Efficient expression vectors and host strain for the production of recombinant proteins by Yarrowia lipolytica in process conditions

Park, Y.-K.; Vandermies, Marie; Soudier, P.; Telek, Samuel; Thomas, Stéphane; Nicaud, J.-M.; Fickers, Patrick

doi:10.1186/s12934-019-1218-6

Article (Scientific journals)

Efficient expression vectors and host strain for the production of recombinant proteins by Yarrowia lipolytica in process conditions

Park, Y.-K.; Vandermies, Marie; Soudier, P. et al.

2019 • In Microbial Cell Factories, 18 (1)

Peer Reviewed verified by ORBi

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https://hdl.handle.net/2268/257034

DOI
10.1186/s12934-019-1218-6

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31601223

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Keywords :

CalB; Erythritol; Induction; Promoter; Protein secretion; Regulation; Synthetic promoter; Upstream activating sequence; Yarrowia lipolytica; Article; CalB gene; EYD1 gene; EYK1 gene; XPR2 gene; Yarrowia; Fungal Proteins; Gene Expression; Lipase; Microorganisms, Genetically-Modified; Promoter Regions, Genetic; Recombinant Proteins

Abstract :

[en] Background: The oleaginous yeast Yarrowia lipolytica is increasingly used as an alternative cell factory for the production of recombinant proteins. Recently, regulated promoters from genes EYK1 and EYD1, encoding an erythrulose kinase and an erythritol dehydrogenase, respectively, have been identified and characterized in this yeast. Hybrid promoters up-regulated by polyols such as erythritol and erythrulose have been developed based on tandem copies of upstream activating sequences from EYK1 (UAS1EYK1) and XPR2 (encoding extracellular protease, UAS1XPR2) promoters. Results: The strength of native (pEYD1) and engineered promoters (pEYK1-3AB and pHU8EYK) was compared using the extracellular lipase CalB from Candida antarctica as a model protein and a novel dedicated host strain. This latter is engineered in polyol metabolism and allows targeted chromosomal integration. In process conditions, engineered promoters pEYK1-3AB and pHU8EYK yielded 2.8 and 2.5-fold higher protein productivity, respectively, as compared to the reference pTEF promoter. We also demonstrated the possibility of multicopy integration in the newly developed host strain. In batch bioreactor, the CalB multi-copy strain RIY406 led to a 1.6 fold increased lipase productivity (45,125 U mL-1) within 24 h as compared to the mono-copy strain. Conclusions: The expression system described herein appears promising for recombinant extracellular protein production in Y. lipolytica. © 2019 The Author(s).

Disciplines :

Biotechnology

Author, co-author :

Park, Y.-K.; Micalis Institute, INRA, AgroParisTech, Université Paris-Saclay, Jouy-en-Josas, 78350, France

Vandermies, Marie ; Université de Liège - ULiège > Terra

Soudier, P.; Micalis Institute, INRA, AgroParisTech, Université Paris-Saclay, Jouy-en-Josas, 78350, France

Telek, Samuel ; Université de Liège - ULiège > Département GxABT > Microbial, food and biobased technologies

Thomas, Stéphane

Nicaud, J.-M.; Micalis Institute, INRA, AgroParisTech, Université Paris-Saclay, Jouy-en-Josas, 78350, France, Micalis Institute, UMR1319, Team BIMLip: Integrative Metabolism of Microbial Lipids, INRA-AgroParisTech, Domaine de Vilvert, Jouy-en-Josas, 78352, France

Fickers, Patrick ; Université de Liège - ULiège > Département GxABT > Microbial, food and biobased technologies

Language :

English

Title :

Efficient expression vectors and host strain for the production of recombinant proteins by Yarrowia lipolytica in process conditions

Publication date :

2019

Journal title :

Microbial Cell Factories

eISSN :

1475-2859

Publisher :

BioMed Central Ltd.

Volume :

Issue :

Peer reviewed :

Peer Reviewed verified by ORBi

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since 18 February 2021

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