Purification and characterization of ovine pancreatic elastase

Lýður Skúli Erlendsson, Hörður Filippusson*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Elastase was isolated from ovine pancreas and purified to homogeneity by two different procedures. One involved precipitation with ammonium sulphate, p-aminobenzamidine-Sepharose chromatography, CM-Sepharose ion exchange chromatography and S-300 Sephacryl chromatography. The other involved the direct adsorption of elastase by tri-l-alanyl-Sepharose chromatography and a CM-Sepharose step. The enzyme, which was produced in an inactive form in the pancreas, was activated with a trace of trypsin prior to chromatography. Ovine pancreatic elastase has an isoelectric point above pI 9.3 and its molecular mass is estimated at ~25 kDa. The optimal pH range for activity is between 8.0 and 8.4 and the enzyme is unstable at pH below 4.0 and above 10.0 and at temperatures above 65°C. The kinetic properties of the enzyme were determined with succinyl-Ala-Ala-Ala-p-nitroanilide as the substrate. K(m) and k(cat) K(m)-1 proved to be similar to the kinetic parameters of porcine elastase determined simultaneously. Copyright (C) 1998 Elsevier Science Inc.

Original languageEnglish
Pages (from-to)549-557
Number of pages9
JournalComparative Biochemistry and Physiology - B Biochemistry and Molecular Biology
Volume120
Issue number3
DOIs
Publication statusPublished - 1998

Other keywords

  • Affinity chromatography
  • Elastase
  • Ovine
  • Pancreas
  • Purification

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