Affinity chromatography of trypsin and related enzymes. III. Purification of Streptomyces griseus trypsin using an affinity adsorbent containing a tryptic digest of protamine as a ligand.
A new, simple method has been developed for the purification of Streptomyces griseus trypsin [EC 3.4.21.4] from Pronase. Only a single operation of affinity chromatography on an agarose derivative, which was easily prepared by coupling a tryptic digest of salmine to cyanogen bromide-activated Sepharose 4B, was required. A high degree of homogeneity was demonstrated for the purified enzyme by disc electrophoresis, SDS-polyacrylamide gel electrophoresis and gel filtration, as well as by active-site titration. The behavior of a carboxypeptides B [EC 3.4.12.3]-like enzyme present in Pronase is also discussed.
D001665:Binding Sites; D002268:Carboxypeptidases / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D011402:Pronase / Q000302:isolation & purification*; D011479:Protamines; D011485:Protein Binding; D013305:Streptomyces griseus / Q000201:enzymology*; D014357:Trypsin / Q000302:isolation & purification* / Q000378:metabolism
D011479:Protamines; D002268:Carboxypeptidases; D014357:Trypsin; D011402:Pronase