Metabolic engineering of Saccharomyces cerevisiae for overproduction of triacylglycerols

Journal Article · 01 June 2018 · Metabolic Engineering Communications

DOI:https://doi.org/10.1016/j.meteno.2018.01.002· OSTI ID:1629896

Ferreira, Raphael ^[1]; Teixeira, Paulo Gonçalves ^[1]; Gossing, Michael ^[1]; David, Florian ^[1]; Siewers, Verena ^[1]; Nielsen, Jens ^[2]

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1. Chalmers University of Technology, Gothenburg, Sweden. Dept. of Biology and Biological Engineering; Chalmers University of Technology, Gothenburg, Sweden. Novo Nordisk Foundation Center for Biosustainability
2. Chalmers University of Technology, Gothenburg, Sweden. Dept. of Biology and Biological Engineering; Chalmers University of Technology, Gothenburg, Sweden. Novo Nordisk Foundation Center for Biosustainability; Technical University of Denmark, DK2800 Kgs Lyngby, Denmark. Novo Nordisk Foundation Center for Biosustainability

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, em>ACC1^S659A/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.

• Research Organization: Massachusetts Institute of Technology (MIT)
• Sponsoring Organization: USDOE
• DOE Contract Number: SC0008744
• OSTI ID: 1629896
• Journal Information: Metabolic Engineering Communications, Vol. 6, Issue C; ISSN 2214-0301
• Publisher: Elsevier
• Country of Publication: United States
• Language: English