| Species : |
Bacteroides fragilis |
| Source : |
E. coli |
| Tag : |
Non |
| Description : |
Endo-β-Galactosidase cleaves internal β(1-4) galactose linkages in unbranched, repeating poly-N-acetyllactosamine structures. Sulfated structures such as keratan sulfate are also cleaved. Branching and/or fucosylation of the substrate may decrease or eliminate cleavage. |
| Form : |
Sterile-filtered solution |
| EC : |
3.2.1.103 |
| Specificity : |
Cleaves internal β(1-4) galactose linkages in unbranched, repeating poly-N-acetyllactosamine [GlcNAc-β(1-3)Gal-β(1-4)]n structures are the preferred substrate. Sulfated structures such as keratan sulfate are also cleaved. Branching and/or fucosylation of the substrate may decrease or eliminate cleavage. Sulfation of C-6 on galactose will block cleavage. Oligosaccharidesof the neo-lacto group are cleaved at greatly educed rates depending on the deviation from the preferred substrate. For example, Gal-β(1-3)GlcNAc-β(1-3)Gal-β(1-4)Glc is cleaved at 5×10-5 the rate of keratan sulfate. Specificity is similar to the Escherichia freundii enzyme except that it is limited to cleaving N-acetyllactosamine extensions on tetraantennary structures of erythropoietin. |
| Contents : |
60 μL aliquot of enzyme (0.9 U) in 20 mM tris-HCl, pH 7.5
1 vial reaction buffer- 250mM Sodium phosphate, pH 5.8 |
| Bio-activity : |
Specific Activity: > 140 U/mg
Activity: > 14 U/mL |
| Molecular Mass : |
~32 kDa |
| Suggested usage : |
For glycoproteins:
1. Add up to 100 μg of glycoprotein to a tube.
2. Add 4 μL 5× buffer and water to 19 μL.
3. Add 1 μL enzyme.
4. Incubate at 37 centigrade for 2 hrs. |
| Unit Definition : |
One unit of Endo-β-Galactosidase is defined as the amount that will liberate one μmole of reducing sugar per minute at 37 centigrade and pH 5.8 from bovine corneal keratan sulfate. |
| pH optimum : |
5.8 |
| Purity : |
Endo-β-Galactosidase is tested for contaminating protease as follows: 10 μg of denatured BSA is incubated for 24 hours at 37 centigrade with 2 μL of enzyme. SDS-PAGE analysis of the treated BSA shows no evidence of degradation.
The production strain of E. coli has been extensively tested and does not produce any detectable glycosidases. |
| Applications : |
Endo-β-Galactosidase (EC 3.2.1.103) cleaves internal β(1-4) galactose linkages in unbranched, repeating polyN-acetyllactosamine structures. Sulfated structures such as keratan sulfate are also cleaved. Branching and/or fucosylation of the substrate may decrease or eliminate cleavage.
Endo-β-Galactosidase is useful for identifying and removing poly-N-acetyllactosamine structures on many biologically important glycoconjugates. |
| Stability : |
Stable at least 24 months when stored properly. Several days exposure to ambient temperature will not reduce activity. Active for at least 5 days under reaction conditions. |
| Storage : |
Store enzyme at 4 centigrade. |
| Storage Buffer : |
The enzyme is provided as a sterile-filtered solution in 20 mM Tris-HCl, pH 7.5 |
