Ludger Beta-(1-4)-galactosidase enzyme structure

    β(1-4)-galactosidase


    E-BG07

    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.

     View product documentation: Specsheet / CofA / MSDS

    • Part Number
    • E-BG07
    • Amount of Enzyme
    • 0.18 U / 60 µL








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








    Product Specifications:


    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 (E-BG02) 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 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.

    Source: Recombinant Streptococcus pneumoniae in E. coli

    EC: 3.2.1.23

    Alternate Names: β-D-galactoside galactohydrolase, Exo-(1-4)-β-D-galactanase, Lactase

    Contents:
    Ludger Beta-(1-4)-galactosidase - Kit Contents
    β-(1-4) Galactosidase in 20 mM Tris-HCl, 25 mM NaCl (pH 7.5).
    5x Reaction Buffer 6.0 (250 mM sodium phosphate, pH 6.0).

    Specific Activity: >6 U/mg
    Activity: >3 U/mL

    Molecular weight: ~250,000 daltons

    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 5X Reaction Buffer
    4. Add 2 µL β(1-4) Galactosidase
    5. Incubate at 37°C 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.

    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.

    Specific Activity Assay: 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°C pH 5 from p-nitrophenyl-β-D-galactopyranoside.

    Storage: Store enzyme at 4°C.

    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.




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)

Alternative enzymes

  • E-BG02
  • E-AG02
  • β(1-3,4,6)-galactosidase
  • α(1-3,6)-galactosidase





References:

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

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

3. 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)

4. 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.