Characterization and engineering of Streptomyces griseofuscus DSM 40191 as a potential host for heterologous expression of biosynthetic gene clusters

Abstract Streptomyces griseofuscus DSM 40191 is a fast growing Streptomyces strain that remains largely underexplored as a heterologous host. Here, we report the genome mining of S. griseofuscus, followed by the detailed exploration of its phenotype, including the production of native secondary meta...

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Autores principales:	Tetiana Gren, Christopher M. Whitford, Omkar S. Mohite, Tue S. Jørgensen, Eftychia E. Kontou, Julie B. Nielsen, Sang Yup Lee, Tilmann Weber
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/1d6bc7c1088b4caf88db11a4fa49fa8b
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spelling	oai:doaj.org-article:1d6bc7c1088b4caf88db11a4fa49fa8b2021-12-02T15:15:59ZCharacterization and engineering of Streptomyces griseofuscus DSM 40191 as a potential host for heterologous expression of biosynthetic gene clusters10.1038/s41598-021-97571-22045-2322https://doaj.org/article/1d6bc7c1088b4caf88db11a4fa49fa8b2021-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-97571-2https://doaj.org/toc/2045-2322Abstract Streptomyces griseofuscus DSM 40191 is a fast growing Streptomyces strain that remains largely underexplored as a heterologous host. Here, we report the genome mining of S. griseofuscus, followed by the detailed exploration of its phenotype, including the production of native secondary metabolites and ability to utilise carbon, nitrogen, sulphur and phosphorus sources. Furthermore, several routes for genetic engineering of S. griseofuscus were explored, including use of GusA-based vectors, CRISPR-Cas9 and CRISPR-cBEST-mediated knockouts. Two out of the three native plasmids were cured using CRISPR-Cas9 technology, leading to the generation of strain S. griseofuscus DEL1. DEL1 was further modified by the full deletion of a pentamycin BGC and an unknown NRPS BGC, leading to the generation of strain DEL2, lacking approx. 500 kbp of the genome, which corresponds to a 5.19% genome reduction. DEL2 can be characterized by faster growth and inability to produce three main native metabolites: lankacidin, lankamycin, pentamycin and their derivatives. To test the ability of DEL2 to heterologously produce secondary metabolites, the actinorhodin BGC was used. We were able to observe a formation of a blue halo, indicating a potential production of actinorhodin by both DEL2 and a wild type.Tetiana GrenChristopher M. WhitfordOmkar S. MohiteTue S. JørgensenEftychia E. KontouJulie B. NielsenSang Yup LeeTilmann WeberNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-14 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Tetiana Gren Christopher M. Whitford Omkar S. Mohite Tue S. Jørgensen Eftychia E. Kontou Julie B. Nielsen Sang Yup Lee Tilmann Weber Characterization and engineering of Streptomyces griseofuscus DSM 40191 as a potential host for heterologous expression of biosynthetic gene clusters
description	Abstract Streptomyces griseofuscus DSM 40191 is a fast growing Streptomyces strain that remains largely underexplored as a heterologous host. Here, we report the genome mining of S. griseofuscus, followed by the detailed exploration of its phenotype, including the production of native secondary metabolites and ability to utilise carbon, nitrogen, sulphur and phosphorus sources. Furthermore, several routes for genetic engineering of S. griseofuscus were explored, including use of GusA-based vectors, CRISPR-Cas9 and CRISPR-cBEST-mediated knockouts. Two out of the three native plasmids were cured using CRISPR-Cas9 technology, leading to the generation of strain S. griseofuscus DEL1. DEL1 was further modified by the full deletion of a pentamycin BGC and an unknown NRPS BGC, leading to the generation of strain DEL2, lacking approx. 500 kbp of the genome, which corresponds to a 5.19% genome reduction. DEL2 can be characterized by faster growth and inability to produce three main native metabolites: lankacidin, lankamycin, pentamycin and their derivatives. To test the ability of DEL2 to heterologously produce secondary metabolites, the actinorhodin BGC was used. We were able to observe a formation of a blue halo, indicating a potential production of actinorhodin by both DEL2 and a wild type.
format	article
author	Tetiana Gren Christopher M. Whitford Omkar S. Mohite Tue S. Jørgensen Eftychia E. Kontou Julie B. Nielsen Sang Yup Lee Tilmann Weber
author_facet	Tetiana Gren Christopher M. Whitford Omkar S. Mohite Tue S. Jørgensen Eftychia E. Kontou Julie B. Nielsen Sang Yup Lee Tilmann Weber
author_sort	Tetiana Gren
title	Characterization and engineering of Streptomyces griseofuscus DSM 40191 as a potential host for heterologous expression of biosynthetic gene clusters
title_short	Characterization and engineering of Streptomyces griseofuscus DSM 40191 as a potential host for heterologous expression of biosynthetic gene clusters
title_full	Characterization and engineering of Streptomyces griseofuscus DSM 40191 as a potential host for heterologous expression of biosynthetic gene clusters
title_fullStr	Characterization and engineering of Streptomyces griseofuscus DSM 40191 as a potential host for heterologous expression of biosynthetic gene clusters
title_full_unstemmed	Characterization and engineering of Streptomyces griseofuscus DSM 40191 as a potential host for heterologous expression of biosynthetic gene clusters
title_sort	characterization and engineering of streptomyces griseofuscus dsm 40191 as a potential host for heterologous expression of biosynthetic gene clusters
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/1d6bc7c1088b4caf88db11a4fa49fa8b
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Characterization and engineering of Streptomyces griseofuscus DSM 40191 as a potential host for heterologous expression of biosynthetic gene clusters

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