Ludger beta-(1-2,3,4,6)-n-acetylglucosaminidase enzyme structure

    β(1-2,3,4,6)-N-acetylglucosaminidase


    E-GL01

    N-acetylglucosaminidase cleaves all non-reducing terminal β-linked N-acetylglucosamine residues from complex carbohydrates and glycoproteins.

     View product documentation: Specsheet / CofA / MSDS

    • Part Number
    • E-GL01
    • Amount of Enzyme
    • 2.4 U / 60 µL








    Kit includes enzyme plus reaction buffer.
    Sufficient for up to 60 reactions.






















    Product Specifications:


    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 neighboring 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.

    Source: Recombinant from Streptococcus pneumoniae in E. Coli.

    EC: 3.2.1.52

    Alternate Names: β-N-acetylglucosaminidase, N-acetyl-β-d-glycosaminide N-acetylglucosaminohydrolase, glucosaminidase, hexosaminidase

    Contents:
    Ludger beta-(1-2,3,4,6)-n-acetylglucosaminidase - Kit Contents
    N-acetylglucosaminidase in 20 mM Tris-HCl, 25 mM NaCl, pH 7.5
    5x Reaction Buffer 250 mM sodium phosphate, pH 5.0

    Specific Activity: >80 U/mg
    Activity: >50 U/mL

    Molecular weight: ~140,000 daltons

    pH range: 5-7, optimum 5.0

    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 5x reaction buffer (pH 5.0).
    4. Add 2 µL of N-acetylglucosaminidase.
    5. Incubate 3 hours at 37°C.

    NOTE: If bisecting GlcNAc or β(1-2)GlcNAc-α(1-6)Man are present increase incubation nation time to 18 hours.

    Specificity: Cleaves all non-reducing terminal β-linked N-acetylglucosamine. Bisecting GlcNAc slows the reaction.

    Specific Activity Assay: Defined as the amount of enzyme required to produce 1 µmole of p-nitrophenol (pNP) in 1 minute at 37°C, pH 5.0 from p-nitrophenyl-β-D-N-acetyl-glucosaminide

    Storage: Store enzyme at 4°C.

    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.

    Stability: Stable at least 12 months when stored properly. Several days exposure to ambient temperatures will not reduce activity.





Companion Products:

Process controls - View our N-glycan Nomenclature Table


Glycan clean-up - removal of enzymes from glycan mixtures following exoglycosidase treatment

  • LC-EXO-A6
  • LC-EXO-96
  • LudgerClean Post-Exoglycosidase Clean-up Spin Columns (6 samples)
  • LudgerClean Post-Exoglycosidase Clean-up Plate (96 samples)

Labelling and derivatisation of released glycans - View our Glycan Labelling Summary Table

  • LT-KAB-A2
  • LT-KAB-VP24
  • LT-KAB-VP96
  • LT-KPROC-24
  • LT-KPROC-96
  • LT-KPROC-VP24
  • LT-KAA-A2
  • LT-KAA-VP24
  • LT-PERMET-96
  • LT-PERMET-VP96
  • LudgerTag 2-AB Glycan Labelling Kit, sodium cyanoborohydride (24 samples)
  • LudgerTag 2-AB Glycan Labelling Kit, picoline borane (24 samples)
  • LudgerTag 2-AB Glycan Labelling Kit, picoline borane (96 samples)
  • LudgerTag Procainamide Glycan Labelling Kit, sodium cyanoborohydride (24 samples)
  • LudgerTag Procainamide Glycan Labelling Kit, sodium cyanoborohydride (96 samples)
  • LudgerTag Procainamide Glycan Labelling Kit, picoline borane (24 samples)
  • LudgerTag 2-AA Glycan Labelling Kit, sodium cyanoborohydride (24 samples)
  • LudgerTag 2-AA Glycan Labelling Kit, picoline borane (24 samples)
  • LudgerTag Permethylation Kit (96 samples)
  • LudgerTag Permethylation Kit, without methyl iodide (96 samples)






References:

1. Clarke, V. A., N. Platt and T.D. Betters. Cloning and expression of the beta-N-acetylglucosaminidase gene from Steptococcus pneumoniae. Generation of truncated enzymes with modified aglyconn specificity. J Biol Chem 270:8805-8814 (1995).

2. Dwek, R. A., C. J. Edge, D. J. Harvey, M. R. Wormald and R. B. Parekh. Analysis of glycoprotein-associated oligosaccharides. Ann Rev Biochem 62: 65-100 (1993).

3. Glasgow, L.R., J.C. Paulson and R.L. Hill. Systematic purification of five glycosidases from Streptococcus pneumoniae J Biol Chem 252:8615-8623(1977).

4. Kobata, A. Use of endo- and exoglycosidases for structural studies of glycoconjugates. Anal Biochem 100: 1-14 (1979).

5. Prime, S., J. Dearnley , A. M. Venton, R. B. Parekh and C. J. Edge. Oligosaccharide sequencing based on exo- and endoglycosidase digestion and liquid chromatographic analysis of the products. J Chromatogr A 720: 263-274 (1996).