The Isolation of Chymotrypsin-Like Enzymes by Affinity Chromatography Using Sepharose–4-Phenyibutyiamine

• Published in: Canadian Journal of Biochemistry
• Main Authors: Stevenson, Kenneth J., Landman, Amiram
• Format: Article in Journal/Newspaper
• Language: English
• Published: Canadian Science Publishing 1971
• Subjects: General Medicine; Alces alces
• Online Access: http://dx.doi.org/10.1139/o71-017; http://www.nrcresearchpress.com/doi/pdf/10.1139/o71-017

Affinity chromatography of chymotrypsin-like proteases on a column of Sepharose–4-phenylbutylamine (PBA) has been developed. Sepharose–PBA (Sepharose–NH∙[CH 2 ]4∙C 6 H 5 ) has been shown to selectively adsorb chymotrypsin α and B from weakly alkaline solutions and to allow to pass through unretarded porcine trypsin, bovine trypsinogen, and chymotrypsin α modified with active-site-directed irreversible inhibitors. Chymotrypsinogen A and bovine trypsin were only slightly retarded whereas a preparation of subtilisin was markedly retarded and separated into two distinct peaks. Sepharose–PBA has been utilized successfully for the selective isolation of chymotryps in-like proteases from extracts of moose pancreas (Alces alces).