Purification and some properties of camel carboxypeptidase B

Mol Cell Biochem. 1999 Nov;201(1-2):105-10. doi: 10.1023/a:1007012518250.

Abstract

A carboxypeptidase B-like enzyme was purified 116-fold with a recovery of activity of 29% from a crude extract of camel pancreas by a four-step procedure consisting of two anion exchange chromatographies in succession, gel filtration and hydrophobic interaction chromatography. The enzyme was homogeneous on SDS and non-denaturing gel electrophoresis and on gel isoelectric focusing. Its molecular mass was found to be 31.5 kDa and its isoelectric point was estimated as 6.1. It was active towards a number of substrates that are cleaved by carboxypeptidases B from other species and was also susceptible to inhibition by inhibitors of such enzymes. The camel enzyme showed a pH optimum of 8.0 and it was seen to be a relatively potent kininase in vitro. The enzyme purified in this study was very similar to carboxypeptidases B isolated from other species in size, charge, substrate specificity and susceptibility to inhibition and thus it can be identified as camel carboxypeptidase B.

MeSH terms

  • Animals
  • Camelus
  • Carboxypeptidase B
  • Carboxypeptidases / antagonists & inhibitors
  • Carboxypeptidases / isolation & purification*
  • Carboxypeptidases / metabolism*
  • Chromatography, Ion Exchange / methods
  • Edetic Acid / pharmacology
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Inhibitors / pharmacology
  • Enzyme Stability
  • Hydrogen-Ion Concentration
  • Hydrolysis
  • Pancreas / enzymology
  • Substrate Specificity

Substances

  • Enzyme Inhibitors
  • Edetic Acid
  • Carboxypeptidases
  • Carboxypeptidase B