSTPROLINE DIPEPTIDYL AMINOPEPTIDASE FROM STREPTOCOCCUS-CREMORIS AM2

The present study describes the purification of a post-proline dipeptidyl aminopeptidase from the cytoplasm of Streptococcus cremoris AM2. On the basis of its elution from a calibrated Sephadex G200 column, the enzyme had a molecular weight of 117000 and exhibited a broad pH optimum activity between 6.0 and 9.0. The activity was most comprehensively inhibited by phenylmethylsulphonylfluoride and more modestly inhibited by 1, 10-phenanthroline and 8-hydroxyquinoline but not by EDTA. A range of peptides containing either proline or alanine as the penultimate amino acid residue could act as substrates. The presence of proline on the carboxy side of the scissile bond prevented hydrolysis. However the enzyme could release Pro-Pro from Pro-Pro-Gly-Phe-Ser-Pro. The significance of this substrate specificity is considered in the context of removal of either single proline residues or prolyl-proline sequences from oligopeptides during cheese ripening. © 1990, Proprietors of Journal of Dairy Research. All rights reserved.