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Karakterisering van die katalase van BCG : invloed van isoniasied op die gesuiwerde ensiem

Thesis (MSc)--Stellenbosch University, 1990.

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Main Author: Basson, Karen
Other Authors: Van Zyl, J. M.
Format: Thesis
Language:af_ZA
Published: Stellenbosch : Stellenbosch University 2012
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access_status_str Open Access
author Basson, Karen
author2 Van Zyl, J. M.
author_browse Basson, Karen
Van Zyl, J. M.
author_facet Van Zyl, J. M.
Basson, Karen
author_sort Basson, Karen
collection Thesis
dc_rights_str_mv Stellenbosch University
description Thesis (MSc)--Stellenbosch University, 1990.
format Thesis
id oai:scholar.sun.ac.za:10019.1/68852
institution Stellenbosch University (South Africa)
language af_ZA
last_indexed 2026-06-10T12:46:21.556Z
license_str Other — see source repository
provenance_str_mv Harvested via OAI-PMH from SUNScholar — Stellenbosch University Repository
publishDate 2012
publishDateRange 2012
publishDateSort 2012
publisher Stellenbosch : Stellenbosch University
publisherStr Stellenbosch : Stellenbosch University
record_format dspace
source_str SUNScholar — Stellenbosch University Repository
spelling oai:scholar.sun.ac.za:10019.1/68852 Karakterisering van die katalase van BCG : invloed van isoniasied op die gesuiwerde ensiem Basson, Karen Van Zyl, J. M. Van der Walt, B. J. Stellenbosch University. Faculty of Medicine and Health Sciences. Dept. of Clinical Pharmacology. BCG vaccines Tuberculosis vaccines Chromatography Hydroxyapatite Catalase Thesis (MSc)--Stellenbosch University, 1990. ENGLISH ABSTRACT: Catalase was extracted and isolated from BCG bacilli by the following precedures : sonication, treatment with deoxyribonuclease, treatment with octyl glucoside and sonification, ammonium sulphate precipitation, hydroxyapatite chromatography and chromatography on Sephacryl S-1000. A significant increase in the specific activity of the catalase and peroxidase functions was observed during purification and a final purity index (A405/A280) of 0.258 was obtained. Only one protein band, in addition to aggregates which could hardly penetrate the gel, was observed after non-denaturing polyacrylamide gel electrophoresis of the purified enzyme. The peroxidase activity (ABTS) assay) was consistent with the protein band. A.M. of 163.1 kDa was calculated from results obtained with SDS polyacrylamide gel electrophoresis, which correlates with a Mr of 144.54 kDa obtained by gelfiltration studies on Sephacryl S-1000. A characteristic Soret peak at 405 nm was observed on the UV spectrum of the purified enzyme. The optimum pH for catalase activity was 8,0. At pH 7,0 81% of the maximum activity at pH 6,0 and 56% at pH 9,0. This correlates with the fact that catalase must be able to survive over a broad pH range. AFRIKAANSE OPSOMING: Katalase is geëkstraheer uit BCG basille en geïsoleer deur die volgende opeenvolgende stappe: sonikering, behandeling met deoksiribonuklease, behandeling met oktielglukosied en sonikering, ammoniumsulfaat presipitasie, hidroksi-apatiet chromatografie en chromatografie op Sephacryl S-1000. Die spesiefieke aktiwiteit van die katalatiese en peroksidatiewe funksies het beduidend toegeneem tydens suiwering en 'n finale RZ-waarde (A405/A280) van 0,258 is verkry. Analitiese poli-akrielanied gelelektroforese van die gesuiwerde ensien het hoofsaaklik een band getoon. Daar was ooreenstemming tussen die proteïenband en peroksidase aktiwiteit (ABTS bepaling). 'n Mr van 163,1 kDa is mbv SDS-PAGE bereken, wat ooreenstem met 'n Mr van 144,54 kDa wat bepaal is mbv gelfiltrasie studies op 'n Sephacryl S-1000 kolon. Die UV spektrum van die gesuiwerde ensiem het 'n kenmerkende Soret piek by 405nm getoon. Die optimum pH vir katalase aktiwiteit was 8,0. By neutrale pH was die aktiwiteit nog 81% van die maksimum waarde. By pH 6,0 het die ensiem nog 49% van die maksimum aktiwiteit getoon, terwyl 'n waarde van 56% by pH 9,0 gevind is. Hierdie bevindings is in ooreenstemming daarmee dat katalase oor 'n relatiewe breë pH gebied moet kan oorleef. Masters 2012-08-27T12:26:48Z 2012-08-27T12:26:48Z 1990-12 Thesis http://hdl.handle.net/10019.1/68852 af_ZA Stellenbosch University 148 pages application/pdf Stellenbosch : Stellenbosch University
spellingShingle BCG vaccines
Tuberculosis vaccines
Chromatography
Hydroxyapatite
Catalase
Basson, Karen
Karakterisering van die katalase van BCG : invloed van isoniasied op die gesuiwerde ensiem
title Karakterisering van die katalase van BCG : invloed van isoniasied op die gesuiwerde ensiem
title_full Karakterisering van die katalase van BCG : invloed van isoniasied op die gesuiwerde ensiem
title_fullStr Karakterisering van die katalase van BCG : invloed van isoniasied op die gesuiwerde ensiem
title_full_unstemmed Karakterisering van die katalase van BCG : invloed van isoniasied op die gesuiwerde ensiem
title_short Karakterisering van die katalase van BCG : invloed van isoniasied op die gesuiwerde ensiem
title_sort karakterisering van die katalase van bcg invloed van isoniasied op die gesuiwerde ensiem
topic BCG vaccines
Tuberculosis vaccines
Chromatography
Hydroxyapatite
Catalase
url http://hdl.handle.net/10019.1/68852
work_keys_str_mv AT bassonkaren karakteriseringvandiekatalasevanbcginvloedvanisoniasiedopdiegesuiwerdeensiem