Purification and characterization of chymotrypsin, trypsin and elastase like proteinases from cod (Gadus morhua L.).
1. Chymotrypsin, trypsin and elastase have been purified from the pyloric caeca of cod. 2. The enzymes were separated by affinity/hydrophobic chromatography on phenyl-butyl-amine (PBA) substituted sepharose. Chymotrypsin eluted in two separate isozyme fractions whereas trypsin and elastase eluted in separate fractions consisting of two closely-related polypeptide chains as revealed by SDS-polyacrylamide electrophoresis and isoelectric focusing. 3. The cod enzymes consist of single polypeptide chains with apparent molecular weights of about 27,000 Da as shown by denaturing polyacrylamide gel electrophoresis. 4. The cod proteinases were retarded on gel filtration media. The retardation increased with increasing pressure. 5. Isoelectric focusing analysis shows that the cod enzymes have isoelectric points in the range between 5 and 7. 6. The cod proteinases are rapidly inactivated when stored at low pH's.
Department of Biochemistry, University of Bergen, Norway.
A J
AJ
Raae
B T
BT
Walther
D000818:Animals; D002432:Cecum / Q000201:enzymology; D002918:Chymotrypsin / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D005399:Fishes / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008970:Molecular Weight; D010196:Pancreatic Elastase / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D014357:Trypsin / Q000302:isolation & purification* / Q000378:metabolism; D014361:Trypsin Inhibitors
D014361:Trypsin Inhibitors; D002918:Chymotrypsin; D010196:Pancreatic Elastase; D014357:Trypsin