Purification of carboxypeptidase B by metal chelation affinity chromatography.
Norwegian lobster carboxypeptidase B (CPB) was purified in one step using immobilized metal chelate affinity chromatography (IMAC). The separation is based on the property that CPB has a high affinity for metal ions such as Cu2+. The CPB was purified from an hepatopancreas extract containing several endo- and exo-proteolytic activities. Its homogeneity was demonstrated by SDS-electrophoresis and isoelectric focusing in immobilized pH gradients. The implication of hydrophobic interaction between this enzyme and the IDA-Cu2+ gel is postulated.
Márquez-Méndez
M
M
Laboratoire de Technologie des Séparations Université de Technologie de Compiègne, France.
D000818:Animals; D043424:Carboxypeptidase B; D002268:Carboxypeptidases / Q000737:chemistry / Q000302:isolation & purification*; D002614:Chelating Agents; D002846:Chromatography, Affinity / Q000379:methods; D003300:Copper; D007525:Isoelectric Focusing; D008099:Liver / Q000201:enzymology; D008970:Molecular Weight; D008121:Nephropidae; D010179:Pancreas / Q000201:enzymology; D013379:Substrate Specificity
D002614:Chelating Agents; D003300:Copper; D002268:Carboxypeptidases; D043424:Carboxypeptidase B