Abstract
Sialidase secreted by the urease-positiveClostridium sordellii strain G12 was isolated from culture medium and purified to apparent homogeneity as estimated by Fast Protein Liquid Chromatography (FPLC) and sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE). For this purpose, ion-exchange chromatography, gel filtration, isoelectric focusing, and FPLC on ion-exchange resin and gel filtration materials were used. The sialidase was purified 159 300-fold from 5 l of culture medium, yielding 9 μg of enzyme protein with a specific activity of 480 U/mg. For the denatured (SDS-PAGE) and native (FPLC) sialidase relative molecular masses of 40 000 and 38 500 Da, respectively, were estimated. The substrate specificity, kinetic data, and pH-optimum of the enzyme are similar to those of other bacterial sialidases. The influences of salt or serum proteins on enzyme activity are of interest.
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Abbreviations
- MU-Neu5Ac:
-
4-methylumbelliferyl α-d-N-acetylneuraminic acid
- Ganglioside GD1a:
-
IV3NeuAc, ll3NeuAc-GgOse4Cer
- Neu5Ac2en:
-
2-deoxy-2,3-didehydro-N-acetylneuraminic acid
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Roggentin, P., Berg, W. & Schauer, R. Purification and characterization of sialidase fromClostridium sordellii G12. Glycoconjugate J 4, 349–359 (1987). https://doi.org/10.1007/BF01048368
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DOI: https://doi.org/10.1007/BF01048368