Synthesis of a GlmU Bisubstrate Inhibitor
Year: 2020
Type: dissertation
Abstract: In an attempt to combat antibiotic resistance, new drugs are constantly being developed. One method of killing bacteria is to inhibit the formation of the cell wall, which is vital to the survival of the cell. This project involves the chemoenzymatic synthesis of a bisubstrate inhibitor of M. tuberculosis GlmU, as well as the synthesis of glucosamine-1-phosphate (GlcN-1-P), a commercially expensive substrate of GlmU. These two compounds were synthesized by making acetamides followed by phosphorylation using the enzyme NahK. Acetyltransferase kinetics and inhibition were assessed by using a continuous ultraviolet absorbance assay involving an Ellman’s reagent derivative and Michaelis-Menten kinetics. The bisubstrate inhibitor (EL44) inhibits GlmU competitively with respect to acetyl-CoA (AcCoA) with a Ki of 9 ± 2 μM, which makes it the strongest inhibitor among published M. tuberculosis GlmU inhibitors. It also noncompetitively inhibits GlmU with respect to GlcN-1-P with a Ki of 51 ± 6 μM. Overall, the two target compounds were successfully synthesized and the inhibitor exhibited inhibitory activity against the formation of N-acetylglucosamine-1-phosphate.
Source: TSpace (University of Toronto)
Author: Ethan Lee
Language: English
Subfield: Organic Chemistry
Field: Chemistry
Domain: Physical Sciences
Open Access status: green
OpenAlex ID: W3186936852