Collagenase I (Synonyms: Collagenase; collagen hydrolase)

The collagenase currently has approximately four types: Type I collagenase, Type II collagenase, Type III collagenase, and Type IV collagenase, which have specific applications: Type I Collagenase: Contains a relatively uniform variety of enzyme activities (including collagenase, caseinase, clostripain, and trypsin activity). It is commonly used for the preparation of epithelial cells and tissue cells. Type II Collagenase: Contains higher clostripain activity and is typically used for the preparation of cells from tissues such as heart, bone, muscle, thymus, and cartilage. Type III Collagenase: Contains lower protease activity and is frequently used for the preparation of mammary gland cells. Type IV Collagenase: Contains low trypsin activity and is usually employed for the preparation of pancreatic islet cells or for cell preparation experiments where receptor integrity needs to be maintained.

Collagenase I is classified as a proteinase derived from Bacillus histolyticus, having the capability to cleave the bonds between neutral amino acid and glycine residues within Pro-X-Gly-Pro sequences, which are prominently found in collagen proteins.

Collagenase I is relatively mild, with an initial balance of collagenase, caseinase, clostridium protease, and trypsin activity, and is well dissociated at physiological temperature and pH. The optimal pH of collagenase I is 7 ~ 8.

It has the unique ability to digest natural collagen and denatured collagen. Unique among proteases, collagenase I possesses the ability to break down the triple-helical natural collagen fibers that are abundant in connective tissues like skin, tendons, blood vessels, and bones. The proteolytic degradation facilitated by collagenase is employed in the enzymatic digestion of human tumors, mouse kidneys, adult and fetal brains, and various additional tissues, encompassing those featuring epithelial constituents. It acts exclusively on procollagen, breaking it and then being hydrolyzed by other proteases, without hydrolyzing fibrin and globulin. Detergents, hexachlorocyclohexane and heavy metal ions can reduce enzyme activity[1-5]. Collagenase I (Intravenous injection) could transiently reduceinterstitial fluid pressure (IFP) by digestion of collagen in tumors and increase tumor accumulation and gene expression of lipoplex[6].

Activity definition: One FALGPA hydrolysis unit hydrolyzes 1.0 µmole of furylacryloyl-Leu-Gly-Pro-Ala per min at pH 7.5 at 25 °C in the presence of calcium ions. One neutral protease unit hydrolyzes casein to produce color equivalent to 1.0 µmole tyrosine per 5 hr at pH 7.5 at 37℃. One clostripain unit hydrolyzes 1.0 µmole of BAEE per min at pH 7.6 at 25℃ in the presence of DTT.

References:
[1]. Pilcher BK, Sudbeck BD, et,al. Collagenase-1 and collagen in epidermal repair. Arch Dermatol Res. 1998 Jul;290 Suppl:S37-46. doi: 10.1007/pl00007452. PMID: 9710382.
[2]. Ohbayashi N, Yamagata N, et,al.Enhancement of the structural stability of full-length clostridial collagenase by calcium ions. Appl Environ Microbiol. 2012 Aug;78(16):5839-44. doi: 10.1128/AEM.00808-12. Epub 2012 Jun 8. PMID: 22685155; PMCID: PMC3406112.
[3]. Brandhorst H, Brandhorst D, et,al. Successful human islet isolation utilizing recombinant collagenase. Diabetes. 2003 May;52(5):1143-6. doi: 10.2337/diabetes.52.5.1143. PMID: 12716744.
[4]. Hamzeh Alipour, et al. Therapeutic applications of collagenase (metalloproteases): A review. Asian Pac J Trop Biomed, 2016, 6(11): 975-981.
[5]. O'Flanagan CH, Campbell KR, et,al. Dissociation of solid tumor tissues with cold active protease for single-cell RNA-seq minimizes conserved collagenase-associated stress responses. Genome Biol. 2019 Oct 17;20(1):210. doi: 10.1186/s13059-019-1830-0. PMID: 31623682; PMCID: PMC6796327.
[6]. Kato M, Hattori Y, Kubo M, Maitani Y. Collagenase-1 injection improved tumor distribution and gene expression of cationic lipoplex. Int J Pharm. 2012 Feb 28;423(2):428-34. doi: 10.1016/j.ijpharm.2011.12.015. Epub 2011 Dec 17. PMID: 22197775.