Metabolic engineering of Saccharomyces cerevisiae for overproduction of triacylglycerols
Triacylglycerols (TAGs) are valuable versatile compounds that can be used as metabolites for nutrition and health, as well as feedstocks for biofuel production. Although Saccharomyces cerevisiae is the favored microbial cell factory for industrial production of biochemicals, it does not produce larg...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2018-06-01
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| Series: | Metabolic Engineering Communications |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S221403011730024X |
| _version_ | 1818081972988149760 |
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| author | Raphael Ferreira Paulo Gonçalves Teixeira Michael Gossing Florian David Verena Siewers Jens Nielsen |
| author_facet | Raphael Ferreira Paulo Gonçalves Teixeira Michael Gossing Florian David Verena Siewers Jens Nielsen |
| author_sort | Raphael Ferreira |
| collection | DOAJ |
| description | Triacylglycerols (TAGs) are valuable versatile compounds that can be used as metabolites for nutrition and health, as well as feedstocks for biofuel production. Although Saccharomyces cerevisiae is the favored microbial cell factory for industrial production of biochemicals, it does not produce large amounts of lipids and TAGs comprise only ~1% of its cell dry weight. Here, we engineered S. cerevisiae to reorient its metabolism for overproduction of TAGs, by regulating lipid droplet associated-proteins involved in TAG synthesis and hydrolysis. We implemented a push-and-pull strategy by overexpressing genes encoding a deregulated acetyl-CoA carboxylase, ACC1S659A/S1157A (ACC1**), as well as the last two steps of TAG formation: phosphatidic phosphatase (PAH1) and diacylglycerol acyltransferase (DGA1), ultimately leading to 129 mg∙gCDW−1 of TAGs. Disruption of TAG lipase genes TGL3, TGL4, TGL5 and sterol acyltransferase gene ARE1 increased the TAG content to 218 mg∙gCDW−1. Further disruption of the beta-oxidation by deletion of POX1, as well as glycerol-3-phosphate utilization through deletion of GUT2, did not affect TAGs levels. Finally, disruption of the peroxisomal fatty acyl-CoA transporter PXA1 led to accumulation of 254 mg∙gCDW−1. The TAG levels achieved here are the highest titer reported in S. cerevisiae, reaching 27.4% of the maximum theoretical yield in minimal medium with 2% glucose. This work shows the potential of using an industrially established and robust yeast species for high level lipid production. |
| first_indexed | 2024-12-10T19:14:42Z |
| format | Article |
| id | doaj.art-c5a79a1d9c6842708848dbc949e60b35 |
| institution | Directory Open Access Journal |
| issn | 2214-0301 |
| language | English |
| last_indexed | 2024-12-10T19:14:42Z |
| publishDate | 2018-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Metabolic Engineering Communications |
| spelling | doaj.art-c5a79a1d9c6842708848dbc949e60b352022-12-22T01:36:37ZengElsevierMetabolic Engineering Communications2214-03012018-06-0162227Metabolic engineering of Saccharomyces cerevisiae for overproduction of triacylglycerolsRaphael Ferreira0Paulo Gonçalves Teixeira1Michael Gossing2Florian David3Verena Siewers4Jens Nielsen5Department of Biology and Biological Engineering, Chalmers University of Technology, SE412 96 Gothenburg, Sweden; Novo Nordisk Foundation Center for Biosustainability, Chalmers University of Technology, SE412 96 Gothenburg, SwedenDepartment of Biology and Biological Engineering, Chalmers University of Technology, SE412 96 Gothenburg, Sweden; Novo Nordisk Foundation Center for Biosustainability, Chalmers University of Technology, SE412 96 Gothenburg, SwedenDepartment of Biology and Biological Engineering, Chalmers University of Technology, SE412 96 Gothenburg, Sweden; Novo Nordisk Foundation Center for Biosustainability, Chalmers University of Technology, SE412 96 Gothenburg, SwedenDepartment of Biology and Biological Engineering, Chalmers University of Technology, SE412 96 Gothenburg, Sweden; Novo Nordisk Foundation Center for Biosustainability, Chalmers University of Technology, SE412 96 Gothenburg, SwedenDepartment of Biology and Biological Engineering, Chalmers University of Technology, SE412 96 Gothenburg, Sweden; Novo Nordisk Foundation Center for Biosustainability, Chalmers University of Technology, SE412 96 Gothenburg, SwedenDepartment of Biology and Biological Engineering, Chalmers University of Technology, SE412 96 Gothenburg, Sweden; Novo Nordisk Foundation Center for Biosustainability, Chalmers University of Technology, SE412 96 Gothenburg, Sweden; Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, DK2800 Kgs Lyngby, Denmark; Corresponding author at: Department of Biology and Biological Engineering, Chalmers University of Technology, SE412 96 Gothenburg, Sweden.Triacylglycerols (TAGs) are valuable versatile compounds that can be used as metabolites for nutrition and health, as well as feedstocks for biofuel production. Although Saccharomyces cerevisiae is the favored microbial cell factory for industrial production of biochemicals, it does not produce large amounts of lipids and TAGs comprise only ~1% of its cell dry weight. Here, we engineered S. cerevisiae to reorient its metabolism for overproduction of TAGs, by regulating lipid droplet associated-proteins involved in TAG synthesis and hydrolysis. We implemented a push-and-pull strategy by overexpressing genes encoding a deregulated acetyl-CoA carboxylase, ACC1S659A/S1157A (ACC1**), as well as the last two steps of TAG formation: phosphatidic phosphatase (PAH1) and diacylglycerol acyltransferase (DGA1), ultimately leading to 129 mg∙gCDW−1 of TAGs. Disruption of TAG lipase genes TGL3, TGL4, TGL5 and sterol acyltransferase gene ARE1 increased the TAG content to 218 mg∙gCDW−1. Further disruption of the beta-oxidation by deletion of POX1, as well as glycerol-3-phosphate utilization through deletion of GUT2, did not affect TAGs levels. Finally, disruption of the peroxisomal fatty acyl-CoA transporter PXA1 led to accumulation of 254 mg∙gCDW−1. The TAG levels achieved here are the highest titer reported in S. cerevisiae, reaching 27.4% of the maximum theoretical yield in minimal medium with 2% glucose. This work shows the potential of using an industrially established and robust yeast species for high level lipid production.http://www.sciencedirect.com/science/article/pii/S221403011730024X |
| spellingShingle | Raphael Ferreira Paulo Gonçalves Teixeira Michael Gossing Florian David Verena Siewers Jens Nielsen Metabolic engineering of Saccharomyces cerevisiae for overproduction of triacylglycerols Metabolic Engineering Communications |
| title | Metabolic engineering of Saccharomyces cerevisiae for overproduction of triacylglycerols |
| title_full | Metabolic engineering of Saccharomyces cerevisiae for overproduction of triacylglycerols |
| title_fullStr | Metabolic engineering of Saccharomyces cerevisiae for overproduction of triacylglycerols |
| title_full_unstemmed | Metabolic engineering of Saccharomyces cerevisiae for overproduction of triacylglycerols |
| title_short | Metabolic engineering of Saccharomyces cerevisiae for overproduction of triacylglycerols |
| title_sort | metabolic engineering of saccharomyces cerevisiae for overproduction of triacylglycerols |
| url | http://www.sciencedirect.com/science/article/pii/S221403011730024X |
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