Solubilization, Activation and Partial Purification of a Sialidase from Horse Liver
Using sialyl-methylumbelliferyl -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% T...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Bogor Agricultural University
2005-09-01
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| Series: | Hayati Journal of Biosciences |
| Subjects: | |
| Online Access: | http://journal.ipb.ac.id/index.php/hayati/article/viewFile/180/46 |
| _version_ | 1818578063280046080 |
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| author | KRISHNA PURNAWAN CANDRA PETER ROGGENTIN ROLAND SCHAUER |
| author_facet | KRISHNA PURNAWAN CANDRA PETER ROGGENTIN ROLAND SCHAUER |
| author_sort | KRISHNA PURNAWAN CANDRA |
| collection | DOAJ |
| description | Using sialyl-methylumbelliferyl -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. |
| first_indexed | 2024-12-16T06:39:51Z |
| format | Article |
| id | doaj.art-f0773248359f42338444787b3a41f8d7 |
| institution | Directory Open Access Journal |
| issn | 1978-3019 2086-4094 |
| language | English |
| last_indexed | 2024-12-16T06:39:51Z |
| publishDate | 2005-09-01 |
| publisher | Bogor Agricultural University |
| record_format | Article |
| series | Hayati Journal of Biosciences |
| spelling | doaj.art-f0773248359f42338444787b3a41f8d72022-12-21T22:40:43ZengBogor Agricultural UniversityHayati Journal of Biosciences1978-30192086-40942005-09-011238792Solubilization, Activation and Partial Purification of a Sialidase from Horse LiverKRISHNA PURNAWAN CANDRAPETER ROGGENTINROLAND SCHAUERUsing sialyl-methylumbelliferyl -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.http://journal.ipb.ac.id/index.php/hayati/article/viewFile/180/46SolubilizationActivationPartial PurificationSialidaseHorse Liver |
| spellingShingle | KRISHNA PURNAWAN CANDRA PETER ROGGENTIN ROLAND SCHAUER Solubilization, Activation and Partial Purification of a Sialidase from Horse Liver Hayati Journal of Biosciences Solubilization Activation Partial Purification Sialidase Horse Liver |
| title | Solubilization, Activation and Partial Purification of a Sialidase from Horse Liver |
| title_full | Solubilization, Activation and Partial Purification of a Sialidase from Horse Liver |
| title_fullStr | Solubilization, Activation and Partial Purification of a Sialidase from Horse Liver |
| title_full_unstemmed | Solubilization, Activation and Partial Purification of a Sialidase from Horse Liver |
| title_short | Solubilization, Activation and Partial Purification of a Sialidase from Horse Liver |
| title_sort | solubilization activation and partial purification of a sialidase from horse liver |
| topic | Solubilization Activation Partial Purification Sialidase Horse Liver |
| url | http://journal.ipb.ac.id/index.php/hayati/article/viewFile/180/46 |
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