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Drop-in biofuel production using fatty acid photodecarboxylase from Chlorella variabilis in the oleaginous yeast Yarrowia lipolytica

Bruder, Stefan ; Moldenhauer, Eva Johanna ; Lemke, Robert Denis ; Ledesma-Amaro, Rodrigo ; Kabisch, Johannes (2022):
Drop-in biofuel production using fatty acid photodecarboxylase from Chlorella variabilis in the oleaginous yeast Yarrowia lipolytica. (Publisher's Version)
In: Biotechnology for Biofuels, 12, Springer Nature, ISSN 1754-6834,
DOI: 10.26083/tuprints-00013038,
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Item Type: Article
Origin: Secondary publication
Status: Publisher's Version
Title: Drop-in biofuel production using fatty acid photodecarboxylase from Chlorella variabilis in the oleaginous yeast Yarrowia lipolytica
Language: English
Abstract:

Background: Oleaginous yeasts are potent hosts for the renewable production of lipids and harbor great potential for derived products, such as biofuels. Several promising processes have been described that produce hydrocarbon drop-in biofuels based on fatty acid decarboxylation and fatty aldehyde decarbonylation. Unfortunately, besides fatty aldehyde toxicity and high reactivity, the most investigated enzyme, aldehyde-deformylating oxygenase, shows unfavorable catalytic properties which hindered high yields in previous metabolic engineering approaches.

Results: To demonstrate an alternative alkane production pathway for oleaginous yeasts, we describe the production of diesel-like, odd-chain alkanes and alkenes, by heterologously expressing a recently discovered light-driven oxidase from Chlorella variabilis (CvFAP) in Yarrowia lipolytica. Initial experiments showed that only strains engineered to have an increased pool of free fatty acids were susceptible to sufficient decarboxylation. Providing these strains with glucose and light in a synthetic medium resulted in titers of 10.9 mg/L of hydrocarbons. Using custom 3D printed labware for lighting bioreactors, and an automated pulsed glycerol fed-batch strategy, intracellular titers of 58.7 mg/L were achieved. The production of odd-numbered alkanes and alkenes with a length of 17 and 15 carbons shown in previous studies could be confirmed.

Conclusions: Oleaginous yeasts such as Yarrowia lipolytica can transform renewable resources such as glycerol into fatty acids and lipids. By heterologously expressing a fatty acid photodecarboxylase from the algae Chlorella variabilis hydrocarbons were produced in several scales from microwell plate to 400 mL bioreactors. The lighting turned out to be a crucial factor in terms of growth and hydrocarbon production, therefore, the evaluation of different conditions was an important step towards a tailor-made process. In general, the developed bioprocess shows a route to the renewable production of hydrocarbons for a variety of applications ranging from being substrates for further enzymatic or chemical modification or as a drop-in biofuel blend.

Journal or Publication Title: Biotechnology for Biofuels
Volume of the journal: 12
Publisher: Springer Nature
Collation: 13 Seiten
Classification DDC: 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie
600 Technik, Medizin, angewandte Wissenschaften > 660 Technische Chemie
Divisions: 10 Department of Biology > Computer-aided Synthetic Biology
Date Deposited: 03 Mar 2022 13:36
Last Modified: 10 Mar 2022 09:42
DOI: 10.26083/tuprints-00013038
Corresponding Links:
URN: urn:nbn:de:tuda-tuprints-130380
Additional Information:

Keywords: Drop-in biofuels, Clean fuels, Microbial biodiesel, Fatty acid photodecarboxylase, Hydrocarbons, Alkane, Oleaginous yeast, Yarrowia lipolytica

URI: https://tuprints.ulb.tu-darmstadt.de/id/eprint/13038
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