Article (Scientific journals)
New inducible promoter for gene expression and synthetic biology in Yarrowia lipolytica
Trassaert, Marion; Vandermies, Marie; Carly, Frédéric et al.
2017In Microbial Cell Factories, 16 (141)
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Keywords :
Promoter; Regulation; Induction; Synthetic promoter; Erythritol; Erythrulose; Upstream activating sequence; Yarrowia lipolytica; Flow cytometry
Abstract :
[en] 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.
Disciplines :
Biotechnology
Author, co-author :
Trassaert, Marion 
Vandermies, Marie  ;  Université de Liège > Agronomie, Bio-ingénierie et Chimie (AgroBioChem) > Microbial, food and biobased technologies
Carly, Frédéric
Denies, Olivia ;  Université de Liège > Agronomie, Bio-ingénierie et Chimie (AgroBioChem) > Microbial, food and biobased technologies
Thomas, Stéphane
Fickers, Patrick ;  Université de Liège > Agronomie, Bio-ingénierie et Chimie (AgroBioChem) > Microbial, food and biobased technologies
Nicaud, Jean-Marc
 These authors have contributed equally to this work.
Language :
English
Title :
New inducible promoter for gene expression and synthetic biology in Yarrowia lipolytica
Publication date :
2017
Journal title :
Microbial Cell Factories
eISSN :
1475-2859
Publisher :
BioMed Central
Volume :
16
Issue :
141
Peer reviewed :
Peer Reviewed verified by ORBi
Funders :
FRIA - Fonds pour la Formation à la Recherche dans l'Industrie et dans l'Agriculture [BE]
Available on ORBi :
since 25 August 2017

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