Neuraminidase, N-acetylneuraminate glycohydrolase
Part number E-S007
Source recombinant Streptococcus pneumoniae in E. Coli
60 µl aliquot of enzyme (300 mU) 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
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 Sp
4. Incubate 1 hour at 37˚C.
Specifictity Cleaves the non-reducing terminal alpha-(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)-alpha-D-N-acetylneuraminic acid].
Storage Store enzyme at 4˚C.
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).
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).
Kelly, R. T., D. Greiff and S. Farmer. Neuraminidase activity in Diplococcus pneumoniae. J Bacteriol 91: 601-3 (1965).
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).