Abstract
Three new protease inhibitors were isolated and purified about 200-fold from hemolymph of silkworm larvae, Bombyx mori, using ion-exchange and affinity chromatography. Two of the three inhibitors were basic proteins (SCI-I had pI 9.4 and SCI-11 had pI 9.6) and one was acidic (SCI-TII had pI 4.0). The molecular weight of each inhibitor was determined to be 7, 000 by the sedimentation equilibrium method. The amino acid compositions of the inhibitors were similar except for the contents of Asp, Glu, Ile, Len, and Lys. Val, His, and Trp were not present in the inhibitors and Met appeared only in SCI-III. The CD spectra of the inhibitors were all similar and indicated a low content of α-helical structure (10% at most). Each inhibitor could inhibit the protease and esterase activities of bovine α-chymotrypsin at a one-to-one molar ratio, and the dissociation constants were 3.1×10-9 M for SCI-I and II and 1.3×10-8 M for SCI-III. Only SCI-II showed a weak inhibitory activity against bovine trypsin. Subtilisin BPN' and papain were not inhibited by these inhibitors.
