| Species : |
Streptococcus pneumoniae |
| Source : |
E. coli |
| Tag : |
Non |
| Description : |
Non-reducing terminal β(1-4)-Galactose. The number of antennae does not affect cleavage rate. Fucose linked to the penultimate N-acetylglucosamine will block the cleavage of the galactose.
β Galactosidase from Streptococcus pneumoniae releases only β(1-4) linked, non-reducing terminal galactose from complex carbohydrates and glycoproteins. β(1-4) galactose is by far the most common linkage found in N-linked oligosaccharides. For other galactosidase linkages, β(1-3,4,6)-Galactosidase from Bovine testes is recommended. The enzyme is as active on tetraantennary oligosaccharides as on disaccharides containing β(1-4)-linked galactose. Fucose linked to the penultimate N-acetylglucosamine will block the cleavage of the galactose. Up to 100 μg of asialofetuin can be completely β(1-4)-degalactosylated in less than 1 hour using 3 mU of enzyme. |
| Form : |
Sterile-filtered solution |
| EC : |
3.2.1.23 |
| Specificity : |
Non-reducing terminal β(1-4)-galactose. Number of antennae does not affect cleavage rate. Fucose linked to the penultimate N-acetylglucosamine will block cleavage of the galactose. |
| Contents : |
β-(1-4) Galactosidase in 20 mM Tris-HCl, 25 mM NaCl (pH 7.5).
5× Reaction Buffer 6.0 (250 mM sodium phosphate, pH 6.0). |
| Bio-activity : |
Specific Activity: > 6 U/mg
Activity: > 3 U/mL |
| Molecular Mass : |
~250 kDa |
| Suggested usage : |
1. Add up to 100 μg of asialoglycoprotein or 1 nmol of oligosaccharide to tube.
2. Add water to 14 μL
3. Add 4 μL 5× Reaction Buffer
4. Add 2 μL β(1-4) Galactosidase
5. Incubate at 37 centigrade for 1 hour.
For glycoproteins, cleavage may be monitored by SDS-PAGE if the size differential between native and de-galactosylated protein is sufficient for detection.
Note: The optimum for cleavage of oligosaccharides is ~ pH 6.0. |
| Unit Definition : |
One unit of β Galactosidase is defined as the amount of enzyme required to produce 1 μmole of p-nitrophenol (pNP) in 1 minute at 37 centigrade pH 5 from p-nitrophenyl-β-D-galactopyranoside. |
| Purity : |
Each lot of β Galactosidase is tested for contaminating protease as follows; 10 μg of denatured BSA is incubated for 24 hours with 2 μL of enzyme. SDS-PAGE analysis of the treated BSA shows no evidence of degradation.
The production host strain has been extensively tested and does not produce any detectable glycosidases. |
| Stability : |
Stable at least 12 months when stored properly. Several days exposure to ambient temperatures will not reduce activity. |
| Storage : |
Store enzyme at 4 centigrade. Do not freeze. |
| Storage Buffer : |
The enzyme is provided as a sterile-filtered solution in 20 mM Tris-HCl, 25 mM NaCl (pH 7.5). |
