Journal Article

New inducible promoter for gene expression and synthetic biology in Yarrowia lipolytica

Denies, Olivia, Vandermies, Marie, Nicaud, Jean-Marc, Thomas, Stéphane, Trassaert, Marion, Carly, Fréderic, Fickers, Patrick

2017

Overview

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 UAS1 EYK1 . New hybrid promoters containing tandem repeats of either UAS1 XPR2 or UAS1 EYK1 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 .

• Publisher: BioMed Central,BioMed Central Ltd,BMC
• Subject: Applied Microbiology / Base Sequence / Biotechnologie / Biotechnology / Chemistry / Chemistry and Materials Science / Enzymology / Erythritol / Erythrulose / Flow cytometry / Fungal Proteins - genetics / Gene Expression - drug effects / Genetic aspects / Genetic Engineering / Induction / Life Sciences / Microbial genetic engineering / Microbial Genetics and Genomics / Microbiology / Physiological aspects / Plasmids - genetics / Plasmids - metabolism / Production processes / Promoter / Promoter Regions, Genetic / Promoters (Genetics) / Recombinant proteins / Recombinant Proteins - biosynthesis / Recombinant Proteins - genetics / Regulation / Sciences du vivant / Sequence Alignment / Synthetic promoter / Tandem Repeat Sequences - genetics / Tetroses - pharmacology / Upstream activating sequence / Yarrowia - growth & development / Yarrowia - metabolism / Yarrowia lipolytica / Yeast fungi