Colagenasa from Clostridium histolyticum

La colagenasa es activada por cuatro átomo-gramos de calcio por mol de enzima. Es inhibida por el ácido etilenglicol-bis(beta-aminoetil éter) - N, N, N′,N′-tetraacético, el beta-mercaptoetanol, el glutatión, el ácido tioglicólico y la 8-hidroxiquinolina.

Crude collagenases are mixtures of enzymes (mostly proteases) secreted by C. histolyticum. Effective release of cells from tissue requires the action of both 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.

Una unidad de digestión de colágeno (CDU) libera péptidos del colágeno del tendón de Aquiles bovino que equivalen en la reacción de color de la ninhidrina a 1,0 μmoles de leucina en 5 horas a pH 7,4 y 37 °C en presencia de iones calcio. Una unidad de hidrólisis FALGPA hidroliza 1,0 μmoles de furilacriloil-Leu-Gly-Pro-Ala por minuto a 25°C. Una unidad de proteasa neutra hidroliza la caseína para producir color equivalente a 1,0 μmoles de tirosina por 5 horas a pH 7,5 y 37°C. Una unidad de clostripaína hidroliza 1,0 μmoles de BAEE por minuto a pH 7,6 y 25°C en presencia de DTT.

Collagenase from Clostridium histolyticum has been used:

• for the recovery of neutrophils from collagen gels
• for the dissociation of breast tumor biopsies into single cells
• for the isolation of primary human endometrial stromal cells(HESC) from endometrial biopsies

Collagenase from Clostridium histolyticum may be used for the isolation of cells from many types of animal tissue. It may be used for the disaggregation of human tumor, mouse kidney, human adult/fetal brain, lung and many other tissues. It may also be used in liver and kidney perfusion studies, for the digestion of pancreas, isolation of nonparenchymal rat liver cells and for hepatocyte preparation.

Collagenase from Clostridium histolyticum comprises of collagenases G and H. Collagenase has N-terminal activator domain and C-terminal peptidase domain. The collagen binding domains (CBD) and polycystic kidney disease-like (PKD-like) domain resides in the peptidase domain. The collagenase module undergoes structural changes upon calcium binding.

Product sold on the basis of collagen digestion units (CDU).

Collagenase may be dissolved in TESCA buffer (50 mM TES, 0.36 mM Calcium chloride, pH 7.4 at 37 °C) at 0.05-0.1 mg/mL concentration.