New Promoters for Metabolic Engineering of Ashbya gossypii

Ashbya gossypii is a filamentous fungus that is currently exploited for the industrial production of riboflavin. In addition, metabolically engineered strains of A. gossypii have also been described as valuable biocatalysts for the production of different metabo...

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Autores principales:	Gloria Muñoz-Fernández, Javier-Fernando Montero-Bullón, José Luis Revuelta, Alberto Jiménez
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	<i>Ashbya gossypii</i> gene expression promoter luciferase metabolic engineering Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/03de6bf6e99b4ddbbc308be2e7dc2c17
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spelling	oai:doaj.org-article:03de6bf6e99b4ddbbc308be2e7dc2c172021-11-25T18:05:37ZNew Promoters for Metabolic Engineering of <i>Ashbya gossypii</i>10.3390/jof71109062309-608Xhttps://doaj.org/article/03de6bf6e99b4ddbbc308be2e7dc2c172021-10-01T00:00:00Zhttps://www.mdpi.com/2309-608X/7/11/906https://doaj.org/toc/2309-608X<i>Ashbya gossypii</i> is a filamentous fungus that is currently exploited for the industrial production of riboflavin. In addition, metabolically engineered strains of <i>A. gossypii</i> have also been described as valuable biocatalysts for the production of different metabolites such as folic acid, nucleosides, and biolipids. Hence, bioproduction in <i>A. gossypii</i> relies on the availability of well-performing gene expression systems both for endogenous and heterologous genes. In this regard, the identification of novel promoters, which are critical elements for gene expression, decisively helps to expand the <i>A. gossypii</i> molecular toolbox. In this work, we present an adaptation of the Dual Luciferase Reporter (DLR) Assay for promoter analysis in <i>A. gossypii</i> using integrative cassettes. We demonstrate the efficiency of the analysis through the identification of 10 new promoters with different features, including carbon source-regulatable abilities, that will highly improve the gene expression platforms used in <i>A. gossypii</i>. Three novel strong promoters (<i>P<sub>CCW12</sub></i>, <i>P<sub>SED1</sub></i>, and <i>P<sub>TSA1</sub></i>) and seven medium/weak promoters (<i>P<sub>HSP26</sub></i>, <i>P<sub>AGL366C</sub></i>, <i>P<sub>TMA10</sub></i>, <i>P<sub>CWP1</sub></i>, <i>P<sub>AFR038W</sub></i>, <i>P<sub>PFS1</sub></i>, and <i>P<sub>CDA2</sub></i>) are presented. The functionality of the promoters was further evaluated both for the overexpression and for the underexpression of the <i>A. gossypii</i> <i>MSN2</i> gene, which induced significant changes in the sporulation ability of the mutant strains.Gloria Muñoz-FernándezJavier-Fernando Montero-BullónJosé Luis RevueltaAlberto JiménezMDPI AGarticle<i>Ashbya gossypii</i>gene expressionpromoterluciferasemetabolic engineeringBiology (General)QH301-705.5ENJournal of Fungi, Vol 7, Iss 906, p 906 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	<i>Ashbya gossypii</i> gene expression promoter luciferase metabolic engineering Biology (General) QH301-705.5
spellingShingle	<i>Ashbya gossypii</i> gene expression promoter luciferase metabolic engineering Biology (General) QH301-705.5 Gloria Muñoz-Fernández Javier-Fernando Montero-Bullón José Luis Revuelta Alberto Jiménez New Promoters for Metabolic Engineering of <i>Ashbya gossypii</i>
description	<i>Ashbya gossypii</i> is a filamentous fungus that is currently exploited for the industrial production of riboflavin. In addition, metabolically engineered strains of <i>A. gossypii</i> have also been described as valuable biocatalysts for the production of different metabolites such as folic acid, nucleosides, and biolipids. Hence, bioproduction in <i>A. gossypii</i> relies on the availability of well-performing gene expression systems both for endogenous and heterologous genes. In this regard, the identification of novel promoters, which are critical elements for gene expression, decisively helps to expand the <i>A. gossypii</i> molecular toolbox. In this work, we present an adaptation of the Dual Luciferase Reporter (DLR) Assay for promoter analysis in <i>A. gossypii</i> using integrative cassettes. We demonstrate the efficiency of the analysis through the identification of 10 new promoters with different features, including carbon source-regulatable abilities, that will highly improve the gene expression platforms used in <i>A. gossypii</i>. Three novel strong promoters (<i>P<sub>CCW12</sub></i>, <i>P<sub>SED1</sub></i>, and <i>P<sub>TSA1</sub></i>) and seven medium/weak promoters (<i>P<sub>HSP26</sub></i>, <i>P<sub>AGL366C</sub></i>, <i>P<sub>TMA10</sub></i>, <i>P<sub>CWP1</sub></i>, <i>P<sub>AFR038W</sub></i>, <i>P<sub>PFS1</sub></i>, and <i>P<sub>CDA2</sub></i>) are presented. The functionality of the promoters was further evaluated both for the overexpression and for the underexpression of the <i>A. gossypii</i> <i>MSN2</i> gene, which induced significant changes in the sporulation ability of the mutant strains.
format	article
author	Gloria Muñoz-Fernández Javier-Fernando Montero-Bullón José Luis Revuelta Alberto Jiménez
author_facet	Gloria Muñoz-Fernández Javier-Fernando Montero-Bullón José Luis Revuelta Alberto Jiménez
author_sort	Gloria Muñoz-Fernández
title	New Promoters for Metabolic Engineering of <i>Ashbya gossypii</i>
title_short	New Promoters for Metabolic Engineering of <i>Ashbya gossypii</i>
title_full	New Promoters for Metabolic Engineering of <i>Ashbya gossypii</i>
title_fullStr	New Promoters for Metabolic Engineering of <i>Ashbya gossypii</i>
title_full_unstemmed	New Promoters for Metabolic Engineering of <i>Ashbya gossypii</i>
title_sort	new promoters for metabolic engineering of <i>ashbya gossypii</i>
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/03de6bf6e99b4ddbbc308be2e7dc2c17
work_keys_str_mv	AT gloriamunozfernandez newpromotersformetabolicengineeringofiashbyagossypiii AT javierfernandomonterobullon newpromotersformetabolicengineeringofiashbyagossypiii AT joseluisrevuelta newpromotersformetabolicengineeringofiashbyagossypiii AT albertojimenez newpromotersformetabolicengineeringofiashbyagossypiii
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New Promoters for Metabolic Engineering of <i>Ashbya gossypii</i>

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