Review the symposium abstracts (3.19MB PDF) - College of Science

Poster PresentationSynthesis of a Hydroxybenzoate Ligand for a Class 1A Dihydroorotate DehydrogenaseChristine CuthbertsonAdvisor: Bruce Palfey, Dept. of Biological Chemistry, University of MichiganDihydroorotate dehydrogenases (DHODs) catalyze the only redox reaction in pyrimidinebiosynthesis—the oxidation of dihydroorotate to orotate. Class 1A DHODs are found in somedisease-causing protozoans, which make them possible drug targets in the treatment ofLeishmaniasis, Chagas’ disease, and African sleeping sickness. An enzymatic route forsynthesizing ligands has been established utilizing the enzyme p-hydroxybenzoate hydroxylase(PHBH). PHBH was overexpressed using the plasmid pIE-130 in Escherichia coli strain JM105and purified by salting-out some impurities with ammonium sulfate, dialyzing, treating withheat, followed by affinity chromatography using a Red-Dye column. A ligand was made by thehydroxylation of hydroxybenzoate-precursor by PHBH. Purification was completed by HPLCand the ligand was verified by UV-vis spectrophotometry and NMR spectroscopy.43