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Optimized De Novo Eriodictyol Biosynthesis in Streptomyces albidoflavus Using an Expansion of the Golden Standard Toolkit for Its Use in Actinomycetes

Eriodictyol is a hydroxylated flavonoid displaying multiple pharmaceutical activities, such as antitumoral, antiviral or neuroprotective. However, its industrial production is limited to extraction from plants due to its inherent limitations. Here, we present the generation of a Streptomyces albidof...

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Autores principales: Magadán-Corpas, Patricia, Ye, Suhui, Pérez-Valero, Álvaro, McAlpine, Patrick L., Valdés-Chiara, Paula, Torres-Bacete, Jesús, Nogales, Juan, Villar, Claudio J., Lombó, Felipe
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10219347/
https://www.ncbi.nlm.nih.gov/pubmed/37240225
http://dx.doi.org/10.3390/ijms24108879
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author Magadán-Corpas, Patricia
Ye, Suhui
Pérez-Valero, Álvaro
McAlpine, Patrick L.
Valdés-Chiara, Paula
Torres-Bacete, Jesús
Nogales, Juan
Villar, Claudio J.
Lombó, Felipe
author_facet Magadán-Corpas, Patricia
Ye, Suhui
Pérez-Valero, Álvaro
McAlpine, Patrick L.
Valdés-Chiara, Paula
Torres-Bacete, Jesús
Nogales, Juan
Villar, Claudio J.
Lombó, Felipe
author_sort Magadán-Corpas, Patricia
collection PubMed
description Eriodictyol is a hydroxylated flavonoid displaying multiple pharmaceutical activities, such as antitumoral, antiviral or neuroprotective. However, its industrial production is limited to extraction from plants due to its inherent limitations. Here, we present the generation of a Streptomyces albidoflavus bacterial factory edited at the genome level for an optimized de novo heterologous production of eriodictyol. For this purpose, an expansion of the Golden Standard toolkit (a Type IIS assembly method based on the Standard European Vector Architecture (SEVA)) has been created, encompassing a collection of synthetic biology modular vectors (adapted for their use in actinomycetes). These vectors have been designed for the assembly of transcriptional units and gene circuits in a plug-and-play manner, as well as for genome editing using CRISPR-Cas9-mediated genetic engineering. These vectors have been used for the optimization of the eriodictyol heterologous production levels in S. albidoflavus by enhancing the flavonoid-3′-hydroxylase (F3’H) activity (by means of a chimera design) and by replacing three native biosynthetic gene clusters in the bacterial chromosome with the plant genes matBC (involved in extracellular malonate uptake and its intracellular activation into malonyl-CoA), therefore allowing more malonyl-CoA to be devoted to the heterologous production of plant flavonoids in this bacterial factory. These experiments have allowed an increase in production of 1.8 times in the edited strain (where the three native biosynthetic gene clusters have been deleted) in comparison with the wild-type strain and a 13 times increase in eriodictyol overproduction in comparison with the non-chimaera version of the F3′H enzyme.
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spelling pubmed-102193472023-05-27 Optimized De Novo Eriodictyol Biosynthesis in Streptomyces albidoflavus Using an Expansion of the Golden Standard Toolkit for Its Use in Actinomycetes Magadán-Corpas, Patricia Ye, Suhui Pérez-Valero, Álvaro McAlpine, Patrick L. Valdés-Chiara, Paula Torres-Bacete, Jesús Nogales, Juan Villar, Claudio J. Lombó, Felipe Int J Mol Sci Article Eriodictyol is a hydroxylated flavonoid displaying multiple pharmaceutical activities, such as antitumoral, antiviral or neuroprotective. However, its industrial production is limited to extraction from plants due to its inherent limitations. Here, we present the generation of a Streptomyces albidoflavus bacterial factory edited at the genome level for an optimized de novo heterologous production of eriodictyol. For this purpose, an expansion of the Golden Standard toolkit (a Type IIS assembly method based on the Standard European Vector Architecture (SEVA)) has been created, encompassing a collection of synthetic biology modular vectors (adapted for their use in actinomycetes). These vectors have been designed for the assembly of transcriptional units and gene circuits in a plug-and-play manner, as well as for genome editing using CRISPR-Cas9-mediated genetic engineering. These vectors have been used for the optimization of the eriodictyol heterologous production levels in S. albidoflavus by enhancing the flavonoid-3′-hydroxylase (F3’H) activity (by means of a chimera design) and by replacing three native biosynthetic gene clusters in the bacterial chromosome with the plant genes matBC (involved in extracellular malonate uptake and its intracellular activation into malonyl-CoA), therefore allowing more malonyl-CoA to be devoted to the heterologous production of plant flavonoids in this bacterial factory. These experiments have allowed an increase in production of 1.8 times in the edited strain (where the three native biosynthetic gene clusters have been deleted) in comparison with the wild-type strain and a 13 times increase in eriodictyol overproduction in comparison with the non-chimaera version of the F3′H enzyme. MDPI 2023-05-17 /pmc/articles/PMC10219347/ /pubmed/37240225 http://dx.doi.org/10.3390/ijms24108879 Text en © 2023 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
Magadán-Corpas, Patricia
Ye, Suhui
Pérez-Valero, Álvaro
McAlpine, Patrick L.
Valdés-Chiara, Paula
Torres-Bacete, Jesús
Nogales, Juan
Villar, Claudio J.
Lombó, Felipe
Optimized De Novo Eriodictyol Biosynthesis in Streptomyces albidoflavus Using an Expansion of the Golden Standard Toolkit for Its Use in Actinomycetes
title Optimized De Novo Eriodictyol Biosynthesis in Streptomyces albidoflavus Using an Expansion of the Golden Standard Toolkit for Its Use in Actinomycetes
title_full Optimized De Novo Eriodictyol Biosynthesis in Streptomyces albidoflavus Using an Expansion of the Golden Standard Toolkit for Its Use in Actinomycetes
title_fullStr Optimized De Novo Eriodictyol Biosynthesis in Streptomyces albidoflavus Using an Expansion of the Golden Standard Toolkit for Its Use in Actinomycetes
title_full_unstemmed Optimized De Novo Eriodictyol Biosynthesis in Streptomyces albidoflavus Using an Expansion of the Golden Standard Toolkit for Its Use in Actinomycetes
title_short Optimized De Novo Eriodictyol Biosynthesis in Streptomyces albidoflavus Using an Expansion of the Golden Standard Toolkit for Its Use in Actinomycetes
title_sort optimized de novo eriodictyol biosynthesis in streptomyces albidoflavus using an expansion of the golden standard toolkit for its use in actinomycetes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10219347/
https://www.ncbi.nlm.nih.gov/pubmed/37240225
http://dx.doi.org/10.3390/ijms24108879
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