Journal article
Production of abscisic acid in the oleaginous yeast Yarrowia lipolytica
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark1
Yeast Metabolic Engineering, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark2
Department of Biotechnology and Biomedicine, Technical University of Denmark3
Section for Synthetic Biology, Department of Biotechnology and Biomedicine, Technical University of Denmark4
Yeast Biotechnology and Fermentation, Section for Synthetic Biology, Department of Biotechnology and Biomedicine, Technical University of Denmark5
DTU Fermentation Platform, Section for Synthetic Biology, Department of Biotechnology and Biomedicine, Technical University of Denmark6
Analytics, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark7
DTU Microbes Initiative, Centers, Technical University of Denmark8
Abscisic acid (ABA) is a phytohormone with applications in agriculture and human health. ABA can be produced by Botrytis cinerea, a plant pathogenic filamentous fungus. However, the cultivation process is lengthy and strain improvement by genetic engineering is difficult. Therefore, we engineered the oleaginous yeast Yarrowia lipolytica as an alternative host for ABA production.
First, we expressed five B. cinerea genes involved in ABA biosynthesis (BcABA1, BcABA2, BcABA3, BcABA4, and BcCPR1) in a Y. lipolytica chassis with optimized mevalonate flux. The strain produced 59.2 mg/L of ABA in small-scale cultivation. Next, we expressed an additional copy of each gene in the strain, but only expression of additional copy of BcABA1 gene increased the ABA titer to 168.5 mg/L.
We then integrated additional copies of mevalonate pathway and ABA-biosynthesis encoding genes, and we expressed plant ABA-transporters resulting in an improved strain producing 263.5 mg/L and 9.1 mg/g DCW ABA. Bioreactor cultivation resulted in a specific yield of 12.8 mg/g DCW ABA, however, surprisingly, the biomass level obtained in bioreactors was only 10.5 g DCW/L, with a lower ABA titer of 133.6 mg/L.
While further optimization is needed, this study confirms that Y. lipolytica as a potential alternative host for the abscisic acid production.
Language: | English |
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Publisher: | Oxford University Press |
Year: | 2022 |
ISSN: | 15671364 and 15671356 |
Types: | Journal article |
DOI: | 10.1093/femsyr/foac015 |
ORCIDs: | 0000-0003-0053-2122 , Martinez, José Luis , Rago, Daniela , Kristensen, Mette , Klitgaard, Andreas Koedfoed and Borodina, Irina |