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

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.

Identiferoai:union.ndltd.org:netd.ac.za/oai:union.ndltd.org:sun/oai:scholar.sun.ac.za:10019.1/97057
Date04 1900
CreatorsQuanson, Jonathan Luke
ContributorsStorbeck, Karl-Heinz, Stellenbosch University. Faculty of Science. Dept. of Biochemistry.
PublisherStellenbosch : Stellenbosch University
Source SetsSouth African National ETD Portal
Languageen_ZA
Detected LanguageEnglish
TypeThesis
Formatxi, 77 pages : illustrations (some colour)
RightsStellenbosch University

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