Preparation of β-trypsin by affinity chromatography of enterokinase-activated bovine trypsinogen

✍ Scribed by Juris J. Liepnieks; Albert Light

• Book ID: 102982105
• Publisher: Elsevier Science
• Year: 1974
• Tongue: English
• Weight: 609 KB
• Volume: 60
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(74)90247-4

✦ Synopsis

The preparation of /3-trypsin can be simply accomplished from the activation of bovine trypsinogen with partially purified enterokinase in the presence of STI-Sepharose. Enterokinase catalyzes the specific cleavage of lysine 6-isoleucine 7 and the presence of ST1 prevents autolysis of /3-trypsin by forming a stable inactive complex. The ST1 immobilized to Sepharose is suitable for the subsequent purification of the activation mixture by affinity chromatography. Inert protein and contaminants are removed with a buffer at pH 4.5. A change to a buffer at pH 2.6 or the introduction of a pH gradient leads to the recovery of highly purified P-trypsin. Thr procedure produces /3-trypsin in a 70-75s yield, which is essentially a theoretical recovery, and all operations can be completed within 6 hr.