Development of an agar-plug cultivation system for bioactivity assays of actinomycete strain collections

Natural products are an important source of lead compounds for the development of drug substances. Actinomycetes have been valuable especially for the discovery of antibiotics. Increasing occurrence of antibiotic resistance among bacterial pathogens has revived the interest in actinomycete natural p...

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Autores principales:	Nico Ortlieb, Elke Klenk, Andreas Kulik, Timo Horst Johannes Niedermeyer
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/c8268515c67e4ebc8aaebb2e4b96183c
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spelling	oai:doaj.org-article:c8268515c67e4ebc8aaebb2e4b96183c2021-11-11T07:14:35ZDevelopment of an agar-plug cultivation system for bioactivity assays of actinomycete strain collections1932-6203https://doaj.org/article/c8268515c67e4ebc8aaebb2e4b96183c2021-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8570476/?tool=EBIhttps://doaj.org/toc/1932-6203Natural products are an important source of lead compounds for the development of drug substances. Actinomycetes have been valuable especially for the discovery of antibiotics. Increasing occurrence of antibiotic resistance among bacterial pathogens has revived the interest in actinomycete natural product research. Actinobacteria produce a different set of natural products when cultivated on solid growth media compared with submersed culture. Bioactivity assays involving solid media (e.g. agar-plug assays) require manual manipulation of the strains and agar plugs. This is less convenient for the screening of larger strain collections of several hundred or thousand strains. Thus, the aim of this study was to develop a 96-well microplate-based system suitable for the screening of actinomycete strain collections in agar-plug assays. We developed a medium-throughput cultivation and agar-plug assay workflow that allows the convenient inoculation of solid agar plugs with actinomycete spore suspensions from a strain collection, and the transfer of the agar plugs to petri dishes to conduct agar-plug bioactivity assays. The development steps as well as the challenges that were overcome during the development (e.g. system sterility, handling of the agar plugs) are described. We present the results from one exemplary screening campaign targeted to identify compounds inhibiting Agr-based quorum sensing where the workflow was used successfully. We present a novel and convenient workflow to combine agar diffusion assays with microtiter-plate-based cultivation systems in which strains can grow on a solid surface. This workflow facilitates and speeds up the initial medium throughput screening of natural product-producing actinomycete strain collections against monitor strains in agar-plug assays.Nico OrtliebElke KlenkAndreas KulikTimo Horst Johannes NiedermeyerPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Nico Ortlieb Elke Klenk Andreas Kulik Timo Horst Johannes Niedermeyer Development of an agar-plug cultivation system for bioactivity assays of actinomycete strain collections
description	Natural products are an important source of lead compounds for the development of drug substances. Actinomycetes have been valuable especially for the discovery of antibiotics. Increasing occurrence of antibiotic resistance among bacterial pathogens has revived the interest in actinomycete natural product research. Actinobacteria produce a different set of natural products when cultivated on solid growth media compared with submersed culture. Bioactivity assays involving solid media (e.g. agar-plug assays) require manual manipulation of the strains and agar plugs. This is less convenient for the screening of larger strain collections of several hundred or thousand strains. Thus, the aim of this study was to develop a 96-well microplate-based system suitable for the screening of actinomycete strain collections in agar-plug assays. We developed a medium-throughput cultivation and agar-plug assay workflow that allows the convenient inoculation of solid agar plugs with actinomycete spore suspensions from a strain collection, and the transfer of the agar plugs to petri dishes to conduct agar-plug bioactivity assays. The development steps as well as the challenges that were overcome during the development (e.g. system sterility, handling of the agar plugs) are described. We present the results from one exemplary screening campaign targeted to identify compounds inhibiting Agr-based quorum sensing where the workflow was used successfully. We present a novel and convenient workflow to combine agar diffusion assays with microtiter-plate-based cultivation systems in which strains can grow on a solid surface. This workflow facilitates and speeds up the initial medium throughput screening of natural product-producing actinomycete strain collections against monitor strains in agar-plug assays.
format	article
author	Nico Ortlieb Elke Klenk Andreas Kulik Timo Horst Johannes Niedermeyer
author_facet	Nico Ortlieb Elke Klenk Andreas Kulik Timo Horst Johannes Niedermeyer
author_sort	Nico Ortlieb
title	Development of an agar-plug cultivation system for bioactivity assays of actinomycete strain collections
title_short	Development of an agar-plug cultivation system for bioactivity assays of actinomycete strain collections
title_full	Development of an agar-plug cultivation system for bioactivity assays of actinomycete strain collections
title_fullStr	Development of an agar-plug cultivation system for bioactivity assays of actinomycete strain collections
title_full_unstemmed	Development of an agar-plug cultivation system for bioactivity assays of actinomycete strain collections
title_sort	development of an agar-plug cultivation system for bioactivity assays of actinomycete strain collections
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/c8268515c67e4ebc8aaebb2e4b96183c
work_keys_str_mv	AT nicoortlieb developmentofanagarplugcultivationsystemforbioactivityassaysofactinomycetestraincollections AT elkeklenk developmentofanagarplugcultivationsystemforbioactivityassaysofactinomycetestraincollections AT andreaskulik developmentofanagarplugcultivationsystemforbioactivityassaysofactinomycetestraincollections AT timohorstjohannesniedermeyer developmentofanagarplugcultivationsystemforbioactivityassaysofactinomycetestraincollections
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Development of an agar-plug cultivation system for bioactivity assays of actinomycete strain collections

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