Targeting DXP Synthase using TPP Mimics

Abstract

A global emergence of antibiotic‐resistant bacteria, largely due to the misuse and over use of antibiotics, has led to an increase in demand for the development of new antibacterial agents. In order to develop new antibiotics, researchers need to identify new targets that have high specificity for certain bacterial pathways. The enzyme D‐xylulose 5‐phosphate (DXP) synthase catalyzes the first step in the MEP (methylerythritolphosphate) pathway in human pathogens, which produces the biological molecules necessary to regulate numerous cellular functions, including central metabolism. Inhibition of DXP synthase would significantly reduce the rate of DXP production and the MEP pathway and thus is an ideal candidate for antibiotic development. DXP synthase requires the cofactor, thiamin pyrophosphate (TPP) for its activity. Preventing TPP from complexing with DXP synthase will inhibit the activity of the enzyme and consequently inhibit the MEP pathway. To test the efficacy of DXP synthase inhibitors both DXP synthase and the sequential enzyme of the MEP pathway, DXP reductoisomerase (IspC), were purified from E.coli using nickel affinity chromatography. Conversion from DXP to 2‐C‐Methyl‐D‐erythritol 4‐phosphate (MEP) by IspC coincides with NADPH oxidation that can be measured using spectroscopic techniques. The establishment of this in vitro system will allow us to test the efficacy novel DXP synthase inhibitors.Support or Funding InformationThis project was funded by SJFC internal funds.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.