Skip to main content

Advertisement

SpringerLink
Log in

New pAraDH2 Promoter for Metabolic Engineering of the Yarrowia lipolytica Yeast

  • PRODUCERS, BIOLOGY, SELECTION, AND GENE ENGINEERING
  • Published:
Applied Biochemistry and Microbiology Aims and scope Submit manuscript

Abstract

The ability to utilize a number of polyhydric alcohols as a sole carbon source has been studied in the Yarrowia lipolytica yeast. The efficiency of the promoter of the Y. lipolytica native AraDH2 gene encoding the enzyme D-mannitol/D-arabitol dehydrogenase was assessed during yeast growth on a minimal medium with different carbon sources. For this purpose, the promoter region of the AraDH2 gene was transcriptionally fused with the green fluorescent protein hrGFP gene, and the construct was used to transform Y. lipolytica. A β-carotene producing strain of Y. lipolytica was created using the described promoter; the strain carried the Mucor circinelloides CarRP and CarB genes encoding the bifunctional enzyme phytoene synthase/lycopene β-cyclase (CarRP) and phytoene dehydrogenase (CarB) as well as the GGPPSs7 gene Synechococcus sp. geranylgeranyl pyrophosphate synthase. The nucleotide sequence encoding the fused CarRP and GGPPSs7 under the regulation of the pAraDH2 promoter and the CarB gene under the control of the pTEF promoter were introduced into the yeast genome. As a result, a transformant was obtained capable of producing 66.3, 121.2, and 148.9 mg/L of β-carotene after 5 days of cultivation in test tubes on media containing glycerol, sucrose, and glucose, respectively. The obtained results testify to the high potential of the pAraDH2 promoter in the field of genetic engineering.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1.
Fig. 2.
Fig. 3.
Fig. 4.

REFERENCES

  1. Abdel-Mawgoud, A.M., Markham, K.A., Palmer, C.M., Liu, N., Stephanopoulos, G., and Alper, H.S., Metabolic engineering in the host Yarrowia lipolytica, Metab. Eng., 2018, vol. 50, pp. 192–208. https://doi.org/10.1016/j.ymben.2018.07.016

    Article  CAS  PubMed  Google Scholar 

  2. Miller, K.K. and Alper, H.S., Yarrowia lipolytica: more than an oleaginous workhorse, Appl. Microbiol. Biotechnol., 2019, vol. 103, pp. 9251– 9262. https://doi.org/10.1007/s00253-019-10200-x

    Article  CAS  PubMed  Google Scholar 

  3. Barth, G. and Gaillardin, C., Physiology and genetics of the dimorphic fungus Yarrowia lipolytica, FEMS Microbiol. Rev., 1997, vol. 19, no. 4, pp. 219–237. https://doi.org/10.1111/j.1574-6976.1997.tb00299.x

    Article  CAS  PubMed  Google Scholar 

  4. Ledesma-Amaro, R. and Nicaud, J.M., Metabolic engineering for expanding the substrate range of Yarrowia lipolytica, Trends Biotechnol., 2016, vol. 34, no. 10, pp. 798–809. https://doi.org/10.1016/j.tibtech.2016.04.010

    Article  CAS  PubMed  Google Scholar 

  5. Larroude, M., Rossignol, T., Nicaud, J.M., and Ledesma-Amaro, R., Synthetic biology tools for engineering Yarrowia lipolytica, Biotechnol. Adv., 2018, vol. 36, no. 8, pp. 2150–2164. https://doi.org/10.1016/j.biotechadv.2018.10.004

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Poorinmohammad, N. and Kerkhoven, E.J., Systems-level approaches for understanding and engineering of the oleaginous cell factory Yarrowia lipolytica, Biotechnol. Bioeng., 2021, vol. 118, no. 10, pp. 3640–3654. https://doi.org/10.1002/bit.27859

    Article  CAS  PubMed  Google Scholar 

  7. Zhang, Y., Nielsen, J., and Liu, Z., Engineering yeast metabolism for production of terpenoids for use as perfume ingredients, pharmaceuticals and biofuels, FEMS Yeast Res., 2017, vol. 17, no. 8. https://doi.org/10.1093/femsyr/fox080

  8. Gao, S., Tong, Y., Zhu, L., Ge, M., Zhang, Y., Chen, D., Jiang, Y., and Yang, Sh., Iterative integration of multiple-copy pathway genes in Yarrowia lipolytica for heterologous beta-carotene production, Metab. Eng., 2017, vol. 41, pp. 192–201. https://doi.org/10.1016/j.ymben.2017.04.004

    Article  CAS  PubMed  Google Scholar 

  9. Larroude, M., Celinska, E., Back, A., Thomas, S., Nicaud, J.-M., and Ledesma-Amaro, R., A synthetic biology approach to transform Yarrowia lipolytica into a competitive biotechnological producer of β-carotene, Biotechnol. Bioeng., 2018, no. 2, pp. 464–472. https://doi.org/10.1002/bit.26473

  10. Wang, N., Chi, P., Zou, Y., Xu, Y., Xu, S., Bilal, M., Fickers, P., and Cheng, H., Metabolic engineering of Yarrowia lipolytica for thermoresistance and enhanced erythritol productivity, Biotechnol. Biofuels, 2020, vol. 13, p. 176. https://doi.org/10.1186/s13068-020-01815-8

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Sambrook, J., Maniatis, T., and Fritsch, E., Molecular Cloning: Laboratory Mannual, New York: Cold Spring Harbor Laboratory Press, 1989, 2nd ed.

    Google Scholar 

  12. Yuzbashev, T., Yuzbasheva, E., Melkina, O., Patel, D., Bubnov, D., Dietz, H., and Ledesma-Amaro, R., A DNA assembly toolkit to unlock the CRISPR/Cas9 potential for metabolic engineering, 2022. www.researchsquare.com/article/rs-1570357/v1.

  13. Blazeck, J., Liu, L., Redden, H., and Alper, H., Tuning gene expression in Yarrowia lipolytica by a hybrid promoter approach, Appl. Environ. Microbiol., 2011, vol. 77, no. 22, pp. 7905–7914. https://doi.org/10.1128/AEM.05763-11

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Sabra, W., Bommareddy, R.R., Maheshwari, G., Papanikolaou, S., and Zeng, A.-P., Substrates and oxygen dependent citric acid production by Yarrowia lipolytica: insights through transcriptome and fluxome analyses, Microb. Cell. Fact., 2017, vol. 16, no. 1, p. 78. https://doi.org/10.1186/s12934-017-0690-0

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Taratynova, M.O., Kosikhina, Y.M., Vinogradova, E.B., Dementev, D.A., Korobov, V.S., Zolottsev, V.A., Yuzbashev, T.V., Yuzbasheva, E.Y., and Sineoky, S.P., Accumulation of neutral lipids and β-carotene by the Yarrowia lipolytica yeast on a medium with sucrose as a carbon source, Biotechnology, 2021, vol. 37, pp. 29–41. https://doi.org/10.21519/0234-2758-2021-37-3-29-41

    Article  Google Scholar 

  16. Mohandesi, N., Siadat, S.O., Haghbeen, K., and Hesampour, A., Cloning and expression of Saccharomyces cerevisiae SUC2 gene in yeast platform and characterization of recombinant enzyme biochemical properties, 3 Biotech, 2016, vol. 6, no. 2, p. 129. https://doi.org/10.1007/s13205-016-0441-7

Download references

Funding

This work was supported by State Assignment of National Research Center Kurchatov Institute.

Author information

Authors and Affiliations

  1. Kurchatov Institute National Research Center, 123182, Moscow, Russia

    M. O. Taratynova, Iu. M. Fediayeva, D. A. Dementiev, O. E. Melkina & S. P. Sineoky

  2. Plant Sciences and the Bioeconomy, AL5 2JQ, Harpenden, Rothamsted Research, West Common, United Kingdom

    T. V. Yuzbashev

  3. BioMediCan Inc., 94538, Fremont, CA, United States

    E. Y. Yuzbasheva

  4. BioKai Inc., 94538, Fremont, CA, United States

    E. Y. Yuzbasheva

Authors
  1. M. O. Taratynova
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Iu. M. Fediayeva
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. D. A. Dementiev
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. O. E. Melkina
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. T. V. Yuzbashev
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. S. P. Sineoky
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. E. Y. Yuzbasheva
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to M. O. Taratynova.

Ethics declarations

ETHICS APPROVAL AND CONSENT TO PARTICIPATE

This work does not contain any studies involving human and animal subjects.

CONFLICT OF INTEREST

The authors of this work declare that they have no conflicts of interest.

Additional information

Publisher’s Note.

Pleiades Publishing remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Abbreviations: (in alphabetical order) AraDH2—D-mannitol/D-arabitol dehydrogenase; CarB—phytoene dehydrogenase; CarRP—bifunctional phytoene synthase/lycopene β-cyclase; GGPPs7—geranylgeranyl pyrophosphate synthase; hrGFP—green fluorescent protein; ScSUC2—invertase.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Taratynova, M.O., Fediayeva, I.M., Dementiev, D.A. et al. New pAraDH2 Promoter for Metabolic Engineering of the Yarrowia lipolytica Yeast. Appl Biochem Microbiol 59, 1157–1167 (2023). https://doi.org/10.1134/S0003683823090120

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S0003683823090120

Keywords:

Search

Navigation