A simple method for the purification of dipeptidyl aminopeptidase IV from porcine kidney using HPLC.

Dipeptidyl aminopeptidase IV (E.C. 3.4.14.5) has been purified to homogeneity using "soft-column" chromatography on Sephadex G-200, DEAE-cellulose, gly-pro-AH-Sepharose and finally HPLC on TSK G-3000SW. The purification by HPLC is fast and gives a better yield of pure enzyme than procedures which have been described previously. The enzyme obtained in this manner has a high specific activity (86 U/mg using a saturating level of gly-pro-4-nitroanilide as substrate) and was free of contaminating peptidase activities. Our preparation of DAP IV is suitable for peptide sequencing studies by fast atom bombardment mass spectrometry.