A dipeptidyl aminopeptidase has been purified to apparent homogeneity from skin secretion of Xenopus laevis. This enzyme is a glycoprotein with a molecular mass of about 98 kDa. It hydrolyzes a variety of dipeptidyl-p-nitroanilides and oligopeptides containing proline, alanine or glycine as the second amino acid and is inhibited by diisopropylfluorophosphate. The pH optimum was found to be around 8, while at pH 6, substrates were cleaved at about one-third of the maximal rate. This dipeptidyl aminopeptidase has the specificity required for the cleavage of amino-terminal extensions preceding the sequence of caerulein and xenopsin in their respective precursors.