Collagénase from Clostridium histolyticum

The enzyme has been used in the isolation of mast cells from adult mice. This study generated a large number of connective tissue-type mast cells by the culture of murine fetal skin cells. The product has also been used in the in vitro biodegradation study of Bombyx mori silk fibroin fibers and films, by exposing them to the enzyme for varied time periods.

Collagenase from Clostridium histolyticum, or Clostridiopeptidase A, has been used in a study to assess contact dermatitis with clostridiopeptidase A contained in Noruxol ointment. Clostridiopeptidase A has also been used in a study to investigate the early and successful enzymatıc debridement via collagenase application to pinna in a preterm neonate.

Collagenase from Clostridium histolyticum has been used in determining the degradation rate of collagen biomaterials. It may also be used to determine the enzymatic degradation and enzymatic resistance by the liberated L-leucine measurements.

La collagénase est activée à l′aide de quatre atome-grammes de calcium par mole d′enzyme. Elle est inhibée par l′acide éthylèneglycol-bis(bêta-aminoéthyléther)-N,N,N′,N′-tétraacétique, le bêta-mercaptoéthanol, le glutathion, l′acide thioglycolique et la 8-hydroxyquinoline.

Effective release of cells from tissue requires the action of collagenase enzymes and the neutral protease. Collagenase is activated by four gram atom calcium (Ca²⁺) per mole enzyme. The culture filtrate is thought to contain at least 7 different proteases ranging in molecular weight from 68-130 kDa. The pH optimum is 6.3-8.8. The enzyme is typically used to digest the connective components in tissue samples to liberate individual cells. Ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid (EGTA)4; β-mercaptoethanol; glutathione, reduced; thioglycolic acid, sodium; and 2,2′-dipyridyl; 8-hydroxyquinoline are known to inhibit the enzyme activity.

Une unité de digestion de collagène (UDC) libère une quantité de peptides à partir du collagène d′un tendon d′Achille de bovin équivalant, en termes de couleur avec la méthode à la ninhydrine, à 1,0 μmole de leucine en 5 heures à pH 7,4 et à 37 °C en présence d′ions calcium. Une unité d′hydrolyse de FALGPA hydrolyse 1,0 μmole de furylacryloyl-Leu-Gly-Pro-Ala par minute à 25 °C. Une unité de protéase neutre hydrolyse la caséine et produit une couleur équivalente à 1,0 μmole de tyrosine en 5 heures à pH 7,5 et à 37 °C. Une unité de clostripaïne hydrolyse 1,0 μmole de BAEE par minute à pH 7,6 et à 25 °C en présence de DTT.