Sterols attract increasing attention due to their important bioactivities. The oleaginous yeast Yarrowia lipolytica has large lipid droplets, which provide storage for the accumulated steroid compounds. In this study, we have successfully constructed a campesterol biosynthetic pathway by modifying the synthetic pathway of ergosterol in Y. lipolytica with different capacity of lipid synthesis. The results showed that the maximal campesterol production was produced in the engineered strain YL-D+M−E−, as the optimal lipid content. Furthermore, we found that campesterol mainly exists in the lipid droplets. The campesterol production was further accumulated through the overexpression of two copies of dhcr7. Finally, the maximal campesterol production of 837 mg/L was obtained using a 5-L bioreactor in the engineered YL-D+D+M−E−, exhibiting a 3.7-fold increase compared with the initial strain YL-D+E−. Our results demonstrate that the proper promotion of lipid content plays an important role in campesterol biosynthesis in Y. lipolytica, and what we found provides an effective strategy for the production of hydrophobic compounds.
• Campesterol was biosynthesized by deleting erg5 and introducing heterologous dhcr7.
• Campesterol production elevated via promotion of lipid content.
• Campesterol was mainly found in lipid droplets.
• Promotion of lipid content is an effective strategy to produce hydrophobic compounds.
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Data availability statement
All datasets obtained for this study are included in the manuscript/Supplementary material.
This research was supported by the National Natural Science Foundation (31972089) and National Key Research and Development Program of China (2019YFD1002402, 2019YFD100240205).
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The authors declare that they have no conflict of interest.
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Electronic supplementary material
The Supplementary material for this article can be found on the AMB website. Table S1 shows the list of primers used in this study. Table S2 shows the primers and sequences used during qPCR. Table S3 shows the nucleotide sequence of the codon-optimized dhcr7 gene. Figure S1 shows the fermentation characteristics of the initial strain YL-D+E−.
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Qian, Y.D., Tan, S.Y., Dong, G.R. et al. Increased campesterol synthesis by improving lipid content in engineered Yarrowia lipolytica. Appl Microbiol Biotechnol (2020). https://doi.org/10.1007/s00253-020-10743-4
- Lipid droplets
- Metabolic engineering
- Yarrowia lipolytica