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Restricted Access Thesis

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First Advisor

Danielle Dube


Ralstonia is a genus that contains a variety of species that range from human pathogens to plant pathogens to bioremediators. Ralstonia can grow in water sources such as ponds, soil, laboratory-purified water systems, oxygen delivery systems, and dialysis water units. These common sources of contamination have led to nosocomial infections in immunocompromised patients. Due to certain Ralstonia species with innate resistance towards multiple β-lactam antibiotics, this raises concerns about future antibiotic resistance. The ineffectiveness of existing antibiotics supports the need to develop new approaches that interfere with the virulence of Ralstonia. Ralstonia glycoproteins and the enzymes responsible for their biosynthesis are intriguing targets because of their importance in fitness attributes such as growth, motility, and biofilm formation. In this work, we began by identifying a Ralstonia spp. isolate through 16S sequencing, biochemical tests, and polymerase chain reaction. After identifying our isolate to be Ralstonia pickettii, we employed metabolic oligosaccharide engineering (MOE) to provide biochemical evidence of a glycosylation system in R. pickettii. Then we further explored the effect of metabolic glycan inhibitors, 3F-DATDG-diNAc (3F-DAT), 3F-FucNAc (3F-Fuc), DATDG-diNAc-OBn (BnDAT), and FucNAc-OBn (BnFucNAc), on glycoprotein biosynthesis and fitness. Our preliminary results indicate that the inhibitors 3F-DAT and BnDAT induce glycosylation defects and alterations in growth and biofilm in R. pickettii. Altogether, these results suggest that metabolic inhibitors based on monosaccharides have the potential to disrupt glycosylation in a range of bacteria, including the human pathogen R. pickettii.


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