A high-throughput, whole cell assay to identify compounds active against carbapenem-resistant Klebsiella pneumoniae.

Enteric Gram-negative rods (GNR), which are frequent causes of community-acquired and nosocomial infections, are increasingly resistant to the antibiotics in our current armamentarium. One solution to this medical dilemma is the development of novel classes of antimicrobial compounds. Here we report...

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Main Authors: Julie Liao, George Xu, Emily E Mevers, Jon Clardy, Paula I Watnick
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2018-01-01
Series:PLoS ONE
Online Access:https://doi.org/10.1371/journal.pone.0209389
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author Julie Liao
George Xu
Emily E Mevers
Jon Clardy
Paula I Watnick
author_facet Julie Liao
George Xu
Emily E Mevers
Jon Clardy
Paula I Watnick
author_sort Julie Liao
collection DOAJ
description Enteric Gram-negative rods (GNR), which are frequent causes of community-acquired and nosocomial infections, are increasingly resistant to the antibiotics in our current armamentarium. One solution to this medical dilemma is the development of novel classes of antimicrobial compounds. Here we report the development of a robust, whole cell-based, high-throughput metabolic assay that detects compounds with activity against carbapenem-resistant Klebsiella pneumoniae. We have used this assay to screen approximately 8,000 fungal extracts and 50,000 synthetic compounds with the goal of identifying extracts and compounds active against a highly resistant strain of Klebsiella pneumoniae. The primary screen identified 43 active fungal extracts and 144 active synthetic compounds. Patulin, a known fungal metabolite and inhibitor of bacterial quorum sensing and alanine racemase, was identified as the active component in the most potent fungal extracts. We did not study patulin further due to previously published evidence of toxicity. Three synthetic compounds termed O06, C17, and N08 were chosen for further study. Compound O06 did not have significant antibacterial activity but rather interfered with sugar metabolism, while compound C17 had only moderate activity against GNRs. Compound N08 was active against several resistant GNRs and showed minimal toxicity to mammalian cells. Preliminary studies suggested that it interferes with protein expression. However, its direct application may be limited by susceptibility to efflux and a tendency to form aggregates in aqueous media. Rapid screening of 58,000 test samples with identification of several compounds that act on CR-K. pneumoniae demonstrates the utility of this screen for the discovery of drugs active against this highly resistant GNR.
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spelling doaj.art-749e202bb89d4b46935d46e192afb0032022-12-21T18:38:48ZengPublic Library of Science (PLoS)PLoS ONE1932-62032018-01-011312e020938910.1371/journal.pone.0209389A high-throughput, whole cell assay to identify compounds active against carbapenem-resistant Klebsiella pneumoniae.Julie LiaoGeorge XuEmily E MeversJon ClardyPaula I WatnickEnteric Gram-negative rods (GNR), which are frequent causes of community-acquired and nosocomial infections, are increasingly resistant to the antibiotics in our current armamentarium. One solution to this medical dilemma is the development of novel classes of antimicrobial compounds. Here we report the development of a robust, whole cell-based, high-throughput metabolic assay that detects compounds with activity against carbapenem-resistant Klebsiella pneumoniae. We have used this assay to screen approximately 8,000 fungal extracts and 50,000 synthetic compounds with the goal of identifying extracts and compounds active against a highly resistant strain of Klebsiella pneumoniae. The primary screen identified 43 active fungal extracts and 144 active synthetic compounds. Patulin, a known fungal metabolite and inhibitor of bacterial quorum sensing and alanine racemase, was identified as the active component in the most potent fungal extracts. We did not study patulin further due to previously published evidence of toxicity. Three synthetic compounds termed O06, C17, and N08 were chosen for further study. Compound O06 did not have significant antibacterial activity but rather interfered with sugar metabolism, while compound C17 had only moderate activity against GNRs. Compound N08 was active against several resistant GNRs and showed minimal toxicity to mammalian cells. Preliminary studies suggested that it interferes with protein expression. However, its direct application may be limited by susceptibility to efflux and a tendency to form aggregates in aqueous media. Rapid screening of 58,000 test samples with identification of several compounds that act on CR-K. pneumoniae demonstrates the utility of this screen for the discovery of drugs active against this highly resistant GNR.https://doi.org/10.1371/journal.pone.0209389
spellingShingle Julie Liao
George Xu
Emily E Mevers
Jon Clardy
Paula I Watnick
A high-throughput, whole cell assay to identify compounds active against carbapenem-resistant Klebsiella pneumoniae.
PLoS ONE
title A high-throughput, whole cell assay to identify compounds active against carbapenem-resistant Klebsiella pneumoniae.
title_full A high-throughput, whole cell assay to identify compounds active against carbapenem-resistant Klebsiella pneumoniae.
title_fullStr A high-throughput, whole cell assay to identify compounds active against carbapenem-resistant Klebsiella pneumoniae.
title_full_unstemmed A high-throughput, whole cell assay to identify compounds active against carbapenem-resistant Klebsiella pneumoniae.
title_short A high-throughput, whole cell assay to identify compounds active against carbapenem-resistant Klebsiella pneumoniae.
title_sort high throughput whole cell assay to identify compounds active against carbapenem resistant klebsiella pneumoniae
url https://doi.org/10.1371/journal.pone.0209389
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