Purification and partial characterization of superoxide dismutase from the thermophilic bacteria Thermothrix sp
Superoxide dismutase (SOD; EC 1.15.1.1), a high molecular weight component of the antioxidant defense system, provided promising results in the treatment of oxidative damage. Thermothrix sp., isolated from thermal spa water in Serbia, showed high superoxide dismutase activity. The SOD, from cell fre...
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| Format: | Article |
| Language: | English |
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Serbian Chemical Society
2004-01-01
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| Series: | Journal of the Serbian Chemical Society |
| Subjects: | |
| Online Access: | http://www.doiserbia.nb.rs/img/doi/0352-5139/2004/0352-51390401009S.pdf |
| _version_ | 1818293192439627776 |
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| author | Šeatović Svetlana Gligić Ljubinka Radulović Željka Jankov Ratko M. |
| author_facet | Šeatović Svetlana Gligić Ljubinka Radulović Željka Jankov Ratko M. |
| author_sort | Šeatović Svetlana |
| collection | DOAJ |
| description | Superoxide dismutase (SOD; EC 1.15.1.1), a high molecular weight component of the antioxidant defense system, provided promising results in the treatment of oxidative damage. Thermothrix sp., isolated from thermal spa water in Serbia, showed high superoxide dismutase activity. The SOD, from cell free extract, was purified to homogenity by ammonium sulfate precipitation, Sephadex G 75 gel filtration chromatography and QAE Sephadex ion exchange chromatography. The specific activity of the purified enzyme was 9191 U/mg. The purified enzyme was analyzed and partially characterized. SOD was localized in polyacrylamide gel by activity staining, based on the reduction of nitroblue tetrazolium (NBT) by superoxide. The enzyme molecular weight determined by gel chromatography is 37 kD. According to SDS PAGE it is composed of two subunits of equal size, joined by noncovalent interactions. The isoelectric point, assessed by isoelectric focusing is 5.3. The optimum pH for enzyme activity was in the range of 8 to 10. The optimum temperature for SOD activity was 60 ºC. After one hour of incubation at 40, 50 and 60 ºC the SOD activity increases, but at 80 ºC, the SOD is denaturated. After 24 hours of incubation at 25 ºC SO Dactivity only slightly decreases. |
| first_indexed | 2024-12-13T03:11:57Z |
| format | Article |
| id | doaj.art-aaf98bb393a3441ab421264ccc2b26f0 |
| institution | Directory Open Access Journal |
| issn | 0352-5139 1820-7421 |
| language | English |
| last_indexed | 2024-12-13T03:11:57Z |
| publishDate | 2004-01-01 |
| publisher | Serbian Chemical Society |
| record_format | Article |
| series | Journal of the Serbian Chemical Society |
| spelling | doaj.art-aaf98bb393a3441ab421264ccc2b26f02022-12-22T00:01:35ZengSerbian Chemical SocietyJournal of the Serbian Chemical Society0352-51391820-74212004-01-0169191610.2298/JSC0401009S0352-51390401009SPurification and partial characterization of superoxide dismutase from the thermophilic bacteria Thermothrix spŠeatović Svetlana0Gligić Ljubinka1Radulović Željka2Jankov Ratko M.3Galenika a.d., Institute, Belgrade, Serbia and MontenegroGalenika a.d., Institute, Belgrade, Serbia and MontenegroGalenika a.d., Institute, Belgrade, Serbia and MontenegroFaculty of Chemistry, University of Belgrade, Belgrade, Serbia and MontenegroSuperoxide dismutase (SOD; EC 1.15.1.1), a high molecular weight component of the antioxidant defense system, provided promising results in the treatment of oxidative damage. Thermothrix sp., isolated from thermal spa water in Serbia, showed high superoxide dismutase activity. The SOD, from cell free extract, was purified to homogenity by ammonium sulfate precipitation, Sephadex G 75 gel filtration chromatography and QAE Sephadex ion exchange chromatography. The specific activity of the purified enzyme was 9191 U/mg. The purified enzyme was analyzed and partially characterized. SOD was localized in polyacrylamide gel by activity staining, based on the reduction of nitroblue tetrazolium (NBT) by superoxide. The enzyme molecular weight determined by gel chromatography is 37 kD. According to SDS PAGE it is composed of two subunits of equal size, joined by noncovalent interactions. The isoelectric point, assessed by isoelectric focusing is 5.3. The optimum pH for enzyme activity was in the range of 8 to 10. The optimum temperature for SOD activity was 60 ºC. After one hour of incubation at 40, 50 and 60 ºC the SOD activity increases, but at 80 ºC, the SOD is denaturated. After 24 hours of incubation at 25 ºC SO Dactivity only slightly decreases.http://www.doiserbia.nb.rs/img/doi/0352-5139/2004/0352-51390401009S.pdfsuperoxide dismutasethermothrix sp.isolationpurificationcharacterization |
| spellingShingle | Šeatović Svetlana Gligić Ljubinka Radulović Željka Jankov Ratko M. Purification and partial characterization of superoxide dismutase from the thermophilic bacteria Thermothrix sp Journal of the Serbian Chemical Society superoxide dismutase thermothrix sp. isolation purification characterization |
| title | Purification and partial characterization of superoxide dismutase from the thermophilic bacteria Thermothrix sp |
| title_full | Purification and partial characterization of superoxide dismutase from the thermophilic bacteria Thermothrix sp |
| title_fullStr | Purification and partial characterization of superoxide dismutase from the thermophilic bacteria Thermothrix sp |
| title_full_unstemmed | Purification and partial characterization of superoxide dismutase from the thermophilic bacteria Thermothrix sp |
| title_short | Purification and partial characterization of superoxide dismutase from the thermophilic bacteria Thermothrix sp |
| title_sort | purification and partial characterization of superoxide dismutase from the thermophilic bacteria thermothrix sp |
| topic | superoxide dismutase thermothrix sp. isolation purification characterization |
| url | http://www.doiserbia.nb.rs/img/doi/0352-5139/2004/0352-51390401009S.pdf |
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