1. Low-cost anti-mycobacterial drug discovery using engineered E. coli.
- Author
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Bongaerts N, Edoo Z, Abukar AA, Song X, Sosa-Carrillo S, Haggenmueller S, Savigny J, Gontier S, Lindner AB, and Wintermute EH
- Subjects
- Antitubercular Agents chemistry, Antitubercular Agents pharmacology, Drug Discovery, Escherichia coli genetics, Escherichia coli metabolism, High-Throughput Screening Assays, Alanine Racemase chemistry, Alanine Racemase metabolism, Mycobacterium tuberculosis metabolism
- Abstract
Whole-cell screening for Mycobacterium tuberculosis (Mtb) inhibitors is complicated by the pathogen's slow growth and biocontainment requirements. Here we present a synthetic biology framework for assaying Mtb drug targets in engineered E. coli. We construct Target Essential Surrogate E. coli (TESEC) in which an essential metabolic enzyme is deleted and replaced with an Mtb-derived functional analog, linking bacterial growth to the activity of the target enzyme. High throughput screening of a TESEC model for Mtb alanine racemase (Alr) revealed benazepril as a targeted inhibitor, a result validated in whole-cell Mtb. In vitro biochemical assays indicated a noncompetitive mechanism unlike that of clinical Alr inhibitors. We establish the scalability of TESEC for drug discovery by characterizing TESEC strains for four additional targets., (© 2022. The Author(s).)
- Published
- 2022
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