性能描述

性状： 基本上无盐，冻干粉
保存温度  -20°C 

产品说明：
“Crude” collagenase refers to the material purified from the fermentation of Clostridium histolyticum bacteria. It is actually a mixture of sev eral different enzymes including collagenase, which act together to break down tissue. This preparation contains collagenase, non-specific proteases, clostripain, neutral protease, and aminopeptidase activities. Crude collagenase is equivalent to the first 40% ammonium sulfate fraction
prepared by Mandl.
Molecular mass (SDS-PAGE):68–130 kDa
As many as seven collagenase proteins can be
present, some of these are C-truncated forms of the
Type I and Type II collagenases (sometimes called
collagenases A and B) that are expressed from two
genes, colG and colH.Molecular mass (sequence): The colG and colH genes
have been isolated from C. histolyticum JCM 1403
(ATCC 19401).The colG gene encodes a mature 113,897 Da Type I collagenase and several C-truncated forms. The colH gene encodes the mature 111,963 Dal Type II collagenase and one C-truncated
form.Optimal pH:6.3–8.8 The enzyme is assayed at pH 7.4
Isoelectric Point (pI): Six collagenases purified from
crude collagenase had isoelectric points in the range of
5.35–6.20.Specificity: collagenase recognizes the following
peptide sequence where X is most often a neutral
amino acid:–R–Pro–X–Gly–Pro–R–
Cleavage site Substrates: In addition to the various natural collagen
substrates, many synthetic substrates have been
prepared:
Z-Gly-Pro-Gly-Gly-Pro-Ala (Catalog Numbers 27673
and 27670; KM = 0.71 mM)
Z-Gly-Pro-Leu-Gly-Pro
N-2,4-Dinitrophenyl-Pro-Gln-Gly-Ile-Ala-Gly-Gln-D-Arg
N-(3-(2-furyl)acryloyl)-Leu-Gly-Pro-Ala
4-Phenylazobenzoxycarbonyl-Pro-Leu-Gly-Pro-D-Arg
N-succinyl-Gly-Pro-Leu-Gly-Pro 7-amido-4-methyl-coumarin (substrate for “collagenase-like peptidase”)N-(2,4-Dinitrophenyl)-Pro-Leu-Gly-Leu-Trp-Ala-D-Arg amide (substrate for “vertebrate collagenase”)

Activators/Cofactors: Collagenase activity is stabilized
by 0.1 mole calcium ions (Ca2+) per mole of enzyme.Calcium ions also facilitate binding to the collagen Molecule.Zinc ions (Zn2+) are required for activity, but are tightly bound to the collagenase during purification.Additional Zn2+
should not be necessary
as long as no chelator is added during digestion.
Inhibitors:
Ethylene glycol-bis(2-aminoethylether)-N,N,N¢,N¢-tetra-acetic acid
Ethylenediaminetetraacetic acid
b-Thujaplicin (Hinokitiol)
Cysteine
2-mercaptoethanol (Catalog Number M7522)
Glutathione (reduced)
Thioglycolic acid, sodium
8-Hydroxyquinoline-5-sulfonate

Application
Collagenase products offered by Sigma-Aldrich are prepared from culture supernatants of C. histolyticum. Most preparations contain collagenase, non-specifc protease, and clostripain activities. These enzymes act synergistically to degrade collagen and other intercellular material. Crude Collagenases are suitable for isolation of cells from many types of animal tissue. All are evaluated for their enzymatic activities. The optimal concentration for use and digestion time of tissue needs to be empirically determined. In addition, Sigma-Aldrich also offers collagenases which are tested for suitability for release of cells from specified tissues (release of rat pancreatic islets, rat epididymal adipocytes, and rat hepatocytes.

Clostridiopeptidase A has been used in a study to investigate the quantitative analysis of denatured collagen by collagenase digestion and subsequent MALDI-TOF mass spectrometry. Clostridiopeptidase A has also been used in a study to investigate the role of single-dose clostridiopeptidase A collagenase in peritoneal adhesions.

Unit Definition
One FALGPA unit hydrolyzes 1.0 μmole of FALGPA (Furylacryloyl-LEU-GLY-PRO-ALA, F5135) per minute at pH 7.5 at 25 °C in the presence of calcium ions.

One collagen digestion unit (CDU) liberates peptides from collagen (C9879) 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.