High concentrations of sodium chloride facilitate the use of immobilized chymotrypsin for separating virgin forms of specific trypsin inhibitors.
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.
Wilimowska-Pelc
A
A
Institute of Biochemistry and Molecular Biology, University of Wrocław, Poland.
Olichwier
Z
Z
Kowalska
J
J
Gałuszka
A
A
Szuszkiewicz
W
W
Polanowski
A
A
Wilusz
T
T
D000595:Amino Acid Sequence; D000818:Animals; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D002918:Chymotrypsin / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D004800:Enzymes, Immobilized / Q000737:chemistry*; D008969:Molecular Sequence Data; D010179:Pancreas / Q000201:enzymology; D017386:Sequence Homology, Amino Acid; D012965:Sodium Chloride / Q000737:chemistry*; D013552:Swine; D014361:Trypsin Inhibitors / Q000737:chemistry / Q000302:isolation & purification*
D004800:Enzymes, Immobilized; D014361:Trypsin Inhibitors; D012965:Sodium Chloride; D002918:Chymotrypsin