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C9263

Sigma-Aldrich

Collagenase from Clostridium histolyticum

Type V, ≥1 FALGPA units/mg solid, >125 CDU/mg solid

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Synonym(s):
Clostridiopeptidase A
CAS Number:
9001-12-1
Enzyme Commission number:
3.4.24.3 (BRENDA, IUBMB)
EC Number:
232-582-9
MDL number:
MFCD00130830
NACRES:
NA.54

Quality Level

200

form

powder

specific activity

≥1 FALGPA units/mg solid
>125 CDU/mg solid

mol wt

113-114 kDa

solubility

TESCA buffer (50 mM TES, 0.36 mM Calcium chloride, pH 7.4): soluble 1-2 mg/mL

application(s)

diagnostic assay manufacturing

storage temp.

−20°C

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General description

Clostridium histolyticum collagenases (CCH) are classified into two types ; I and II. Both are a single polypeptide with molecular weight of 114kDa - 113kDa, respectively. CCH I contains tandem domains and CCH II contain single domain for collagen binding.

Application

Collagenase from Clostridium histolyticum has been used in a study to investigate the degradation of collagen by the cariogenic bacteria, Streptococcus mutans. Collagenase from Clostridium histolyticum has also been used in a study to investigate the survivability of collagen micronetworks in the presence of collagenase.
Collagenase has been used in the preparation of arterial tissue for the study of Advanced Glycosylation End Products (AGE). The enzyme has also been used along with other proteases for the disaggregation of human tumor, mouse kidney, human brain, and lung epithelial tissues among several others. It is also effective in liver and kidney perfusion studies, digestion of pancreas, and isolation of nonparenchymal hepatocytes. The product has been used to digest forearm skin biopsies from Parkinson disease patients to grow human skin fibroblast primary cultures. The enzyme from Sigma has been used for rodent islet isolation in a study that assayed glucokinase functions.

Biochem/physiol Actions

Clostridium histolyticum collagenase (CCH) is a digestive enzyme acting particularly on collagen. It is useful in the treatment of Dupuytren′s contracture, fibroproliferative disorder.
Collagenase is activated by four gram atom calcium per mole enzyme. It is inhibited by ethylene glycol-bis(beta-aminoethyl ether) - N, N, N′,N′-tetraacetic acid, beta-mercaptoethanol, glutathione, thioglycolic acid and 8-hydroxyquinoline.

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.

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substrate

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Pictograms

Health hazard
GHS08

Signal Word

Danger

Hazard Statements

H334

Precautionary Statements

P261 - P284 - P501

Hazard Classifications

Resp. Sens. 1

Storage Class Code

11 - Combustible Solids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Regulatory Information

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Collagenase Clostridium histolyticum: emerging practice patterns and treatment advances
Warwick D, et al.
Journal of Plastic Surgery and Hand Surgery, 50(5), 251-261 (2016)
Ali Jazayeri et al.
Nature, 546(7657), 254-258 (2017-06-01)
Glucagon-like peptide 1 (GLP-1) regulates glucose homeostasis through the control of insulin release from the pancreas. GLP-1 peptide agonists are efficacious drugs for the treatment of diabetes. To gain insight into the molecular mechanism of action of GLP-1 peptides, here
Agnes Petit et al.
The Journal of biological chemistry, 280(40), 34025-34032 (2005-08-05)
Mutations in the PTEN-induced kinase 1 (PINK1) gene have recently been implicated in autosomal recessive early onset Parkinson Disease (1, 2). To investigate the role of PINK1 in neurodegeneration, we designed human and murine neuronal cell lines expressing either wild-type
Nicolai M Doliba et al.
American journal of physiology. Endocrinology and metabolism, 302(1), E87-E102 (2011-09-29)
It was reported previously that isolated human islets from individuals with type 2 diabetes mellitus (T2DM) show reduced glucose-stimulated insulin release. To assess the possibility that impaired bioenergetics may contribute to this defect, glucose-stimulated respiration (Vo(2)), glucose usage and oxidation
Daniela Frasca et al.
Frontiers in immunology, 11, 392-392 (2020-03-19)
We have previously shown that the human obese adipose tissue (AT) contributes to increased secretion of adipocyte-specific IgG antibodies in individuals with obesity. This occurs without any exogenous stimulation, because the ongoing process of cell death in the obese AT
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Protocols

Enzymatic Assay of Collagenase (EC 3.4.24.3) using FALGPA (N-(3-[2-Furyl]acryloyl)-Leu-Gly-Pro-Ala)

To measure collagenase activity, N-(3-[2-Furyl]acryloyl)-Leu-Gly-Pro-Ala is used in a continuous spectrophotometric rate determination at 345 nm. Collagenase hydrolyzes collagen peptide bonds.

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