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Title: | Solubilization, Activation and Partial Purification of a Sialidase from Horse Liver |
Other Titles: | HAYATI Journal of Biosciences Vol. 12 No.3 Tahun 2005 |
Authors: | Candra, Krishna Purnawan Roggentin, Peter Schauer, Roland |
Issue Date: | 2005 |
Publisher: | IPB (Bogor Agricultural University) |
Series/Report no.: | Vol 12;No 3 |
Abstract: | Using sialyl-methylumbelliferyl α-glycoside as substrate, sialidase in horse liver was detected as a membrane-bound enzyme. A yield of about 50% of sialidase activity was found in supernatant when solubilized in 0.1 M sodium-phosphate buffer pH 5.5, containing 0.15 M NaCl, 0.25 M sucrose, and 0.5% Triton X-100. Sialidase in the solubilisate could be activated by incubating in acidic pH at 37 oC. Incubation of this solubilized enzyme at 37 oC for 1.5 h at pH 5.0 led to 10% increase of activity and to the precipitation of about 50% of contaminating protein. Using cation-exchange chromatography on S-Sepharose FF and affinity chromatography on p-aminophenyl oxamic acid-agarose following solubilization and activation, about 6% of total sialidase activity was recovered with the purification factor of about 500. The pH and temperature optimum were measured at pH 4.3 and between 37-45 oC, respectively. Neu5Ac2en was a strong inhibitor, while p-aminophenyl oxamic acid had only a weak inhibitory effect. |
URI: | http://repository.ipb.ac.id/handle/123456789/43379 |
ISSN: | 0854-8587 |
Appears in Collections: | Hayati Journal of Biosciences |
Files in This Item:
File | Description | Size | Format | |
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Krishna Purnawan Candra.pdf | e-Journal | 296.51 kB | Adobe PDF | View/Open |
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