Purification and characterization of X-prolyl-dipeptidyl-aminopeptidase from Lactobacillus lactis and from Streptococcus thermophilus

Abstract

X-Prolyl-dipeptidyl-aminopeptidase recently was found in several lactic acid bacteria. This article describes the purification of the enzymes from Lactobacillus lactis and Streptococcus thermophilus and compares their characteristics. Enzymes from both strains are serine-peptidases. They both have a molecular weight of about 165,000 daltons, an isoelectric point near 4.5, and are constituted of two subunits. The pH optimum of the enzyme isolated from L. lactis is 7.0, whereas the enzyme from S. thermophilus possesses a broad pH optimum between 6.5 and 8.2 with glycyl-L-prolyl-aminomethylcoumarin as substrate. Below pH 5, both enzymes are unstable; however, that from S. thermophilus is more rapidly denatured. The enzyme from S. thermophilus is more sensitive to heat than the corresponding enzyme from L. lactis. Enzymes from the both strains have different specificities towards various substrates and are differently effected by metals, chelators, and other inhibitors. The importance of this enzyme for the metabolism of lactic acid bacteria is discussed.