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Functional and structural characterisation of a multimodular xylanase from the hindgut metagenome of the snouted harvester termite
Dissertation (MSc)--University of Pretoria, 2017.
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| Format: | Thesis |
| Language: | English |
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University of Pretoria
2018
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| _version_ | 1867613472766820352 |
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| access_status_str | Open Access |
| author2 | Schubert, Wolf-Dieter |
| author_browse | Schubert, Wolf-Dieter |
| author_facet | Schubert, Wolf-Dieter |
| collection | Thesis |
| dc_rights_str_mv | © 2018 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. |
| description | Dissertation (MSc)--University of Pretoria, 2017. |
| format | Thesis |
| id | oai:repository.up.ac.za:2263/65901 |
| institution | University of Pretoria (South Africa) |
| language | English |
| last_indexed | 2026-06-10T12:36:41.872Z |
| license_str | Other — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from UPSpace — University of Pretoria Institutional Repository |
| publishDate | 2018 |
| publishDateRange | 2018 |
| publishDateSort | 2018 |
| publisher | University of Pretoria |
| publisherStr | University of Pretoria |
| record_format | dspace |
| source_str | UPSpace — University of Pretoria Institutional Repository |
| spelling | oai:repository.up.ac.za:2263/65901 Functional and structural characterisation of a multimodular xylanase from the hindgut metagenome of the snouted harvester termite Schubert, Wolf-Dieter robert_kru@yahoo.com Motshwene, Precious G. Kruger, Robert Friedrich UCTD Dissertation (MSc)--University of Pretoria, 2017. The degradation of hemicellulose is a vital step in the efficient utilisation of plant biomass, an abundant source of carbohydrates in nature. Xylan, the most prevalent hemicellulose, is digested by xylanases and much work has gone into improving the efficiency and stability of these enzymes. Interest has recently shifted to non-catalytic, accessory domains found in many xylanases, including carbohydrate binding modules (CBMs). CBMs may enhance enzyme properties, such as thermostability and efficiency. This effect has, however, not yet been fully explained. In this study, a novel multimodular xylanase isolated from the hindgut metagenome of the snouted harvester termite (Trinervitermes trinervoides) was characterised and the properties of its two identical CBMs analysed. Xylanase variants were produced in E. coli by deleting none, one or both CBMs. The variants were functionally characterised using a modified DNS assay in a thermocycler and structurally modelled using computational techniques. Deleting one CBM shifted the pH profile, slightly increasing activity at lower pH whereas removing both CBMs decreased activity at 60°C from 90% (SD = 1.3) to 56% (SD = 2.9). Removal of the CBMs did not statistically significantly affect the Michaelis constants (KM), but did reduce the turnover number (kcat). Structure modelling failed to accurately predict the structure of the complete enzyme and in particular any interactions between domains. The origin of the stabilising effect of the second CBM and hence the evolutionary benefit of two CBMs remains unresolved. Biochemistry MSc Unrestricted 2018-07-25T09:00:49Z 2018-07-25T09:00:49Z 2018/04/24 2017 Dissertation Kruger, RF 2017, Functional and structural characterisation of a multimodular xylanase from the hindgut metagenome of the snouted harvester termite, MSc Dissertation, University of Pretoria, Pretoria, viewed yymmdd <http://hdl.handle.net/2263/65901> A2018 http://hdl.handle.net/2263/65901 en © 2018 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. application/pdf University of Pretoria |
| spellingShingle | UCTD Functional and structural characterisation of a multimodular xylanase from the hindgut metagenome of the snouted harvester termite |
| title | Functional and structural characterisation of a multimodular xylanase from the hindgut metagenome of the snouted harvester termite |
| title_full | Functional and structural characterisation of a multimodular xylanase from the hindgut metagenome of the snouted harvester termite |
| title_fullStr | Functional and structural characterisation of a multimodular xylanase from the hindgut metagenome of the snouted harvester termite |
| title_full_unstemmed | Functional and structural characterisation of a multimodular xylanase from the hindgut metagenome of the snouted harvester termite |
| title_short | Functional and structural characterisation of a multimodular xylanase from the hindgut metagenome of the snouted harvester termite |
| title_sort | functional and structural characterisation of a multimodular xylanase from the hindgut metagenome of the snouted harvester termite |
| topic | UCTD |
| url | http://hdl.handle.net/2263/65901 |