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

• Published in: Microbial Cell Factories
• Main Authors: Park, Young-Kyoung, Vandermies, Marie, Soudier, Paul, Telek, Samuel, Thomas, Stéphane, Nicaud, Jean-Marc, Fickers, Patrick
• Other Authors: MICrobiologie de l'ALImentation au Service de la Santé (MICALIS), Institut National de la Recherche Agronomique (INRA)-AgroParisTech, Université Paris-Saclay, Microbial Processes and Interactions, ERRA Teaching and Research Centre, Université de Liège - Gembloux-Université de Liège - Gembloux, INRA Biocatalysts Fonds de la Recherche Scientifique - FNRSKwanjeong Educational Foundation (KEF)
• Format: Article in Journal/Newspaper
• Language: English
• Published: HAL CCSD 2019
• Subjects: Promoter; Regulation; Induction; Synthetic promoter; Erythritol; Protein secretion; Upstream activating sequence; Yarrowia lipolytica; CalB; [SDV.BIO]Life Sciences [q-bio]/Biotechnology; Antarc*; Antarctica
• Online Access: https://hal.inrae.fr/hal-02942667; https://doi.org/10.1186/s12934-019-1218-6

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.