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See detailUseful tools for genome editing in the non-conventional yeast Yarrowia lipolytica
Vandermies, Marie ULiege; Denies, Olivia ULiege; Nicaud, Jean-Marc et al

Poster (2017, September)

The non-conventional yeast Yarrowia lipolytica is increasingly used for the production of recombinant proteins or biomolecules with biotechnological or pharmaceutical applications. The development of such ... [more ▼]

The non-conventional yeast Yarrowia lipolytica is increasingly used for the production of recombinant proteins or biomolecules with biotechnological or pharmaceutical applications. The development of such a cell factory requires steps of genome editing that rely on selectable markers. The recently identified EYK1, encoding erythrulose kinase, can serve as an efficient catabolic selectable marker for genome editing in Y. lipolytica. Besides, the cloning-free strategy developed here simplifies the construction of disruption cassettes for genome editing in Y. lipolytica. [less ▲]

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See detailNew filamentous mutant of Yarrowia lipolytica and its use in biofilm bioreactors
Vandermies, Marie ULiege; Kar, Tambi ULiege; Denies, Olivia ULiege et al

Poster (2017, September)

The non-conventional yeast Yarrowia lipolytica is widely investigated for its unusual metabolic properties. Among them is the ability of Y. lipolytica to adopt an ovoid or hyphal morphology according to ... [more ▼]

The non-conventional yeast Yarrowia lipolytica is widely investigated for its unusual metabolic properties. Among them is the ability of Y. lipolytica to adopt an ovoid or hyphal morphology according to environmental conditions. The mechanism of dimorphic transition involves numerous genes, which have been poorly documented to date. Here, we report on the isolation of a filamentous mutant from an insertion mutagenesis library, the subsequent identification of the mutated gene, and the use of this filamentous mutant in biofilm bioreactors. [less ▲]

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See detailEYK1 encoding erythrulose kinase as a catabolic selectable marker for genome editing in the non-conventional yeast Yarrowia lipolytica
Vandermies, Marie ULiege; Denies, Olivia ULiege; Nicaud, Jean-Marc et al

in Journal of Microbiological Methods (2017), 139

We report here on EYK1, encoding erythrulose kinase, as an efficient catabolic selectable marker for genome editing in Y. lipolytica. Compared to auxotrophic markers, EYK1 increases the growth rate of ... [more ▼]

We report here on EYK1, encoding erythrulose kinase, as an efficient catabolic selectable marker for genome editing in Y. lipolytica. Compared to auxotrophic markers, EYK1 increases the growth rate of transformants and allows improved efficiency of transformation. The utility of the marker EYK1 in a replicative vector was also demonstrated. [less ▲]

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See detailNew inducible promoter for gene expression and synthetic biology in Yarrowia lipolytica
Trassaert, Marion; Vandermies, Marie ULiege; Carly, Frédéric et al

in Microbial Cell Factories (2017), 16(141),

Background The oleaginous yeast Yarrowia lipolytica is increasingly used as alternative cell factory for the production of recombinant proteins. At present, several promoters with different strengths have ... [more ▼]

Background The oleaginous yeast Yarrowia lipolytica is increasingly used as alternative cell factory for the production of recombinant proteins. At present, several promoters with different strengths have been developed based either on the constitutive pTEF promoter or on oleic acid inducible promoters such as pPOX2 and pLIP2. Although these promoters are highly efficient, there is still a lack of versatile inducible promoters for gene expression in Y. lipolytica. Results We have isolated and characterized the promoter of the EYK1 gene coding for an erythrulose kinase. pEYK1 induction was found to be impaired in media supplemented with glucose and glycerol, while the presence of erythritol and erythrulose strongly increased the promoter induction level. Promoter characterization and mutagenesis allowed the identification of the upstream activating sequence UAS1EYK1. New hybrid promoters containing tandem repeats of either UAS1XPR2 or UAS1EYK1 were developed showing higher expression levels than the native pEYK1 promoter. Furthermore, promoter strength was improved in a strain carrying a deletion in the EYK1 gene, allowing thus the utilization of erythritol and erythrulose as free inducer. Conclusions Novel tunable and regulated promoters with applications in the field of heterologous protein production, metabolic engineering, and synthetic biology have been developed, thus filling the gap of the absence of versatile inducible promoter in the yeast Y. lipolytica. [less ▲]

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