P. Lechevalier et al., PURIFICATION AND PARTIAL CHARACTERIZATION OF CHYMOTRYPSIN-LIKE PROTEASES FROM THE DIGESTIVE GLAND OF THE SCALLOP PECTEN-MAXIMUS, Comparative biochemistry and physiology. B. Comparative biochemistry, 110(4), 1995, pp. 777-784
Three variants of a chymotrypsin-like protease were purified from scal
lop digestive glands successively by ion-exchange, gel filtration and
high-performance liquid chromatographies. Enzyme activity was detected
using succinyl-Ala-Ala-Pro-Phe-p-nitroanilide as a specific synthetic
substrate for chymotrypsin, This proteinase was inhibited by chymosta
tin, diisopropylfluorophosphate and phenylmethylsulfonyl fluoride, Est
imated molecular mass of the purified enzyme is around 32 kDa. These i
soenzymes exhibit very low activities in hydrolyzing small synthetic s
pecific substrates used for trypsic, elastolytic and collagenolytic me
asurements and referred mainly to a chymotrypsin-like proteinase, Very
few differences were measured concerning pH profiles among the three
isoenzymes. Stability is higher at low temperature for two variants. A
n N-terminal analysis was performed on one variant (B) among the three
isoenzymes. The alignment of the N-terminal amino acid sequence indic
ates some homologies with abalone chymotrypsin-like protein and arthro
pod chymotrypsin proteases as well as with vertebrate serine protease
counterparts (trypsin, chymotrypsin and elastase).