Generation of Fluorinated Amychelin Siderophores against Pseudomonas aeruginosa Infections by a Combination of Genome Mining and Mutasynthesis

Feng Xie; Shengwang Dai; Yu Zhao; Pei Huang; Shen Yu; Biao Ren; Qiushui Wang; Zengchun Ji; Gil Alterovitz; Qi Zhang; Jingyu Zhang; Xiangyin Chen; Lan Jiang; Fuhang Song; Hongwei Liu; Frederick M Ausubel; Xueting Liu; Huanqin Dai; Lixin Zhang

doi:10.1016/j.chembiol.2020.10.009

Generation of Fluorinated Amychelin Siderophores against Pseudomonas aeruginosa Infections by a Combination of Genome Mining and Mutasynthesis

Cell Chem Biol. 2020 Dec 17;27(12):1532-1543.e6. doi: 10.1016/j.chembiol.2020.10.009. Epub 2020 Nov 12.

Authors

Feng Xie¹, Shengwang Dai², Yu Zhao², Pei Huang², Shen Yu³, Biao Ren⁴, Qiushui Wang², Zengchun Ji², Gil Alterovitz⁵, Qi Zhang⁶, Jingyu Zhang⁷, Xiangyin Chen⁷, Lan Jiang⁷, Fuhang Song², Hongwei Liu¹, Frederick M Ausubel³, Xueting Liu⁸, Huanqin Dai⁹, Lixin Zhang¹⁰

Affiliations

¹ State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
² Chinese Academy of Sciences Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
³ Department of Molecular Biology, Massachusetts General Hospital, Boston, MA 02114, USA.
⁴ State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases, Sichuan University, Chengdu, Sichuan 610041, China.
⁵ Boston Children's Hospital, Boston, MA 02115, USA.
⁶ Department of Pediatrics, Peking University China-Japan Friendship School of Clinical Medicine, Beijing 100029, China.
⁷ State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
⁸ State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China. Electronic address: liuxueting@ecust.edu.cn.
⁹ State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China. Electronic address: daihq@im.ac.cn.
¹⁰ State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China. Electronic address: lxzhang@ecust.edu.cn.

PMID: 33186541
DOI: 10.1016/j.chembiol.2020.10.009

Abstract

Pioneering microbial genomic surveys have revealed numerous untapped biosynthetic gene clusters, unveiling the great potential of new natural products. Here, using a combination of genome mining, mutasynthesis, and activity screening in an infection model comprising Caenorhabditis elegans and Pseudomonas aeruginosa, we identified candidate virulence-blocking amychelin siderophore compounds from actinomycetes. Subsequently, we developed unreported analogs of these virulence-blocking siderophores with improved potency by exploiting an Amycolatopsis methanolica strain 239^T chorismate to salicylate a biosynthetic subpathway for mutasynthesis. This allowed us to generate the fluorinated amychelin, fluoroamychelin I, which rescued C. elegans from P. aeruginosa-mediated killing with an EC₅₀ value of 1.4 μM, outperforming traditional antibiotics including ceftazidime and meropenem. In general, this paper describes an efficient platform for the identification and production of classes of anti-microbial compounds with potential unique modes of action.

Keywords: Caenorhabditis elegans; NRPS; Pseudomonas aeruginosa; actinobacteria; drug discovery; genome mining; host-pathogen interaction; mutasynthesis; pathogenesis; siderophore.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Anti-Bacterial Agents / chemistry
Anti-Bacterial Agents / pharmacology
Caenorhabditis elegans / genetics
Ceftazidime / pharmacology
Data Mining*
Drug Evaluation, Preclinical
Genomics*
Halogenation*
Meropenem / pharmacology
Pseudomonas aeruginosa / drug effects*
Pseudomonas aeruginosa / genetics*
Siderophores / chemistry*
Siderophores / pharmacology*

Substances

Anti-Bacterial Agents
Siderophores
Ceftazidime
Meropenem