Sialidase Sp α-(2-3)

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).

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Product specification

Contents:

Source: Recombinant Streptococcus pneumoniae in E. coli

EC: 3.2.1.18

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

Sialidase Sp in 50 mM sodium phosphate, pH 7.5
5x Reaction Buffer 250 mM sodium phosphate, pH 6.0

Specific Activity: = 250 U/mg
Activity: = 10 U/mL

Molecular weight: ~75,000 daltons

pH optimum 6.0

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) Sialidase Sp.
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.

Specificity: Cleaves the non-reducing terminal α(2-3) unbranched sialic acid residues from complex carbohydrates and glycoproteins.

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.

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

Applications

• Structural analysis of oligosaccharides
• Determining sialic acid linkage
• Glycoprotein deglycosylation
• Removing heterogeneity from glycoproteins

Recommended Reagents

included with 20 µL and 60 µL pack sizes:
1 vial: Reaction buffer – 400 µl
          250mM Sodium phosphate, pH 6.0

Formulation
The enzyme is provided as a sterile-filtered solution in in 50 mM Sodium phosphate pH 7.5.

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