Neuraminidase, N-acetylneuraminate glycohydrolase
Part number E-S001
Source recombinant from Arthrobacter ureafaciens in E. Coli.
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 >135 U/mg
Activity 5 U/ml
Molecular weight ~69,000 daltons
pH range 4.5-7, optimum 6.0
add 2 µL of enzyme to 100 µg of glycoprotein or 1 nmol of oligosaccharide. Add buffer and incubate 3 hours at 37˚C. NOTE: longer incubation times are necessary if branched sialic acids are present.
Specifictity 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)-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).
Ohta, Y., Y. Tsukada and T. Sugimori. Purification and properties of neuraminidase isoenzymes in Arthrobacter ureafaciens mutant. J Biochem (Tokyo) 106: 1086- 1089 (1989).
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).