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The characterisation of the catalytic activity of human steroid 5α-reductase towards novel C19 substrates
Thesis (MSc)--Stellenbosch University, 2015.
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| Format: | Thesis |
| Language: | en_ZA |
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Stellenbosch : Stellenbosch University
2015
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| _version_ | 1867613951472173056 |
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| access_status_str | Open Access |
| author | Quanson, Jonathan Luke |
| author2 | Storbeck, Karl-Heinz |
| author_browse | Quanson, Jonathan Luke Storbeck, Karl-Heinz |
| author_facet | Storbeck, Karl-Heinz Quanson, Jonathan Luke |
| author_sort | Quanson, Jonathan Luke |
| collection | Thesis |
| dc_rights_str_mv | Stellenbosch University |
| description | Thesis (MSc)--Stellenbosch University, 2015. |
| format | Thesis |
| id | oai:scholar.sun.ac.za:10019.1/97057 |
| institution | Stellenbosch University (South Africa) |
| language | en_ZA |
| last_indexed | 2026-06-10T12:44:18.274Z |
| license_str | Other — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from SUNScholar — Stellenbosch University Repository |
| publishDate | 2015 |
| publishDateRange | 2015 |
| publishDateSort | 2015 |
| publisher | Stellenbosch : Stellenbosch University |
| publisherStr | Stellenbosch : Stellenbosch University |
| record_format | dspace |
| source_str | SUNScholar — Stellenbosch University Repository |
| spelling | oai:scholar.sun.ac.za:10019.1/97057 The characterisation of the catalytic activity of human steroid 5α-reductase towards novel C19 substrates Quanson, Jonathan Luke Storbeck, Karl-Heinz Stellenbosch University. Faculty of Science. Dept. of Biochemistry. Prostate -- Cancer -- Treatment Steroidogenesis Pichia pastoris Yeast expression system UCTD Thesis (MSc)--Stellenbosch University, 2015. ENGLISH ABSTRACT: This study describes: • The UPLC-MS/MS analyses and quantification of novel 5α-reduced steroids using response factors. • The kinetic characterisation of human steroid 5α-reductase type 1 (SRD5A1), expressed in HEK-293 cells, towards 11OHA4 and 11OHT and their keto derivatives by progress curve analysis. • The subcloning, transformation and functional expression of SRD5A1 in the yeast expression system, P. pastoris. • The conversion of 11OHA4 and 11OHT and their keto derivatives by SRD5A1 expressed in P. pastoris. • The endogenous enzymatic activity in P. pastoris towards the 5α-reduced metabolites in the 11OHA4- and alternate 5α-dione pathways. • The potential application of P. pastoris as a biocatalyst in the production of 5α- reduced C19 steroids. AFRIKAANSE OPSOMMING: Hierdie ondersoek beskryf: • Die UPLC-MS/MS analise en kwantifisering van nuut-ondekte 5α-gereduseerde steroïede met behulp van responsfaktore. • Die kinetiese karakterisering van menslike steroïed 5α-reduktase tipe 1 (SRD5A1), uitgedruk in HEK-293 selle, vir 11OHA4 en 11OHT en hul ketoderivate deur middel van progressiekurwe-analise. • Die subklonering, transformasie en funksionele uitdrukking van SRD5A1 in die gis P. pastoris. • Die omsetting van 11OHA4 en 11OHT en hul ketoderivate deur SRD5A1 uitgedruk in P. pastoris. • Die omsetting van 5α-gereduseerde steroïede in die 11OHA4 en alternatiewe 5α-dioon paaie deur endogene ensieme in P. pastoris • ‘n Ondersoek na die toepassing van die gisuitdrukkingstelsel as ‘n moontlike OR potensiële biokatalis vir die produksie van 5α-gereduseerde C19 steroïede. 2015-05-20T09:29:30Z 2015-05-20T09:29:30Z 2015-02-25 2015-04 Thesis http://hdl.handle.net/10019.1/97057 en_ZA Stellenbosch University xi, 77 pages : illustrations (some colour) application/pdf Stellenbosch : Stellenbosch University |
| spellingShingle | Prostate -- Cancer -- Treatment Steroidogenesis Pichia pastoris Yeast expression system UCTD Quanson, Jonathan Luke The characterisation of the catalytic activity of human steroid 5α-reductase towards novel C19 substrates |
| title | The characterisation of the catalytic activity of human steroid 5α-reductase towards novel C19 substrates |
| title_full | The characterisation of the catalytic activity of human steroid 5α-reductase towards novel C19 substrates |
| title_fullStr | The characterisation of the catalytic activity of human steroid 5α-reductase towards novel C19 substrates |
| title_full_unstemmed | The characterisation of the catalytic activity of human steroid 5α-reductase towards novel C19 substrates |
| title_short | The characterisation of the catalytic activity of human steroid 5α-reductase towards novel C19 substrates |
| title_sort | characterisation of the catalytic activity of human steroid 5α reductase towards novel c19 substrates |
| topic | Prostate -- Cancer -- Treatment Steroidogenesis Pichia pastoris Yeast expression system UCTD |
| url | http://hdl.handle.net/10019.1/97057 |
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