Description

• General description: Clostridium histolyticum is a pathogenic clostridium that produces crude collagenases. This is a mixture of enzymes containing collagenase, non-specific proteases and clostripain.[1]
• Application: Collagenase may be used:[2][1][3][4][5][6]for the preparation of arterial tissue for the study of advanced glycosylation end products (AGE)for use along with other proteases for the disaggregation of human tumor, mouse kidney, human brain, lung epithelium and many other tissues.in liver and kidney perfusion studies, digestion of pancreas, and isolation of nonparenchymal hepatocytes.for the preparation of viable hepatocytes from rat liver and for the isolation of fat cells from rat adipose tissue
• Biochem/physiol Actions: Collagenase is activated by four gram atom calcium per mole enzyme. It is inhibited by ethylene glycol-bis(β-aminoethyl ether) - N, N, N′,N′-tetraacetic acid, β-mercaptoethanol, glutathione, thioglycolic acid and 8-hydroxyquinoline. Collagenase enzymes and neutral protease plays an important role in the effective release of cells from tissue.[7] Collagenase recognizes the sequence -R-Pro-8-X-Gly-Pro-R-, where X represents a neutral amino acid. [8]
• Unit Definition: One collagen digestion unit (CDU) liberates peptides from collagen from bovine achilles tendon equivalent in ninhydrin color to 1.0 μmole of leucine in 5 hours at pH 7.4 at 37 °C in the presence of calcium ions. One FALGPA hydrolysis unit hydrolyzes 1.0 μmole of furylacryloyl-Leu-Gly-Pro-Ala per min at 25°C. One Neutral Protease unit hydrolyzes casein to produce color equivalent to 1.0 μmole of tyrosine per 5 hr at pH 7.5 at 37°C. One Clostripain Unit hydrolyzes 1.0 μmole of BAEE per min at pH 7.6 at 25°C in the presence of DTT.
• Analysis Note: Also contains clostripain, nonspecific neutral protease and tryptic activities.