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

• Published in: Nature communications Vol. 13; no. 1; pp. 3905 - 11
• Main Authors: Bongaerts, Nadine, Edoo, Zainab, Abukar, Ayan A, Song, Xiaohu, Sosa-Carrillo, Sebastián, Haggenmueller, Sarah, Savigny, Juline, Gontier, Sophie, Lindner, Ariel B, Wintermute, Edwin H
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group 07.07.2022; Nature Publishing Group UK; Nature Portfolio
• Subjects: Alanine; Alanine racemase; Biology; Drug discovery; E coli; Enzymes; High-throughput screening; Inhibitors; Metabolism; Mycobacterium tuberculosis; Pathogens; Screening; Synthetic biology; Therapeutic targets; Tuberculosis
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-022-31570-3

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.