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Novel methods for the isolation and purification of exoglycosidases
A number of exoglycosidases have been prepared from bacterial and plant sources using established methods for the separation of enzymes, in conjunction with certain novel purification systems hitherto not described in the literature for these enzymes. The enzyme, beta-galactosidase from E. coli has...
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| Format: | Thesis |
| Language: | English |
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Division of Medical Biochemistry and Structural Biology
2017
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| _version_ | 1867613205297102850 |
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| access_status_str | Open Access |
| author | Pannifer, Susan |
| author2 | Merrifield, E H |
| author_browse | Merrifield, E H Pannifer, Susan |
| author_facet | Merrifield, E H Pannifer, Susan |
| author_sort | Pannifer, Susan |
| collection | Thesis |
| description | A number of exoglycosidases have been prepared from bacterial and plant sources using established methods for the separation of enzymes, in conjunction with certain novel purification systems hitherto not described in the literature for these enzymes. The enzyme, beta-galactosidase from E. coli has been prepared using previously described methods of phase separation and ion-exchange chromatography. As a final step in this purification, the use of a new hydroxyl-rich chromatographic support for the isolation of high-grade enzyme suitable for use in enzyme immunoassays was investigated. Methods have also been studied for the recovery of alpha-mannosidase as a by-product of the procedure used for the extraction of urease from jack bean (Canavalia ensiformis). The inclusion of a novel step involving the use of hydrophobic-interaction chromatography on Phenyl-Sepharose led to excellent recoveries of enzyme suitable for commercial use. Studies on a second glycosidase, beta-N-acetylhexosaminidase, from the same source (jack bean) paved the way for an adaptation of existing purification methods to provide increased yields and an improved quality of enzyme. Since the research unit in which this work was performed is associated with commercial organizations responsible for the preparation and marketing of biologically active products, it is important that the methods of purification described in this thesis are compatible with the requirements for largescale purification. |
| format | Thesis |
| id | oai:open.uct.ac.za:11427/26600 |
| institution | University of Cape Town (South Africa) |
| language | eng |
| last_indexed | 2026-06-10T12:32:26.116Z |
| license_str | Not specified — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from UCTD — University of Cape Town Open Access Repository |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| publisher | Division of Medical Biochemistry and Structural Biology |
| publisherStr | Division of Medical Biochemistry and Structural Biology |
| record_format | dspace |
| source_str | UCTD — University of Cape Town Open Access Repository |
| spelling | oai:open.uct.ac.za:11427/26600 Novel methods for the isolation and purification of exoglycosidases Pannifer, Susan Merrifield, E H Glycosidases Glycoside hydrolases - isolation and purification Medical Biochemistry A number of exoglycosidases have been prepared from bacterial and plant sources using established methods for the separation of enzymes, in conjunction with certain novel purification systems hitherto not described in the literature for these enzymes. The enzyme, beta-galactosidase from E. coli has been prepared using previously described methods of phase separation and ion-exchange chromatography. As a final step in this purification, the use of a new hydroxyl-rich chromatographic support for the isolation of high-grade enzyme suitable for use in enzyme immunoassays was investigated. Methods have also been studied for the recovery of alpha-mannosidase as a by-product of the procedure used for the extraction of urease from jack bean (Canavalia ensiformis). The inclusion of a novel step involving the use of hydrophobic-interaction chromatography on Phenyl-Sepharose led to excellent recoveries of enzyme suitable for commercial use. Studies on a second glycosidase, beta-N-acetylhexosaminidase, from the same source (jack bean) paved the way for an adaptation of existing purification methods to provide increased yields and an improved quality of enzyme. Since the research unit in which this work was performed is associated with commercial organizations responsible for the preparation and marketing of biologically active products, it is important that the methods of purification described in this thesis are compatible with the requirements for largescale purification. 2017-12-13T14:15:58Z 2017-12-13T14:15:58Z 1989 Master Thesis Masters MSc (Med) http://hdl.handle.net/11427/26600 eng application/pdf Division of Medical Biochemistry and Structural Biology Faculty of Health Sciences University of Cape Town |
| spellingShingle | Glycosidases Glycoside hydrolases - isolation and purification Medical Biochemistry Pannifer, Susan Novel methods for the isolation and purification of exoglycosidases |
| thesis_degree_str | Master's |
| title | Novel methods for the isolation and purification of exoglycosidases |
| title_full | Novel methods for the isolation and purification of exoglycosidases |
| title_fullStr | Novel methods for the isolation and purification of exoglycosidases |
| title_full_unstemmed | Novel methods for the isolation and purification of exoglycosidases |
| title_short | Novel methods for the isolation and purification of exoglycosidases |
| title_sort | novel methods for the isolation and purification of exoglycosidases |
| topic | Glycosidases Glycoside hydrolases - isolation and purification Medical Biochemistry |
| url | http://hdl.handle.net/11427/26600 |
| work_keys_str_mv | AT pannifersusan novelmethodsfortheisolationandpurificationofexoglycosidases |