The characterisation of the catalytic activity of human steroid 5α-reductase towards novel C19 substrates

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dc.contributor.advisorStorbeck, Karl-Heinzen_ZA
dc.contributor.authorQuanson, Jonathan Lukeen_ZA
dc.contributor.otherStellenbosch University. Faculty of Science. Dept. of Biochemistry.en_ZA
dc.date.accessioned2015-05-20T09:29:30Z
dc.date.available2015-05-20T09:29:30Z
dc.date.issued2015-02-25en
dc.date.issued2015-04
dc.descriptionThesis (MSc)--Stellenbosch University, 2015.en_ZA
dc.description.abstractENGLISH 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.en_ZA
dc.description.abstractAFRIKAANSE 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.af_ZA
dc.format.extentxi, 77 pages : illustrations (some colour)en_ZA
dc.identifier.urihttp://hdl.handle.net/10019.1/97057
dc.language.isoen_ZAen_ZA
dc.publisherStellenbosch : Stellenbosch Universityen_ZA
dc.rights.holderStellenbosch Universityen_ZA
dc.subjectProstate -- Cancer -- Treatmenten_ZA
dc.subjectSteroidogenesisen_ZA
dc.subjectPichia pastorisen_ZA
dc.subjectYeast expression systemen_ZA
dc.subjectUCTDen_ZA
dc.titleThe characterisation of the catalytic activity of human steroid 5α-reductase towards novel C19 substratesen_ZA
dc.typeThesisen_ZA
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