Team:UCSC/Target Organism

Yarrowia lipolytica

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  • Picking a Strain
  • General Information
    • Picking a Strain

    The yeast strain Yarrowia lipolytica was a promising species because it is classified as a “Generally Regarded as Safe” organism, found in several foods[1], used for biotechnological applications, and shown to survive on unconventional carbon sources like glycerol and oil[2]. We considered several alternative hosts but found that Y. lipolytica outperformed them all in heterologous protein expression and high transformation frequency during homologous recombination of linearized plasmids[3]. Y. lipolytica exhibits low side-product formation and high substrate specificity, which is ideal for producing a single product like progesterone[4]. The yeast also produces a fruity aroma by converting methyl ricinoleate to gamma-decalactone; we considered this to be a benefit, as our delivery method is for Y. lipolytica to be consumed orally[5].

    Saccharomyces cerevisiae is a common host organism for engineering purposes because it uses homologous recombination, a favorable DNA repair mechanism for inserting genes. Y. lipolytica has a propensity for a less favorable DNA repair mechanism, non-homologous end joining (NHEJ). To circumvent this problem, we searched for a strain that had the NHEJ mechanism, which is controlled by the Ku70 gene, knocked out. Our project also required auxotrophic markers for selective screening of our transformants, so we looked for a URA3 and LEU2 auxotroph. The paper by Verbeke et. al described the JMY2394 strain, which satisfied our parameters but was unavailable to us. We searched for a similar strain and settled on FKP393 by recommendation of Erin Bredeweg of the Pacific Northwest National Laboratory[6]. FKP393 is a NHEJ mutant derived by transformation of Po1g with the specifications matA; leu2-270; ura3; xpr2-332; axp-2; ku70::hph+[7]. This strain includes all of our specifications but lacks sufficient annotation. We were informed by Cory Schwartz of UC Riverside that this was not an issue, because the parent strain is the well-studied W29 which we can use for genome analysis. We received the FKP393 strain from the Fungal Genetics Stock Center at the Kansas State University[8].


    Yarrowia lipolytica Synthetic Pathway

    Figure 1: The pathway above shows one of the products of our strain of Yarrowia lipolytica, ergosterol, and the pathway following to produce progesterone. * Two genes, FDX1 and ADR, are also required to be inserted in the "metabolic shunt" for Y. lipolytica to produce pregnenolone.

    General Information

    Will be updated at a later date


    1. Gon calves, F. a. G., Colen, G., and Takahashi, J. A. (2014). Yarrowia lipolytica and Its Multiple Applications in the Biotechnological Industry. The Scientific World Journal
    2. Fabiszewska, A. U., Kotyrba, D., and Nowak, D. (2015). Assortment of carbon sources in medium for Yarrowia lipolytica lipase production: A statistical ap- proach. Annals of Microbiology 65, 1495–1503.
    3. Madzak, C., Gaillardin, C., and Beckerich, J.-M. (2004). Heterologous protein ex- pression and secretion in the non-conventional yeast Yarrowia lipolytica: a review. Journal of Biotechnology 109, 63–81
    4. Barth, G. (2013). Yarrowia lipolytica: Biotechnological Applications (Springer Sci- ence & Business Media). Google-Books-ID: bUS3BAAAQBAJ
    5. Aguedo, M., Gomes, N., Garcia, E. E., Wach ́e, Y., Mota, M., Teixeira, J., et al. (2005). Decalactone Production by Yarrowia lipolytica under increased O2 Trans- fer Rates. Biotechnology Letters 27, 1617–1621.
    6. Bredeweg, E. L., Pomraning, K. R., Dai, Z., Nielsen, J., Kerkhoven, E. J., and Baker, S. E. (2017). A molecular genetic toolbox for Yarrowia lipolytica. Biotechnology for Biofuels 10, 2.
    7. Madzak, C., Treton, B., and Blanchin-Roland, S. (2000). Strong Hybrid Promoters and Integrative Expression/Secretion Vectors for Quasi-Constitutive Expression of Heterologous Proteins in the Yeast Yarrowia lipolytica. Journal of Molecular Microbiology and Biotechnology 2, 207–216
    8. McClusket, K., Plamann, M., and Weist, A. (2010). The Fungal Genetics Stock Center: a repository for 50 years of fungal genetics research. Journal of Biosciences 35, 119–126.