Please use this identifier to cite or link to this item: doi:10.22028/D291-41076
Title: Multi-omics view of recombinant Yarrowia lipolytica: Enhanced ketogenic amino acid catabolism increases polyketide-synthase-driven docosahexaenoic production to high selectivity at the gram scale
Author(s): Jovanovic Gasovic, Sofija
Dietrich, Demian
Gläser, Lars
Cao, Peng
Kohlstedt, Michael
Wittmann, Christoph
Language: English
Title: Metabolic Engineering
Volume: 80
Pages: 45-65
Publisher/Platform: Elsevier
Year of Publication: 2023
Free key words: PUFA
Omega-3 fatty acid
Docosahexaenoic acid
DHA
Yarrowia lipolytica
Fatty acid
CoA thioester
Acetyl-CoA
Malonyl-CoA
PKS synthase
Glycerol
Ketogenic amino acid
Transcriptome
Metabolome
Fluxome
13C
Systems biology
Multi-omics
Polyketide
Lipid
DDC notations: 500 Science
Publikation type: Journal Article
Abstract: DHA is a marine PUFA of commercial value, given its multiple health benefits. The worldwide emerging shortage in DHA supply has increased interest in microbial cell factories that can provide the compound de novo. In this regard, the present work aimed to improve DHA production in the oleaginous yeast strain Y. lipolytica Af4, which synthetized the PUFA via a heterologous myxobacterial polyketide synthase (PKS)-like gene cluster. As starting point, we used transcriptomics, metabolomics, and 13C-based metabolic pathway profiling to study the cellular dynamics of Y. lipolytica Af4. The shift from the growth to the stationary DHA-production phase was associated with fundamental changes in carbon core metabolism, including a strong upregulation of the PUFA gene cluster, as well as an increase in citrate and fatty acid degradation. At the same time, the intracellular levels of the two DHA precursors acetyl-CoA and malonyl-CoA dropped by up to 98% into the picomolar range. Interestingly, the degradation pathways for the ketogenic amino acids L-lysine, L-leucine, and L-isoleucine were transcriptionally activated, presumably to provide extra acetyl-CoA. Supplementation with small amounts of these amino acids at the beginning of the DHA production phase beneficially increased the intracellular CoA-ester pools and boosted the DHA titer by almost 40%. Isotopic 13C-tracer studies revealed that the supplements were efficiently directed toward intracellular CoA-esters and DHA. Hereby, L-lysine was found to be most efficient, as it enabled long-term activation, due to storage within the vacuole and continuous breakdown. The novel strategy enabled DHA production in Y. lipolytica at the gram scale for the first time. DHA was produced at a high selectivity (27% of total fatty acids) and free of the structurally similar PUFA DPA, which facilitates purification for high-value medical applications that require API-grade DHA. The assembled multi-omics picture of the central metabolism of Y. lipolytica provides valuable insights into this important yeast. Beyond our work, the enhanced catabolism of ketogenic amino acids seems promising for the overproduction of other compounds in Y. lipolytica, whose synthesis is limited by the availability of CoA ester precursors.
DOI of the first publication: 10.1016/j.ymben.2023.09.003
URL of the first publication: https://doi.org/10.1016/j.ymben.2023.09.003
Link to this record: urn:nbn:de:bsz:291--ds-410763
hdl:20.500.11880/36863
http://dx.doi.org/10.22028/D291-41076
ISSN: 1096-7176
Date of registration: 14-Nov-2023
Description of the related object: Supplementary data
Related object: https://ars.els-cdn.com/content/image/1-s2.0-S1096717623001234-mmc1.pdf
Faculty: NT - Naturwissenschaftlich- Technische Fakultät
Department: NT - Biowissenschaften
Professorship: NT - Prof. Dr. Christoph Wittmann
Collections:SciDok - Der Wissenschaftsserver der Universität des Saarlandes

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