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Preparation of a self-contained NADH co-factor recycling particle system
Includes abstract.
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| Main Author: | |
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| Other Authors: | |
| Format: | Thesis |
| Language: | English |
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Department of Molecular and Cell Biology
2014
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| _version_ | 1867613168186949633 |
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| access_status_str | Open Access |
| author | Twala, Busisiwe V |
| author2 | Sewell, Bryan Trevor |
| author_browse | Sewell, Bryan Trevor Twala, Busisiwe V |
| author_facet | Sewell, Bryan Trevor Twala, Busisiwe V |
| author_sort | Twala, Busisiwe V |
| collection | Thesis |
| description | Includes abstract. |
| format | Thesis |
| id | oai:open.uct.ac.za:11427/4343 |
| institution | University of Cape Town (South Africa) |
| language | eng |
| last_indexed | 2026-06-10T12:31:50.330Z |
| license_str | Not specified — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from UCTD — University of Cape Town Open Access Repository |
| publishDate | 2014 |
| publishDateRange | 2014 |
| publishDateSort | 2014 |
| publisher | Department of Molecular and Cell Biology |
| publisherStr | Department of Molecular and Cell Biology |
| record_format | dspace |
| source_str | UCTD — University of Cape Town Open Access Repository |
| spelling | oai:open.uct.ac.za:11427/4343 Preparation of a self-contained NADH co-factor recycling particle system Twala, Busisiwe V Sewell, Bryan Trevor Jordaan, Justin Cell Biology Includes abstract. Includes bibliographical references (leaves 101-116). Oxidoreductases are interesting enzymes with potential applications in a number of different industries such as the textile, food and feed, chemical and biomedical industries. Oxidoreductases require the use of co-factors. These small molecules are relatively expensive and are required in stoichiometric amounts for their enzymatic reaction; this negatively impacts the economic viability of their potential applications. Several methods have been developed to counteract this problem, the most preferred of which is the enzymatic co-factor recycling method. A few methods for the co-immobilisation of enzymes and co-factors have been developed. These systems are of interest as they offer the advantages of recycling the enzymes together with the co-factor, thereby enabling re-use. The immobilisation of enzymes also provides a platform for improving their stability, activity, specificity and selectivity. Since glucose dehydrogenase (GDH) and NADH oxidase, are industrially relevant co-factor recycling enzymes for NAD(P)H and NAD+ respectively, characterisation of their immobilisation is of interest. The current work describes the use of the proprietary particle technology, termed ReSyn™, for the construction of a self-contained co-factor recycling system. The research included the optimisation of immobilisation for the individual enzymes, followed by the co-immobilisation with subsequent co-factor entrapment. The immobilised enzymes displayed improved thermal and pH stability compared to the non-immobilised enzymes. Immobilised GDH also displayed increased activity over the acidic range when compared to free GDH. The system was shown to be capable of recycling NADH/NAD+ up to at least 142 times with a specific activity of 10.18 U.mg¯1. The system was recovered and recycled with a 77% activity efficiency indicating recovery of the system and reusability. Preparation of a functional self-contained co-factor recycling system was demonstrated consisting of the biological components NADH oxidase and glucose dehydrogenase, immobilised on a polyethylenimine support with entrapped cofactor. This serves as proof-of-principle for the construction of derivative systems that could be used for the development of applications such as efficient biosynthesis, novel biosensors, diagnostic and therapeutic systems. 2014-07-30T17:42:12Z 2014-07-30T17:42:12Z 2010 Master Thesis Masters MSc http://hdl.handle.net/11427/4343 eng application/pdf Department of Molecular and Cell Biology Faculty of Science University of Cape Town |
| spellingShingle | Cell Biology Twala, Busisiwe V Preparation of a self-contained NADH co-factor recycling particle system |
| thesis_degree_str | Master's |
| title | Preparation of a self-contained NADH co-factor recycling particle system |
| title_full | Preparation of a self-contained NADH co-factor recycling particle system |
| title_fullStr | Preparation of a self-contained NADH co-factor recycling particle system |
| title_full_unstemmed | Preparation of a self-contained NADH co-factor recycling particle system |
| title_short | Preparation of a self-contained NADH co-factor recycling particle system |
| title_sort | preparation of a self contained nadh co factor recycling particle system |
| topic | Cell Biology |
| url | http://hdl.handle.net/11427/4343 |
| work_keys_str_mv | AT twalabusisiwev preparationofaselfcontainednadhcofactorrecyclingparticlesystem |