Discovery of External Modulators of the Fe-Fe Hydrogenase Enzyme in Clostridium acetobutylicum

Report No. ARL-TR-7189
Authors: Michael S Lee; Margaret M Hurley; Matthew Servinsky; Christian Sund
Date/Pages: February 2015; 30 pages
Abstract: Complex metabolic networks contain many essential enzymes that cannot be genetically altered or deleted without adverse impact on the host cell. We have developed an alternate strategy to modulate metabolite flow through these essential enzymes with the goal of more efficient production of commodity chemicals (i.e., biofuels), better waste remediation, and novel antibiotics. In this report we tackle the first of 2 goals to computationally discover and design small molecules and modified bacterial genes (for producing peptide tails to a common bacterial protein) that will inhibit native bacterial metabolic enzymes and alter metabolic output. We computationally screened the million-compound ChemDiv catalogue and identified and purchased 20 candidate inhibitors to 2 sites of the Fe-Fe hydrogenase protein in Clostridium acetobutylicum. During this effort, we developed a novel assay for detecting gas production in ultrasmall bacterial cultures and verified this assay with TNT as a positive control. After testing all of the candidate molecules in this assay, we found some that moderately reduced hydrogen gas production as desired and some that reduced overall cell viability.
Distribution: Approved for public release
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Last Update / Reviewed: February 1, 2015