Master of Science (MSc)
Biological and Chemical Sciences
Faculty of Science
Mentor during the research
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.
Torres Garcia, Eloy Jose, "Assessing the Biocatalytic Promiscuity of PEP- Transformations for Geminal-Difluorination" (2023). Theses and Dissertations (Comprehensive). 2540.
Available for download on Monday, January 26, 2026