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Redirection of lipid flux toward phospholipids in yeast increases fatty acid turnover and secretion

Bio-based production of fatty acids and fatty acid-derived products can enable sustainable substitution of petroleum-derived fuels and chemicals. However, developing new microbial cell factories for producing high levels of fatty acids requires extensive engineering of lipid metabolism, a complex an...

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Autores principales: Ferreira, Raphael, Teixeira, Paulo Gonçalves, Siewers, Verena, Nielsen, Jens
Formato: Online Artículo Texto
Lenguaje:English
Publicado: National Academy of Sciences 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5819412/
https://www.ncbi.nlm.nih.gov/pubmed/29358378
http://dx.doi.org/10.1073/pnas.1715282115
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author Ferreira, Raphael
Teixeira, Paulo Gonçalves
Siewers, Verena
Nielsen, Jens
author_facet Ferreira, Raphael
Teixeira, Paulo Gonçalves
Siewers, Verena
Nielsen, Jens
author_sort Ferreira, Raphael
collection PubMed
description Bio-based production of fatty acids and fatty acid-derived products can enable sustainable substitution of petroleum-derived fuels and chemicals. However, developing new microbial cell factories for producing high levels of fatty acids requires extensive engineering of lipid metabolism, a complex and tightly regulated metabolic network. Here we generated a Saccharomyces cerevisiae platform strain with a simplified lipid metabolism network with high-level production of free fatty acids (FFAs) due to redirected fatty acid metabolism and reduced feedback regulation. Deletion of the main fatty acid activation genes (the first step in β-oxidation), main storage lipid formation genes, and phosphatidate phosphatase genes resulted in a constrained lipid metabolic network in which fatty acid flux was directed to a large extent toward phospholipids. This resulted in simultaneous increases of phospholipids by up to 2.8-fold and of FFAs by up to 40-fold compared with wild-type levels. Further deletion of phospholipase genes PLB1 and PLB2 resulted in a 46% decrease in FFA levels and 105% increase in phospholipid levels, suggesting that phospholipid hydrolysis plays an important role in FFA production when phospholipid levels are increased. The multiple deletion mutant generated allowed for a study of fatty acid dynamics in lipid metabolism and represents a platform strain with interesting properties that provide insight into the future development of lipid-related cell factories.
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spelling pubmed-58194122018-02-21 Redirection of lipid flux toward phospholipids in yeast increases fatty acid turnover and secretion Ferreira, Raphael Teixeira, Paulo Gonçalves Siewers, Verena Nielsen, Jens Proc Natl Acad Sci U S A Biological Sciences Bio-based production of fatty acids and fatty acid-derived products can enable sustainable substitution of petroleum-derived fuels and chemicals. However, developing new microbial cell factories for producing high levels of fatty acids requires extensive engineering of lipid metabolism, a complex and tightly regulated metabolic network. Here we generated a Saccharomyces cerevisiae platform strain with a simplified lipid metabolism network with high-level production of free fatty acids (FFAs) due to redirected fatty acid metabolism and reduced feedback regulation. Deletion of the main fatty acid activation genes (the first step in β-oxidation), main storage lipid formation genes, and phosphatidate phosphatase genes resulted in a constrained lipid metabolic network in which fatty acid flux was directed to a large extent toward phospholipids. This resulted in simultaneous increases of phospholipids by up to 2.8-fold and of FFAs by up to 40-fold compared with wild-type levels. Further deletion of phospholipase genes PLB1 and PLB2 resulted in a 46% decrease in FFA levels and 105% increase in phospholipid levels, suggesting that phospholipid hydrolysis plays an important role in FFA production when phospholipid levels are increased. The multiple deletion mutant generated allowed for a study of fatty acid dynamics in lipid metabolism and represents a platform strain with interesting properties that provide insight into the future development of lipid-related cell factories. National Academy of Sciences 2018-02-06 2018-01-22 /pmc/articles/PMC5819412/ /pubmed/29358378 http://dx.doi.org/10.1073/pnas.1715282115 Text en Copyright © 2018 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by-nc-nd/4.0/ This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND) (https://creativecommons.org/licenses/by-nc-nd/4.0/) .
spellingShingle Biological Sciences
Ferreira, Raphael
Teixeira, Paulo Gonçalves
Siewers, Verena
Nielsen, Jens
Redirection of lipid flux toward phospholipids in yeast increases fatty acid turnover and secretion
title Redirection of lipid flux toward phospholipids in yeast increases fatty acid turnover and secretion
title_full Redirection of lipid flux toward phospholipids in yeast increases fatty acid turnover and secretion
title_fullStr Redirection of lipid flux toward phospholipids in yeast increases fatty acid turnover and secretion
title_full_unstemmed Redirection of lipid flux toward phospholipids in yeast increases fatty acid turnover and secretion
title_short Redirection of lipid flux toward phospholipids in yeast increases fatty acid turnover and secretion
title_sort redirection of lipid flux toward phospholipids in yeast increases fatty acid turnover and secretion
topic Biological Sciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5819412/
https://www.ncbi.nlm.nih.gov/pubmed/29358378
http://dx.doi.org/10.1073/pnas.1715282115
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