References of "Denies, Olivia"
     in
Bookmark and Share    
See detailBy-products show potential in glutathione production by engineered non-conventional yeast Yarrowia lipolytica
Do Thi Hoang Diem, ULiege; Steels, Sébastien ULiege; Denies, Olivia ULiege et al

Poster (2019, May 08)

Strong constitutive promoter pTEF were used to overexpress genes encoding the two enzymes (L-cysteine gamma-ligase and glutathione synthase) which catalyze the synthesis of GSH in Yarrowia lipolytica ... [more ▼]

Strong constitutive promoter pTEF were used to overexpress genes encoding the two enzymes (L-cysteine gamma-ligase and glutathione synthase) which catalyze the synthesis of GSH in Yarrowia lipolytica. Several strains which can overexpress GSH1 or GSH2 or both of them have been successfully obtained. The newly constructed strains for glutathione overproduction show different compared to control. [less ▲]

Detailed reference viewed: 37 (6 ULiège)
Peer Reviewed
See detailCatabolic selectable marker eases genome editing in Yarrowia lipolytica
Vandermies, Marie ULiege; Denies, Olivia ULiege; Nicaud, Jean-Marc et al

Conference (2018, February 08)

Selectable markers are a central component of genome edition technologies. In the yeast Yarrowia lipolytica, these markers are traditionally based on antibiotic resistance (hygromycin B) or auxotrophy (e ... [more ▼]

Selectable markers are a central component of genome edition technologies. In the yeast Yarrowia lipolytica, these markers are traditionally based on antibiotic resistance (hygromycin B) or auxotrophy (e.g., leucine, uracil). However, the use of the former is impaired by a high level of spontaneous resistance, and the use of the latter by continuous complementation of the culture medium or restoration of prototrophy to the strains. As an alternative, genes related to the catabolism of carbon sources, or “catabolic selectable markers”, present the advantage of not being involved in essential metabolic pathways. The recently identified EYK1, encoding erythrulose kinase, can serve as an efficient catabolic selectable marker for genome editing in Y. lipolytica. Compared to auxotrophic markers such as URA3 and LEU2, EYK1 increases the growth rate of transformants on selective medium and the efficiency of genome edition. The utility of the marker EYK1 in a replicative vector was also demonstrated. Besides, the cloning-free strategy developed here simplifies the construction of disruption cassettes for genome editing in Y. lipolytica. [less ▲]

Detailed reference viewed: 121 (8 ULiège)
Peer Reviewed
See detailVersatile regulated promoter for Yarrowia lipolytica
Vandermies, Marie ULiege; Trassaert, Marion; Carly, Frédéric et al

Poster (2018, February 08)

The non-conventional yeast Yarrowia lipolytica is an emerging cell factory for the production of recombinant protein and metabolites. Still, there is a lack of versatile inducible promoters in this yeast ... [more ▼]

The non-conventional yeast Yarrowia lipolytica is an emerging cell factory for the production of recombinant protein and metabolites. Still, there is a lack of versatile inducible promoters in this yeast. In the present study, we isolated and characterised the promoter of the recently discovered gene EYK1 (encoding an erythrulose kinase). The promoter pEYK1 shows high induction levels in presence of erythritol and erythrulose, while its induction is impaired in presence of glycerol and glucose. Based on pEYK1 upstream activating sequence, we developed a series of promoters with enhanced induction levels. The induction levels of pEYK1 were also improved in strain eyk1Δ, allowing to use erythritol and erythrulose as free inducers. [less ▲]

Detailed reference viewed: 92 (5 ULiège)
Peer Reviewed
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 ▲]

Detailed reference viewed: 144 (11 ULiège)
Peer Reviewed
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 ▲]

Detailed reference viewed: 92 (7 ULiège)
Full Text
Peer Reviewed
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 ▲]

Detailed reference viewed: 38 (5 ULiège)
Full Text
Peer Reviewed
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 ▲]

Detailed reference viewed: 93 (10 ULiège)