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Darobactins Exhibiting Superior Antibiotic Activity by Cryo-EM Structure Guided Biosynthetic Engineering** |
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| Authors: | Carsten E Seyfert Christoph Porten Dr Biao Yuan Selina Deckarm Dr Fabian Panter Dr Chantal D Bader Janetta Coetzee Felix Deschner Dr Kamaleddin H M E Tehrani Prof Dr Paul G Higgins Prof Dr Harald Seifert Prof Dr Thomas C Marlovits Dr Jennifer Herrmann Prof Dr Rolf Müller |
| Institution: | 1. Microbial Natural Products, Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Centre for Infection Research (HZI), Department of Pharmacy at, Saarland University Campus Building E8.1, 66123 Saarbrücken (Germany);2. University Medical Center Hamburg-Eppendorf (UKE), Institute of Structural and Systems Biology, Notkestraße 85, Building 15, 22607 Hamburg, Germany
Centre for Structural Systems Biology (CSSB), Hamburg, Germany These authors contributed equally to this work.;3. Microbial Natural Products, Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Centre for Infection Research (HZI), Department of Pharmacy at, Saarland University Campus Building E8.1, 66123 Saarbrücken (Germany) German Centre for Infection Research (DZIF), partnersite Hannover-Braunschweig, Germany;4. Microbiology, Immunology and Hygiene, Faculty of Medicine and University Hospital Cologne, University of Cologne, Cologne, Germany;5. University Medical Center Hamburg-Eppendorf (UKE), Institute of Structural and Systems Biology, Notkestraße 85, Building 15, 22607 Hamburg, Germany |
| Abstract: | Over recent decades, the pipeline of antibiotics acting against Gram-negative bacteria is running dry, as most discovered candidate antibiotics suffer from insufficient potency, pharmacokinetic properties, or toxicity. The darobactins, a promising new small peptide class of drug candidates, bind to novel antibiotic target BamA, an outer membrane protein. Previously, we reported that biosynthetic engineering in a heterologous host generated novel darobactins with enhanced antibacterial activity. Here we utilize an optimized purification method and present cryo-EM structures of the Bam complex with darobactin 9 (D9), which served as a blueprint for the biotechnological generation of twenty new darobactins including halogenated analogs. The newly engineered darobactin 22 binds more tightly to BamA and outperforms the favorable activity profile of D9 against clinically relevant pathogens such as carbapenem-resistant Acinetobacter baumannii up to 32-fold, without observing toxic effects. |
| Keywords: | Antibiotics BamA CRAB Cryo-Electron Microscopy Natural Products |