SOURCE. CI Histolyticum.
SPECIFICATIONS.
Chromatographically purified, salt-free, lyophilized water soluble powder.
Store at 5° C. Will remain stable 9 to 12 months at 5° C.
1000-2000 units per mg of protein. One unit will liberate a ninhydrin color equivalent to 1 µM of L-leucine at pH 7.5 in 5 hours at 37°C. Bovine achilles tendon collagen (EPC No. C204) is the substrate.
Also, 8-12 FALGPA units per mg of protein where 1 unit will hydrolyze 1 µmole of furanacryloyl-Leu-Gly-Pro-Ala at pH 7.5 per minute. (53).
Clostripain; <0.1 units per mg of protein
Tryptic Activity: <0.3 units per mg of protein.
Protease Activity: <1 units per mg of Protein.
PREPARATION.
Prepared by our modifications of the Keller and Mandl (20). In the method, crude collagenase is chromatographed sequentially on SP-Sephadex* at pH 6.0 and on DEAE-Cellulose at pH 8.0 These ion exchange steps remove most of the non-specific protein. The high collagenolytic fraction (low protease activity) is further purified by gel diffusion on Sephadex-G200. Non-specific protease is removed by affinity chromatography. Salt-free, lyophilized.