High concentrations of sodium chloride facilitate the use of immobilized chymotrypsin for separating virgin forms of specific trypsin inhibitors

• Published in: Journal of Chromatography A 1999-08, Vol.852 (1), p.227-235
• Main Authors: Wilimowska-Pelc, Anna, Olichwier, Zofia, Kowalska, Jolanta, Gałuszka, Artur, Szuszkiewicz, Wiktor, Polanowski, Antoni, Wilusz, Tadeusz
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Aminoacids, peptides. Hormones. Neuropeptides; Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Chromatography, Affinity; Chromatography, High Pressure Liquid; Chromatography, Ion Exchange; Chymotrypsin - chemistry; Electrophoresis, Polyacrylamide Gel; Enzymes and enzyme inhibitors; Enzymes, Immobilized - chemistry; Fundamental and applied biological sciences. Psychology; Hydrolases; Molecular Sequence Data; Pancreas - enzymology; Proteins; Sequence Homology, Amino Acid; Sodium chloride; Sodium Chloride - chemistry; Swine; Trypsin inhibitors; Trypsin Inhibitors - chemistry; Trypsin Inhibitors - isolation & purification
• ISSN: 0021-9673
• DOI: 10.1016/S0021-9673(99)00441-0

It has been shown that specific trypsin inhibitors exhibit also antichymotrypsin activity in the presence of high NaCl concentrations. Taking advantage of this phenomenon a simple procedure of separation of the virgin forms of trypsin inhibitors from squash seeds and porcine pancreas (Kazal) was elaborated. In a typical experiment the inhibitor sample was loaded onto immobilized chymotrypsin equilibrated with 5 M NaCl at pH 8. After washing out unadsorbed material the virgin forms of inhibitors could be eluted either with water, buffer pH 8.0 or 0.02 M citrate buffer pH 2.6 containing no NaCl.