| Species : |
Streptococcus pneumoniae |
| Source : |
E. coli |
| Tag : |
Non |
| Description : |
N-acetylglucosaminidase cleaves all non-reducing terminal β-linked N-acetylglucosamine residues from complex carbohydrates and glycoproteins.
The cleavage rates of different linkages of GlcNAc on bi-, tri- and tetraantennary oligosaccharides is greatly dependent on the steric hindrance by neighbouring residues. The β(1-2)GlcNAc residue linked to the β(1-3)-linked mannose is cleaved at the highest rate and the β(1-2) GlcNAc residue linked to the β(1-6)-linked mannose at the lowest rate for all three oligosaccharides. The β(1-6) GlcNAc residue, when present, is removed at the second highest rate and the β(1-4) GlcNAc, third. On a triantennary structure, this residue is removed at the second highest rate. A bisecting β(1-4) GlcNAc linked to the β-linked mannose severely hinders cleavage of other GlcNAc residues–high concentrations of enzymes and prolonged incubation times are required for cleavage. |
| Form : |
Sterile-filtered solution |
| EC : |
3.2.1.52 |
| Specificity : |
Cleaves all non-reducing terminal β-linked N-acetylglucosamine. Bisecting GlcNAc slows the reaction. |
| Contents : |
N-acetylglucosaminidase in 20 mM Tris-HCl, 25 mM NaCl, pH 7.5
5× Reaction Buffer 250 mM sodium phosphate, pH 5.0 |
| Bio-activity : |
Activity: > 50 U/mL
Specific Activity: > 80 U/mg
Defined as the amount of enzyme required to produce 1 μmole of p-nitrophenol (pNP) in 1 minute at 37 centigrade, pH 5.0 from p-nitrophenyl-β-D-N-acetyl-glucosaminide |
| Molecular Mass : |
~140 kDa |
| Suggested usage : |
1. Add up to 100 μg of glycoprotein or 1 nmole of oligosaccharide to a tube.
2. Adjust to 14 μL final volume with de-ionized water.
3. Add 4 μL 5× reaction buffer (pH 5.0).
4. Add 2 μL of N-acetylglucosaminidase.
5. Incubate 3 hours at 37 centigrade.
NOTE: If bisecting GlcNAc or β(1-2)GlcNAc-α(1-6)Man are present increase incubation nation time to 18 hours. |
| Unit Definition : |
One unit of N-acetylglucosaminidase is defined as the amount of enzyme required to produce 1 μmole of p-nitrophenol (pNP) in 1 minute at 37 centigrade, pH 5.0 from p-nitrophenyl-ß-D-N-acetyl-glucosaminide. |
| Purity : |
Each lot of N-acetylglucosaminidase 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. |
| Applications : |
•Structural analysis of oligosaccharides
•Distinguishing different N-acetyl glucosamine link ages
•Distinguishing between N-acetyl glucosamine and N acetylgalactosamine
•Removing heterogeneity from glycoproteins |
| 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. |
| Storage Buffer : |
The enzyme is provided as a sterile-filtered solution in 20 mM Tris-HCl, 25 mM NaCl (pH 7.5). |
