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A simple method for the purification of dipeptidyl aminopeptidase IV from porcine kidney using HPLC
Author(s) -
Hutchinson D. W.,
Woolfitt A. R.
Publication year - 1986
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.1130010403
Subject(s) - chemistry , chromatography , aminopeptidase , high performance liquid chromatography , simple (philosophy) , biochemistry , amino acid , philosophy , epistemology , leucine
Depeptidyl 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.