Pancreatic proteolytic enzymes from carp (Cyprinus carpio)-I purification and physical properties of trypsin, chymotrypsin, elastase and carboxypeptidase B

1. 1. Trypsin, chymotrypsin, elastase and carboxypeptidase B were purified from carp pancreas by ion-exchange and affinity chromatography. All zymogens that existed in the initial extract of the pancreas were converted to the respective enzymes by self-activation prior to purification. 2. 2. The purified trypsin, chymotrypsin and elastase consisted respectively, of four, three and two electrophoretically distinct but enzymatically active fractions Carboxypeptidase B consisted of one major and one minor band. 3. 3. The approximate molecular weights of trypsin, chymotrypsin and elastase were 25,000 and carboxypeptidase B 34,000. The enzymes are not stable at a low pH but retain their activity at a neutral pH in the presence of Ca²⁺. 4. 4. Amino acid analysis of trypsin, elastase and chymotrypsin revealed high similarities to the respective bovine or porcine enzymes. No such similarity was detected in carboxypeptidase B. 5. 5. Amino terminal sequence analysis of the purified chymotrypsin resulted in one main chain resembling that of the B-chain of bovine chymotrypsin. However, the existence of an additional minor chain cannot be excluded.