Clostridial collagenase (EC 220.127.116.11) catalyzes the hydrolysis of (Pro-Pro-Gly)5 at a minimum of three different rates, producing Pro-Pro, Gly-Pro-Pro and Gly-Pro-Pro-Gly, and various intermediate peptides. The intermediate and final products were separated by cation-exchange column chromatography and identified, and their rates of formation were measured. Pro-Pro was released most rapidly with formation of the tridecapeptide. After the initial release of the N-terminal Pro-Pro, hexa- and heptapeptides were formed in larger amounts than tri-, tetra-, nona- and decapeptides from the tridecapeptide. The rates of disappearance of the intermediates decreased in the order trideca- > deca- > and nona- > heptapeptide. The results indicate that the enzyme hydrolyzes inner linkages of the tridecapeptide having N- and C-terminal Gly residues, forming large peptides, preferentially to outer linkages, forming the tri- and tetrapeptides. © 1979.
Oshima, G., Shimabukubo, H., & Nagasawa, K. (1979). Mode of action of bacterial collagenase on a synthetic substrate, (Pro-Pro-Gly)5. BBA - Enzymology, 567(2), 392–400. https://doi.org/10.1016/0005-2744(79)90125-6