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Genetic Engineering of Streptomyces ghanaensis ATCC14672 for Improved Production of Moenomycins

Streptomycetes are soil-dwelling multicellular microorganisms famous for their unprecedented ability to synthesize numerous bioactive natural products (NPs). In addition to their rich arsenal of secondary metabolites, Streptomyces are characterized by complex morphological differentiation. Mostly, i...

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Autores principales: Makitrynskyy, Roman, Tsypik, Olga, Bechthold, Andreas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8778134/
https://www.ncbi.nlm.nih.gov/pubmed/35056478
http://dx.doi.org/10.3390/microorganisms10010030
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author Makitrynskyy, Roman
Tsypik, Olga
Bechthold, Andreas
author_facet Makitrynskyy, Roman
Tsypik, Olga
Bechthold, Andreas
author_sort Makitrynskyy, Roman
collection PubMed
description Streptomycetes are soil-dwelling multicellular microorganisms famous for their unprecedented ability to synthesize numerous bioactive natural products (NPs). In addition to their rich arsenal of secondary metabolites, Streptomyces are characterized by complex morphological differentiation. Mostly, industrial production of NPs is done by submerged fermentation, where streptomycetes grow as a vegetative mycelium forming pellets. Often, suboptimal growth peculiarities are the major bottleneck for industrial exploitation. In this work, we employed genetic engineering approaches to improve the production of moenomycins (Mm) in Streptomyces ghanaensis, the only known natural direct inhibitors of bacterial peptidoglycan glycosyltransferses. We showed that in vivo elimination of binding sites for the pleiotropic regulator AdpA in the oriC region strongly influences growth and positively correlates with Mm accumulation. Additionally, a marker- and “scar”-less deletion of moeH5, encoding an amidotransferase from the Mm gene cluster, significantly narrows down the Mm production spectrum. Strikingly, antibiotic titers were strongly enhanced by the elimination of the pleiotropic regulatory gene wblA, involved in the late steps of morphogenesis. Altogether, we generated Mm overproducers with optimized growth parameters, which are useful for further genome engineering and chemoenzymatic generation of novel Mm derivatives. Analogously, such a scheme can be applied to other Streptomyces spp.
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spelling pubmed-87781342022-01-22 Genetic Engineering of Streptomyces ghanaensis ATCC14672 for Improved Production of Moenomycins Makitrynskyy, Roman Tsypik, Olga Bechthold, Andreas Microorganisms Article Streptomycetes are soil-dwelling multicellular microorganisms famous for their unprecedented ability to synthesize numerous bioactive natural products (NPs). In addition to their rich arsenal of secondary metabolites, Streptomyces are characterized by complex morphological differentiation. Mostly, industrial production of NPs is done by submerged fermentation, where streptomycetes grow as a vegetative mycelium forming pellets. Often, suboptimal growth peculiarities are the major bottleneck for industrial exploitation. In this work, we employed genetic engineering approaches to improve the production of moenomycins (Mm) in Streptomyces ghanaensis, the only known natural direct inhibitors of bacterial peptidoglycan glycosyltransferses. We showed that in vivo elimination of binding sites for the pleiotropic regulator AdpA in the oriC region strongly influences growth and positively correlates with Mm accumulation. Additionally, a marker- and “scar”-less deletion of moeH5, encoding an amidotransferase from the Mm gene cluster, significantly narrows down the Mm production spectrum. Strikingly, antibiotic titers were strongly enhanced by the elimination of the pleiotropic regulatory gene wblA, involved in the late steps of morphogenesis. Altogether, we generated Mm overproducers with optimized growth parameters, which are useful for further genome engineering and chemoenzymatic generation of novel Mm derivatives. Analogously, such a scheme can be applied to other Streptomyces spp. MDPI 2021-12-24 /pmc/articles/PMC8778134/ /pubmed/35056478 http://dx.doi.org/10.3390/microorganisms10010030 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Makitrynskyy, Roman
Tsypik, Olga
Bechthold, Andreas
Genetic Engineering of Streptomyces ghanaensis ATCC14672 for Improved Production of Moenomycins
title Genetic Engineering of Streptomyces ghanaensis ATCC14672 for Improved Production of Moenomycins
title_full Genetic Engineering of Streptomyces ghanaensis ATCC14672 for Improved Production of Moenomycins
title_fullStr Genetic Engineering of Streptomyces ghanaensis ATCC14672 for Improved Production of Moenomycins
title_full_unstemmed Genetic Engineering of Streptomyces ghanaensis ATCC14672 for Improved Production of Moenomycins
title_short Genetic Engineering of Streptomyces ghanaensis ATCC14672 for Improved Production of Moenomycins
title_sort genetic engineering of streptomyces ghanaensis atcc14672 for improved production of moenomycins
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8778134/
https://www.ncbi.nlm.nih.gov/pubmed/35056478
http://dx.doi.org/10.3390/microorganisms10010030
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