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Updated: Jun 8, 2025

Genetic Engineering of an Unconventional Yeast for Renewable Biofuel and Biochemical Production
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Developing polycistronic expression tool in Yarrowia lipolytica.

Donghan Li1,2, Jianhui Liu1, Lingxuan Sun1

  • 1State Key Laboratory of Microbial Technology, Shandong University, Qingdao, PR China.

Synthetic and Systems Biotechnology
|November 4, 2024
PubMed
Summary

Researchers developed a new polycistronic expression tool using 2A peptides for the yeast Yarrowia lipolytica. This facilitates multi-gene expression, simplifying the construction of microbial cell factories for chemical synthesis.

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Area of Science:

  • Microbiology
  • Synthetic Biology
  • Biotechnology

Background:

  • Yarrowia lipolytica is a versatile oleaginous yeast utilized as a microbial cell factory.
  • Efficient metabolic engineering is crucial for optimizing Y. lipolytica as a cell factory.
  • Current methods for multi-gene expression in Y. lipolytica are labor-intensive.

Purpose of the Study:

  • To develop a novel polycistronic expression tool for Y. lipolytica to streamline multi-gene expression.
  • To identify efficient 2A peptides for facilitating simultaneous expression of multiple genes.
  • To demonstrate the utility of the developed tool in a relevant biosynthetic pathway.

Main Methods:

  • Comparison of various 2A peptides for cleavage efficiency in Y. lipolytica.
  • Evaluation of the impact of 2A peptides on upstream and downstream gene expression levels.
Keywords:
2A peptidesCanthaxanthinMulti-gene expression toolsYarrowia lipolytica

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  • Application of selected 2A peptides for co-expressing four genes in the canthaxanthin biosynthetic pathway.

    • Main Results:

    • Two 2A peptides, P2A and ERBV-1, were identified with high cleavage efficiency in Y. lipolytica.
    • The study determined the effects of 2A peptides on the expression of both upstream and downstream genes.
    • Successful co-expression of four genes for canthaxanthin production was achieved using a single expression cassette.

    Conclusions:

    • The developed polycistronic expression tool using 2A peptides significantly enhances multi-gene expression capabilities in Y. lipolytica.
    • This advancement simplifies and accelerates the construction of engineered Y. lipolytica cell factories.
    • The tool provides a valuable resource for metabolic engineering and the production of various compounds using Y. lipolytica.