Collagenase II (Synonyms: Collagenase; collagen hydrolase)

Collagenase Ⅱ is a matrix metalloproteinase (MMPs) used for tissue dissociation and the preparation of liver, bone, thyroid, heart, and salivary gland cells^[1,2]. Collagenase can cleave the bond between neutral amino acids and glycine in the Pro-X-Glyc-Pro sequence, which is common in collagen ^[3]. The enzyme activity of collagenase requires the presence of divalent cations such as Ca²⁺ and Zn²⁺ for activation^[4]. Collagenase activity is inhibited by agents such as ethylenediaminetetraacetic acid (EDTA), dithiothreitol (DTT), detergents, hexachlorocyclohexane, and heavy metal ions (Hg²⁺, Pb²⁺, Cd²⁺) ^[5]. Collagenase is relatively mild and exhibits good dissociation ability at physiological temperature and pH without the need for mechanical stirring or special equipment. The optimal pH for Collagenase II is 6.3-8.5, and its enzymatic activity is ≥125 CDU/mg solid (CDU = Collagen Digestion Units).

There are approximately five common types of collagenase ^[6]:

(1) Collagenase Type I (GC19589): Contains relatively uniform multiple enzymatic activities (including collagenase, caseinase, clostridiopeptidase, and trypsin activities), and is commonly used for the dissociation of epithelial, lung, adipose, and adrenal tissue cells.

(2) Collagenase Type II (GC19588): Contains higher clostridiopeptidase activity and is typically used for the dissociation of liver, bone, thyroid, heart, and salivary gland tissue cells.

(3) Collagenase Type III (GC19590): Contains lower protease activity and is commonly used for the dissociation of mammary gland cells.

(4) Collagenase Type IV (GC19591): Contains lower trypsin activity and is typically used for the preparation of islet cells or experiments that require the preservation of cell surface receptors.

(5) Collagenase Type V (GC19592): Partially purified, containing higher collagenase and caseinase activities, and is typically used for the dissociation of pancreatic islet tissue and connective tissue dissociation.

References:
[1] Sekhon B S. Matrix metalloproteinases–an overview[J]. Research and Reports in Biology, 2010: 1-20.
[2] Alipour H, Raz A, Zakeri S, et al. Therapeutic applications of collagenase (metalloproteases): A review[J]. Asian Pacific Journal of Tropical Biomedicine, 2016, 6(11): 975-981.
[3] Trabelsi O, Dumas V, Breysse E, et al. In vitro histomechanical effects of enzymatic degradation in carotid arteries during inflation tests with pulsatile loading[J]. Journal of the mechanical behavior of biomedical materials, 2020, 103: 103550.
[4] Eming S, Smola H, Hartmann B, et al. The inhibition of matrix metalloproteinase activity in chronic wounds by a polyacrylate superabsorber[J]. Biomaterials, 2008, 29(19): 2932-2940.
[5] Daboor S M, Budge S M, Ghaly A E, et al. Extraction and purification of collagenase enzymes: a critical review[J]. Am. J. Biochem. Biotechnol, 2010, 6(4): 239-263.
[6] Alipour H, Raz A, Zakeri S, et al. Therapeutic applications of collagenase (metalloproteases): A review[J]. Asian Pacific Journal of Tropical Biomedicine, 2016, 6(11): 975-981.