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Promoters for lipogenesis-specific downregulation in Yarrowia lipolytica.

ABSTRACT: Yarrowia lipolytica is a non-conventional yeast with potential applications in the biofuel and biochemical industries. It is an oleaginous yeast that accumulates lipids when it encounters nutrient limitation in the presence of excess carbon. Its molecular toolbox includes promoters for robust constitutive expression, regulated expression through the addition of media components and inducible expression during lipid accumulation. To date, no promoters have been identified that lead to downregulation at the transition from growth to lipid accumulation. We identified four native Y. lipolytica promoters that downregulate the expression of genes at this natural transition. Using the fatty acid desaturase genes FAD2 and OLE1 as reporter genes for these promoters, we correlated repression of desaturase transcript levels with a reduction of desaturated fatty acids at the transition to lipid accumulation. These promoters can restrict to the growth phase an essential or favorable activity that is undesirable during lipid accumulation under traditional fermentation conditions without media additions. This expression pattern results in lipogenesis phase-specific changes that could be useful in applications relating to optimizing lipid yield and composition.

SUBMITTER: Kamineni A

PROVIDER: S-EPMC7335367 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BackgroundProtein secretion is a universal cellular process involving vesicles which bud and fuse between organelles to bring proteins to their final destination. Vesicle budding is mediated by protein coats; vesicle targeting and fusion depend on Rab GTPase, tethering factors and SNARE complexes. The Génolevures II sequencing project made available entire genome sequences of four hemiascomycetous yeasts, Yarrowia lipolytica, Debaryomyces hansenii, Kluyveromyces lactis and Candida glabrata. Y. lipolytica is a dimorphic yeast and has good capacities to secrete proteins. The translocation of nascent protein through the endoplasmic reticulum membrane was well studied in Y. lipolytica and is largely co-translational as in the mammalian protein secretion pathway.ResultsWe identified S. cerevisiae proteins involved in vesicular secretion and these protein sequences were used for the BLAST searches against Génolevures protein database (Y. lipolytica, C. glabrata, K. lactis and D. hansenii). These proteins are well conserved between these yeasts and Saccharomyces cerevisiae. We note several specificities of Y. lipolytica which may be related to its good protein secretion capacities and to its dimorphic aspect. An expansion of the Y. lipolytica Rab protein family was observed with autoBLAST and the Rab2- and Rab4-related members were identified with BLAST against NCBI protein database. An expansion of this family is also found in filamentous fungi and may reflect the greater complexity of the Y. lipolytica secretion pathway. The Rab4p-related protein may play a role in membrane recycling as rab4 deleted strain shows a modification of colony morphology, dimorphic transition and permeability. Similarly, we find three copies of the gene (SSO) encoding the plasma membrane SNARE protein. Quantification of the percentages of proteins with the greatest homology between S. cerevisiae, Y. lipolytica and animal homologues involved in vesicular transport shows that 40% of Y. lipolytica proteins are closer to animal ones, whereas they are only 13% in the case of S. cerevisiae.ConclusionThese results provide further support for the idea, previously noted about the endoplasmic reticulum translocation pathway, that Y. lipolytica is more representative of vesicular secretion of animals and other fungi than is S. cerevisiae.

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Promoters for lipogenesis-specific downregulation in Yarrowia lipolytica.

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