The Isolation of Trypsin and Elastase from Moose Pancreas ( Alces alces ) by Affinity Chromatography on Lima-Bean Protease Inhibitor – Sepharose Resin

• Published in: Canadian Journal of Biochemistry
• Main Authors: Lievaart, Paul A., Stevenson, Kenneth J.
• Format: Article in Journal/Newspaper
• Language: English
• Published: Canadian Science Publishing 1974
• Subjects: General Medicine; Alces alces
• Online Access: http://dx.doi.org/10.1139/o74-091; http://www.nrcresearchpress.com/doi/pdf/10.1139/o74-091

Chymotrypsin, trypsin, and elastase were isolated from pancreas glands of the moose (Alces alces) by acid extraction, (NH 4 ) 2 SO 4 fractionation, and affinity chromatography conducted sequentially on 4-phenylbutylamine (PBA) – Sepharose and lima-bean protease inhibitor (LBI) – Sepharose. The removal of chymotrypsin on PBA–Sepharose led to the coabsorption of trypsin and elastase on to LBI–Sepharose. The binding site of elastase was identified as the 'chymotrypsin-site' of LBI. Elastase and trypsin were each selectively desorbed from LBI–Sepharose. These proteases exhibited identical elution profiles during carboxymethylcellulose chromatography and had similar migration rates on polyacrylamide disc gel electrophoresis. Moose elastase contained some trypsin activity whereas the trypsin preparation was pure based on comparison of ionograms of peptides derived from the proteolytic digestion of glucagon by moose chymotrypsin, trypsin, elastase, bovine trypsin, and porcine elastase.