References of "Vandermies, Marie"
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See detailInducible promoter for gene expression and synthetic biology
Nicaud, Jean-Marc; Trassaert, Marion; Thomas, Stéphane et al

Patent (in press)

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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 detailEnhancing erythritol productivity in Yarrowia lipolytica using metabolic engineering
Carly, Frédéric; Vandermies, Marie ULiege; Telek, Samuel ULiege et al

in Metabolic Engineering (2017), 42

Erythritol (1,2,3,4-butanetetrol) is a four-carbon sugar alcohol with sweetening properties that is used by the agrofood industry as a food additive. In this study, we demonstrated that metabolic ... [more ▼]

Erythritol (1,2,3,4-butanetetrol) is a four-carbon sugar alcohol with sweetening properties that is used by the agrofood industry as a food additive. In this study, we demonstrated that metabolic engineering can be used to improve the production of erythritol from glycerol in the yeast Yarrowia lipolytica. The best results were obtained using a mutant that overexpressed GUT1 and TKL1, which encode a glycerol kinase and a transketolase, respectively, and in which EYK1, which encodes erythrulose kinase, was disrupted; the latter enzyme is involved in an early step of erythritol catabolism. In this strain, erythritol productivity was 75% higher than in the wild type; furthermore, the culturing time needed to achieve maximum concentration was reduced by 40%. An additional advantage is that the strain was unable to consume the erythritol it had created, further increasing the process's efficiency. The erythritol productivity values we obtained here are among the highest reported thus far. [less ▲]

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See detailEnhancing the sweetener erythritol productivity in Yarrowia lipolytica using metabolic engineering
Carly, Frédéric; Vandermies, Marie ULiege; Nicaud, Jean-Marc et al

Conference (2017, June)

Erythritol (1,2,3,4-butanetetrol) is a four-carbon sugar alcohol used as sweeteners in food and beverage industry. It is most commonly generated from glucose via fermentation using osmophilic yeasts ... [more ▼]

Erythritol (1,2,3,4-butanetetrol) is a four-carbon sugar alcohol used as sweeteners in food and beverage industry. It is most commonly generated from glucose via fermentation using osmophilic yeasts. Yarrowia lipolytica is highly proficient at producing erythritol and can use raw glycerol instead of glucose as its main carbon source. Raw glycerol is a byproduct of the biodiesel industry, and it is a cheaper and more efficient carbon source than glucose when it comes to producing erythritol. To date, most studies seeking to improve erythritol production have used wild type strains or randomly generated mutants and have focused on optimizing the culture medium or culturing conditions. In this study, we demonstrated that metabolic engineering can be used to enhance the production of erythritol from glycerol. To this end, we constructed a set of eleven strains that overexpressed genes encoding key enzymes of either glycerol catabolism and erythritol synthesis pathway. For the best of our engineered strains, erythritol productivity in bioreactor was increased 78% relative to the wild type strain and maximum concentrations were obtained in 40% less time. [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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See detailIdentification and characterization of EYK1, a key gene for erythritol catabolism in Yarrowia lipolytica
Carly, Frédéric; Gamboa-Melendez, Heber; Vandermies, Marie ULiege et al

in Applied Microbiology and Biotechnology (2017), 101

Erythritol is a four-carbon sugar alcohol synthesized by osmophilic yeasts, such as Yarrowia lipolytica, in response to osmotic stress. This metabolite has application as food additive due to its ... [more ▼]

Erythritol is a four-carbon sugar alcohol synthesized by osmophilic yeasts, such as Yarrowia lipolytica, in response to osmotic stress. This metabolite has application as food additive due to its sweetening properties. Although Y. lipolytica can produce erythritol at a high level from glycerol, it is also able to consume it as carbon source. This ability negatively affects erythritol productivity and represents a serious drawback for the development of an efficient erythritol production process. In this study, we have isolated by insertion mutagenesis a Y. lipolytica mutant unable to grow on erythritol. Genomic characterization of the latter highlighted that the mutant phenotype is directly related to the disruption of the YALI0F01606g gene. Several experimental evidences suggested that the identified gene, renamed EYK1, encodes an erythrulose kinase. The mutant strain showed an enhanced capacity to produce erythritol as compared to the wild-type strain. Moreover, in specific experimental conditions, it is also able to convert erythritol to erythrulose, another compound of biotechnological interest. [less ▲]

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See detailDiscovering novel enzymes y functional screening of plurigenomic libraries from alga-associated Flavobacteriia and Gammaproteobacteria
Martin, Marjolaine ULiege; Vandermies, Marie ULiege; Joyeux, Coline et al

in Microbiological Research (2016), 186

Alga-associated microorganisms, in the context of their numerous interactions with the host and thecomplexity of the marine environment, are known to produce diverse hydrolytic enzymes with ... [more ▼]

Alga-associated microorganisms, in the context of their numerous interactions with the host and thecomplexity of the marine environment, are known to produce diverse hydrolytic enzymes with originalbiochemistry. We recently isolated several macroalgal-polysaccharide-degrading bacteria from the sur-face of the brown alga Ascophyllum nodosum. These active isolates belong to two classes: the Flavobacteriiaand the Gammaproteobacteria. In the present study, we constructed two “plurigenomic” (with multi-ple bacterial genomes) libraries with the 5 most interesting isolates (regarding their phylogeny andtheir enzymatic activities) of each class (Fv and Gm libraries). Both libraries were screened for diversehydrolytic activities. Five activities, out of the 48 previously identified in the natural polysaccharolyticisolates, were recovered by functional screening: a xylanase (GmXyl7), a beta-glucosidase (GmBg1), anesterase (GmEst7) and two iota-carrageenases (Fvi2.5 and Gmi1.3). We discuss here the potential role ofthe used host-cell, the average DNA insert-sizes and the used restriction enzymes on the divergent screen-ing yields obtained for both libraries and get deeper inside the “great screen anomaly”. Interestingly, thediscovered esterase probably stands for a novel family of homoserine o-acetyltransferase-like-esterases,while the two iota-carrageenases represent new members of the poorly known GH82 family (contain-ing only 19 proteins since its description in 2000). These original results demonstrate the efficiencyof our uncommon “plurigenomic” library approach and the underexplored potential of alga-associatedcultivable microbiota for the identification of novel and algal-specific enzymes. [less ▲]

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