Sialidase Cp
Alpha-(2-3,6)

Neuraminidase, N-acetylneuraminate glycohydrolase

Part number E-S005
Source recombinant from Clostridium perfringens in E. Coli.
EC 3.5.1.18

Contents
60 µl aliquot of enzyme (300 mU) in 20 mM Tris-HCl, 25 mM NaCl, pH 7.5
5x Reaction Buffer 250 mM sodium phosphate, pH 6.0

Specific Activity >250 U/mg
Activity 5 U/ml

Molecular weight~41,000 daltons
pH range 4.5-7, optimum 6.0

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 6.0)
3. Add 2 µl of Sialadase Cp
4. Incubate 1 hour at 37°C.

Specifictity Cleaves all nonreducing terminal non-branched alpha-(2-3) and alpha-(2-6)-sialic acid residues from complex carbohydrates and glycoproteins.

Relative cleavage rates for different linkages are: (2-3) > (2-6).

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)-alpha-D-N-acetylneuraminic acid].

Storage Store enzyme at 4°C.

References
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).

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).

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

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).

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

Roggentin, P, B. Rothe, F Lottspeich and R. Schauer. Cloning and sequencing of a Clostridium perfringens sialidase gene. FEBS Lett 238: 31-34 (Se pt 1988).

Roggentin P., R.G . Kleineidam and R. Schauer. Diversity in the properties of two sialidase isoenzymes produced by Clostridium perfringens spp. Biol Chem Hoppe-Seyler 376: 569-575 (1995)