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

Abstract : 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 (UAS1(EYK1)) and XPR2 (encoding extracellular protease, UAS1(XPR2)) 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.
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https://hal.inrae.fr/hal-02942667
Contributor : Christopher Lallemant <>
Submitted on : Friday, September 18, 2020 - 10:23:54 AM
Last modification on : Tuesday, June 15, 2021 - 2:56:38 PM

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Young-Kyoung Park, Marie Vandermies, Paul Soudier, Samuel Telek, Stéphane Thomas, et al.. Efficient expression vectors and host strain for the production of recombinant proteins by Yarrowia lipolytica in process conditions. Microbial Cell Factories, BioMed Central, 2019, 18 (1), ⟨10.1186/s12934-019-1218-6⟩. ⟨hal-02942667⟩

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