Immobilized anhydrochymotrypsin as a biospecific affinity adsorbent for the peptides produced by chymotryptic hydrolysis.
Anhydrochymotrypsin immobilized on Sepharose specifically adsorbed various peptides containing L-tryptophan, L-tyrosine, or L-phenylalanine residues at their carboxy-termini. These peptides correspond to the specific products of chymotryptic cleavage of polypeptides. A mixture of the chymotryptic peptides once adsorbed on the column could be effectively separated by eluting them with a pH gradient. This adsorbent, on the other hand, showed no substantial affinity toward the substrate-type peptides that contain the aromatic amino acid(s) within the peptide chain, or toward the C-terminal leucine peptides. By taking advantage of this unique property of anhydrochymotrypsin-Sepharose in combination with reversed-phase high-performance liquid chromatography, we have succeeded in separating the C-terminal peptides from chymotryptic digests of reduced and S-carboxymethylated bovine insulin and from tryptic digests from reduced and S-carboxymethylated Streptomyces subtilisin inhibitor.
Kumazaki
T
T
Department of Biochemistry, Faculty of Pharmaceutical Sciences, Hokkaido University.
Fujitani
A
A
Terasawa
K
K
Shimura
K
K
Kasai
K
K
Ishii
S
S
D000327:Adsorption; D000596:Amino Acids / Q000032:analysis; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D002918:Chymotrypsin; D004800:Enzymes, Immobilized; D006868:Hydrolysis; D010455:Peptides / Q000032:analysis*
D000596:Amino Acids; D004800:Enzymes, Immobilized; D010455:Peptides; D002918:Chymotrypsin; C000653:anhydrochymotrypsin