Purification and partial characterization of camel anionic chymotrypsin.

An anionic chymotrypsin-like enzyme was isolated from a crude extract of camel pancreas by a three-step procedure consisting of anion-exchange chromatography, gel filtration, and hydrophobic interaction chromatography. The purified enzyme was homogeneous on native and SDS gel electrophoresis and on gel isoelectric focusing. Its molecular mass was estimated as 28.5 kDa and its isoelectric point was found to be 4.4. The enzyme differed markedly from bovine chymotrypsin A in its substrate specificity, showing considerably lower values of the specificity constant for its action on tyrosine, tryptophan, and phenylalanine esters. Its pH optimum was found to be 7.8. It showed lower kininase activity and was more susceptible to inhibition by a number of inhibitors than the bovine cationic chymotrypsin. On the other hand, the camel enzyme showed a much greater hydrolytic activity than the bovine enzyme toward a leucine ester. In terms of its size, charge, and substrate specificity the camel enzyme was very similar to anionic chymotrypsins that have been isolated from other species and thus appears to be a camel anionic chymotrypsin.