Ludger Sialidase Au enzyme structure

    Sialidase Au α(2-3,6,8,9)


    E-S001

    α(2-3,6,8,9) Sialidase cleaves all non-reducing terminal sialic acid residues from complex carbohydrates and glycoproteins. In addition, the enzyme will cleave branched sialic acids (linked to an internal residue).

     View product documentation: Specsheet / CofA / MSDS

    • Part Number
    • E-S001
    • Amount of Enzyme
    • 0.3 U / 60 µL








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







    Product Specifications:


    α(2-3,6,8,9) Sialidase Au cleaves all cleaves all non-reducing terminal sialic acid residues from complex carbohydrates and glycoproteins. The relative cleavage rates for different linkages are: α(2-6) > α(2-3) > α(2-8), α(2-9).

    In addition, the enzyme will cleave branched sialic acids (linked to an internal residue). This property makes it unique among sialidases. High concentrations of enzymes and prolonged incubation times may be required for cleaving branched residues. To cleave only non- reducing terminal α(2-3) unbranched sialic acid residues, use Sialidase SP, part number E-S007.

    α(2-3,6,8,9) Sialidase Au is isolated from a clone of Arthrobacter ureafaciens. The enzyme has been extensively characterized using oligosaccharide standards.

    Source: Recombinant from Arthrobacter ureafaciens in E. Coli.

    EC: 3.2.1.18

    Alternate Names: Neuraminidase, NANase, N-acetylneuraminate glycohydrolase, Exo-α-sialidase

    Contents:
    Ludger Sialidase Au - Kit Contents
    Sialidase Au in 20 mM Tris-HCl, 25 mM NaCl, pH 7.5
    5x Reaction Buffer 250 mM sodium phosphate, pH 6.0

    Specific Activity: >135 U/mg
    Activity: >5 U/mL

    Molecular weight: 70,000 daltons

    pH range: 4.5-7, optimum 6.0
    The recommended buffer concentrate provides the optimal pH for enzyme activity with the standard substrate. If glycosidase treatment is performed at suboptimal pH because of glycoprotein solubility or activity requirements, expect some diminution in enzyme activity.

    Suggested usage:
    1. Add up to 100 µg of glycoprotein or 1 nmol of oligosaccharide to tube.
    2. Add water to 14 µL
    3. Add 4 µL 5X Reaction Buffer.
    4. Add 2 µL α(2-3,6,8,9) Sialidase Au.
    5. Incubate at 37°C for 1 hour.

    Desialylation may be monitored by SDS-PAGE if the size differential between native and desialylated protein is sufficient for detection.

    NOTE: longer incubation times are necessary if branched sialic acids are present.

    Specificity: Cleaves all non-reducing terminal branched and unbranched sialic acids

    Specific Activity: Defined as the amount of enzyme required to produce 1 µmole of methylumbelliferone in 1 minute at 37°C, pH 5.0 from MU-NANA [2′-(4-methylumbelliferyl)-α-D-N-acetylneuraminic acid].

    Storage: Store enzyme at 4°C.

    Purity: Each lot of α(2-3,6,8,9) Sialidase Au 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. Enzyme remains active at 37°C for at least one week.





Companion Products:

Process controls - View our N-glycan Nomenclature Table

  • CAB-STP-NEUAC-01
  • CPROC-STP-NEUAC-01
  • Sialidase Test Panel - mix of 2-AB labelled glycans with different NeuAc linkages *Coming Soon
  • Sialidase Test Panel - mix of procainamide labelled glycans with different NeuAc linkages *Coming Soon

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-S005
  • E-S007
  • LZ-ACASE-KIT
  • Sialidase Cp α(2-3,6)
  • Sialidase Sp α(2-3)
  • Acetyl esterase (sialate-O-acetylesterase), removal of O-acetyl groups from sialic acids






References:


1. Corfield, A. P., H. Higa, J. C. Paulson and R. Schauer. The specificity of viral and bacterial sialidases for alpha(2-3) and alpha(2-6)-linked sialic acids in glycoproteins. Biochim Biophys Acta 744: 121-12 6 (1983).

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

4. Ohta, Y., Y. Tsukada and T. Sugimori. Purification and properties of neuraminidase isoenzymes in Arthrobacter ureafaciens mutant. J Biochem (Tokyo) 106: 1086- 1089 (1989).

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).

6. Uchida, Y., Y. Tsukada and T. Sugimori. Enzymatic properties of neuraminidases from Arthrobacter ureafaciens. J Biochem (Tokyo) 86: 573-58 5 (1979).