Purification and characterization of sialidase fromClostridium sordellii G12 |
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Authors: | Peter Roggentin Wigbert Berg Roland Schauer |
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Institution: | (1) Biochemisches Institut, Christian-Albrechts-Universität, Olshausenstr. 40, D-2300 Kiel, Federal Republic of Germany |
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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 g 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.Abbreviations MU-Neu5Ac
4-methylumbelliferyl -d-N-acetylneuraminic acid
- Ganglioside GD1a
IV3NeuAc, ll3NeuAc-GgOse4Cer
- Neu5Ac2en
2-deoxy-2,3-didehydro-N-acetylneuraminic acid |
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Keywords: | sialidase (neuraminidase) purification properties Clostridium sordellii |
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