Increasing carotenoid production in Xanthophyllomyces dendrorhous/Phaffia rhodozyma: SREBP pathway activation and promoter engineering
Abstract The yeast Xanthophyllomyces dendrorhous synthesizes astaxanthin, a high-value carotenoid with biotechnological relevance in the nutraceutical and aquaculture industries. However, enhancing carotenoid production through strain engineering remains an ongoing challenge. Recent studies have dem...
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2024-11-01
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| Series: | Biological Research |
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| Online Access: | https://doi.org/10.1186/s40659-024-00559-1 |
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| author | Alejandro Durán Maximiliano Venegas Salvador Barahona Dionisia Sepúlveda Marcelo Baeza Víctor Cifuentes Jennifer Alcaíno |
| author_facet | Alejandro Durán Maximiliano Venegas Salvador Barahona Dionisia Sepúlveda Marcelo Baeza Víctor Cifuentes Jennifer Alcaíno |
| author_sort | Alejandro Durán |
| collection | DOAJ |
| description | Abstract The yeast Xanthophyllomyces dendrorhous synthesizes astaxanthin, a high-value carotenoid with biotechnological relevance in the nutraceutical and aquaculture industries. However, enhancing carotenoid production through strain engineering remains an ongoing challenge. Recent studies have demonstrated that carotenogenesis in X. dendrorhous is regulated by the SREBP pathway, which includes the transcription factor Sre1, particularly in the mevalonate pathway that also produces precursors used for ergosterol synthesis. In this study, we explored a novel approach to enhance carotenoid synthesis by replacing the native crtE promoter, which drives geranylgeranyl pyrophosphate synthesis (the step where carotenogenesis diverges from ergosterol biosynthesis), with the promoter of the HMGS gene, which encodes 3-hydroxy-3-methylglutaryl-CoA synthase from the mevalonate pathway. The impact of this substitution was evaluated in two mutant strains that already overproduce carotenoids due to the presence of an active Sre1 transcription factor: CBS.cyp61-, which does not produce ergosterol and strain CBS.SRE1N.FLAG, which constitutively expresses the active form of Sre1. Wild-type strain CBS6938 was used as a control. Our results showed that this modification increased the crtE transcript levels more than threefold and fourfold in CBS.cyp61 − .pHMGS/crtE and CBS.SRE1N.FLAG.pHMGS/crtE, respectively, resulting in 1.43-fold and 1.22-fold increases in carotenoid production. In contrast, this modification did not produce significant changes in the wild-type strain, which lacks the active Sre1 transcription factor under the same culture conditions. This study highlights the potential of promoter substitution strategies involving genes regulated by Sre1 to enhance carotenoid production, specifically in strains where the SREBP pathway is activated, offering a promising avenue for strain improvement in industrial applications. |
| format | Article |
| id | doaj-art-aa3455e8fc3b455289b1a5e2dc1c7d88 |
| institution | Kabale University |
| issn | 0717-6287 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Biological Research |
| spelling | doaj-art-aa3455e8fc3b455289b1a5e2dc1c7d882024-11-10T12:08:47ZengBMCBiological Research0717-62872024-11-0157111310.1186/s40659-024-00559-1Increasing carotenoid production in Xanthophyllomyces dendrorhous/Phaffia rhodozyma: SREBP pathway activation and promoter engineeringAlejandro Durán0Maximiliano Venegas1Salvador Barahona2Dionisia Sepúlveda3Marcelo Baeza4Víctor Cifuentes5Jennifer Alcaíno6Departamento de Ciencias Ecológicas, Facultad de Ciencias, Universidad de ChileDepartamento de Ciencias Ecológicas, Facultad de Ciencias, Universidad de Chile Facultad de Ciencias, Universidad de Chile Facultad de Ciencias, Universidad de ChileDepartamento de Ciencias Ecológicas, Facultad de Ciencias, Universidad de ChileDepartamento de Ciencias Ecológicas, Facultad de Ciencias, Universidad de ChileDepartamento de Ciencias Ecológicas, Facultad de Ciencias, Universidad de ChileAbstract The yeast Xanthophyllomyces dendrorhous synthesizes astaxanthin, a high-value carotenoid with biotechnological relevance in the nutraceutical and aquaculture industries. However, enhancing carotenoid production through strain engineering remains an ongoing challenge. Recent studies have demonstrated that carotenogenesis in X. dendrorhous is regulated by the SREBP pathway, which includes the transcription factor Sre1, particularly in the mevalonate pathway that also produces precursors used for ergosterol synthesis. In this study, we explored a novel approach to enhance carotenoid synthesis by replacing the native crtE promoter, which drives geranylgeranyl pyrophosphate synthesis (the step where carotenogenesis diverges from ergosterol biosynthesis), with the promoter of the HMGS gene, which encodes 3-hydroxy-3-methylglutaryl-CoA synthase from the mevalonate pathway. The impact of this substitution was evaluated in two mutant strains that already overproduce carotenoids due to the presence of an active Sre1 transcription factor: CBS.cyp61-, which does not produce ergosterol and strain CBS.SRE1N.FLAG, which constitutively expresses the active form of Sre1. Wild-type strain CBS6938 was used as a control. Our results showed that this modification increased the crtE transcript levels more than threefold and fourfold in CBS.cyp61 − .pHMGS/crtE and CBS.SRE1N.FLAG.pHMGS/crtE, respectively, resulting in 1.43-fold and 1.22-fold increases in carotenoid production. In contrast, this modification did not produce significant changes in the wild-type strain, which lacks the active Sre1 transcription factor under the same culture conditions. This study highlights the potential of promoter substitution strategies involving genes regulated by Sre1 to enhance carotenoid production, specifically in strains where the SREBP pathway is activated, offering a promising avenue for strain improvement in industrial applications.https://doi.org/10.1186/s40659-024-00559-1AstaxanthinGene expression regulationMetabolic engineeringMevalonate pathwaySREBP/Sre1 transcription factorPromoter replacement |
| spellingShingle | Alejandro Durán Maximiliano Venegas Salvador Barahona Dionisia Sepúlveda Marcelo Baeza Víctor Cifuentes Jennifer Alcaíno Increasing carotenoid production in Xanthophyllomyces dendrorhous/Phaffia rhodozyma: SREBP pathway activation and promoter engineering Biological Research Astaxanthin Gene expression regulation Metabolic engineering Mevalonate pathway SREBP/Sre1 transcription factor Promoter replacement |
| title | Increasing carotenoid production in Xanthophyllomyces dendrorhous/Phaffia rhodozyma: SREBP pathway activation and promoter engineering |
| title_full | Increasing carotenoid production in Xanthophyllomyces dendrorhous/Phaffia rhodozyma: SREBP pathway activation and promoter engineering |
| title_fullStr | Increasing carotenoid production in Xanthophyllomyces dendrorhous/Phaffia rhodozyma: SREBP pathway activation and promoter engineering |
| title_full_unstemmed | Increasing carotenoid production in Xanthophyllomyces dendrorhous/Phaffia rhodozyma: SREBP pathway activation and promoter engineering |
| title_short | Increasing carotenoid production in Xanthophyllomyces dendrorhous/Phaffia rhodozyma: SREBP pathway activation and promoter engineering |
| title_sort | increasing carotenoid production in xanthophyllomyces dendrorhous phaffia rhodozyma srebp pathway activation and promoter engineering |
| topic | Astaxanthin Gene expression regulation Metabolic engineering Mevalonate pathway SREBP/Sre1 transcription factor Promoter replacement |
| url | https://doi.org/10.1186/s40659-024-00559-1 |
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