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Utilization of Vibrio cholerae as a Model Organism to Screen Natural Product Libraries for Identification of New Antibiotics

  • Aleksandra E. SikoraEmail author
  • Richard Tehan
  • Kerry McPhail
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1839)

Abstract

The development of antibiotic-resistant bacteria requires increasing research efforts in drug discovery. Vibrio cholerae can be utilized as a model gram-negative enteric pathogen in high- and medium-throughput screening campaigns to identify antimicrobials with different modes of action. In this chapter, we describe methods for the optimal growth of V. cholerae in 384-well plates, preparation of suitable microtiter natural product sample libraries, as well as their screening using measurements of bacterial density and activity of type II secretion-dependent protease as readouts. Concomitant LC-MS/MS profiling and spectral data networking of assay sample libraries facilitate dereplication of putative known and/or nuisance compounds and efficient prioritization of samples containing putative new natural products for further investigation.

Key words

Type II secretion system Serine proteases Cell-based assay Natural products High-throughput screening LC-MS 

Notes

Acknowledgments

This work was funded by the start-up funds to A.E.S. and an Oregon Sea Grant under award number NA10OAR4170059 (project number R/BT-48) from the National Oceanic and Atmospheric Administration’s National Sea Grant College Program to K.L.M. and A.E.S.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Aleksandra E. Sikora
    • 1
    Email author
  • Richard Tehan
    • 1
  • Kerry McPhail
    • 1
  1. 1.Department of Pharmaceutical Sciences, College of PharmacyOregon State UniversityCorvallisUSA

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