Document Type

Thesis

Degree Name

Master of Science (MSc)

Department

Chemistry

Program Name/Specialization

Biological and Chemical Sciences

Faculty/School

Faculty of Science

First Advisor

Geoff Horsman

Advisor Role

Mentor during the research

Abstract

Fluorine is the most electronegative chemical element and can drastically change molecular properties, such as drug pharmacokinetics and agrochemical half-life. Although a great deal of progress is occurring in synthetic methods to access various organofluorines, these methods struggle with harsh conditions and limited substrate scope. Biocatalytic approaches offer the potential to access organofluorines that are difficult or impossible with the current methodology. Phosphoenolpyruvate (PEP) is a central metabolite and biosynthetic precursor for diverse molecules. Phosphoenol pyruvate mutase (Ppm) catalyses the interconversion of Phosphonopyruvate (PnPy) to Phosphoenolpyruvate (PEP). This research successfully expressed and purified three recombinant enzymes: Phosphosulfolactate synthase (PLS), Sialic acid synthase (NeuB) and Olsenella uli – Ppm (Oul-Ppm). This work also developed a HILIC method accompanied by ESI-HR MS/MS, which determined the molecular fingerprint of DF-PnPy and DF-PEP catalysed by a recombinant Ppm (Oul-Ppm). DF-PEP was coupled with a recombinant PLS that catalysed the addition of HSO3 to DF-PEP to produce a new organofluorine molecule, Difluoro-Phophosulfolactone [DF-PEP-SO3]; this molecule was analysed using a novel HILIC/ESI-HR MS/MS method acquiring its molecular fingerprint. In conclusion, this research expressed and purified PLS, NeuB and Oul-Ppm and established the first steps toward a biocatalytic platform using a difluorinated PEP to produce gem-difluoro-containing natural products.

Convocation Year

2023

Convocation Season

Spring

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Available for download on Monday, January 26, 2026

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