New Promoters for Metabolic Engineering of <i>Ashbya gossypii</i>
<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 metabo...
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| Autores principales: | , , , |
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| Formato: | article |
| Lenguaje: | EN |
| Publicado: |
MDPI AG
2021
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| Materias: | |
| Acceso en línea: | https://doaj.org/article/03de6bf6e99b4ddbbc308be2e7dc2c17 |
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| Sumario: | <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. |
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