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Low-cost anti-mycobacterial drug discovery using engineered E. coli.

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.

SUBMITTER: Bongaerts N

PROVIDER: S-EPMC9262897 | biostudies-literature | 2022 Jul

REPOSITORIES: biostudies-literature

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Low-cost anti-mycobacterial drug discovery using engineered E. coli.

Bongaerts Nadine N Edoo Zainab Z Abukar Ayan A AA Song Xiaohu X Sosa-Carrillo Sebastián S Haggenmueller Sarah S Savigny Juline J Gontier Sophie S Lindner Ariel B AB Wintermute Edwin H EH

Nature communications 20220707 1

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 TE ...[more]

PMID: 35798732

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