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...
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ftdoajarticles:oai:doaj.org/article:efd896a9729c4daf989428e49745c7ad 2023-05-15T14:04:33+02:00 Efficient expression vectors and host strain for the production of recombinant proteins by Yarrowia lipolytica in process conditions Young-Kyoung Park Marie Vandermies Paul Soudier Samuel Telek Stéphane Thomas Jean-Marc Nicaud Patrick Fickers 2019-10-01T00:00:00Z https://doi.org/10.1186/s12934-019-1218-6 https://doaj.org/article/efd896a9729c4daf989428e49745c7ad EN eng BMC http://link.springer.com/article/10.1186/s12934-019-1218-6 https://doaj.org/toc/1475-2859 doi:10.1186/s12934-019-1218-6 1475-2859 https://doaj.org/article/efd896a9729c4daf989428e49745c7ad Microbial Cell Factories, Vol 18, Iss 1, Pp 1-12 (2019) Promoter Regulation Induction Synthetic promoter Erythritol Protein secretion Microbiology QR1-502 article 2019 ftdoajarticles https://doi.org/10.1186/s12934-019-1218-6 2022-12-31T15:46:14Z 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 (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. Article in Journal/Newspaper Antarc* Antarctica Directory of Open Access Journals: DOAJ Articles Microbial Cell Factories 18 1 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Promoter Regulation Induction Synthetic promoter Erythritol Protein secretion Microbiology QR1-502 |
spellingShingle |
Promoter Regulation Induction Synthetic promoter Erythritol Protein secretion Microbiology QR1-502 Young-Kyoung Park Marie Vandermies Paul Soudier Samuel Telek Stéphane Thomas Jean-Marc Nicaud Patrick Fickers Efficient expression vectors and host strain for the production of recombinant proteins by Yarrowia lipolytica in process conditions |
topic_facet |
Promoter Regulation Induction Synthetic promoter Erythritol Protein secretion Microbiology QR1-502 |
description |
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 (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. |
format |
Article in Journal/Newspaper |
author |
Young-Kyoung Park Marie Vandermies Paul Soudier Samuel Telek Stéphane Thomas Jean-Marc Nicaud Patrick Fickers |
author_facet |
Young-Kyoung Park Marie Vandermies Paul Soudier Samuel Telek Stéphane Thomas Jean-Marc Nicaud Patrick Fickers |
author_sort |
Young-Kyoung Park |
title |
Efficient expression vectors and host strain for the production of recombinant proteins by Yarrowia lipolytica in process conditions |
title_short |
Efficient expression vectors and host strain for the production of recombinant proteins by Yarrowia lipolytica in process conditions |
title_full |
Efficient expression vectors and host strain for the production of recombinant proteins by Yarrowia lipolytica in process conditions |
title_fullStr |
Efficient expression vectors and host strain for the production of recombinant proteins by Yarrowia lipolytica in process conditions |
title_full_unstemmed |
Efficient expression vectors and host strain for the production of recombinant proteins by Yarrowia lipolytica in process conditions |
title_sort |
efficient expression vectors and host strain for the production of recombinant proteins by yarrowia lipolytica in process conditions |
publisher |
BMC |
publishDate |
2019 |
url |
https://doi.org/10.1186/s12934-019-1218-6 https://doaj.org/article/efd896a9729c4daf989428e49745c7ad |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_source |
Microbial Cell Factories, Vol 18, Iss 1, Pp 1-12 (2019) |
op_relation |
http://link.springer.com/article/10.1186/s12934-019-1218-6 https://doaj.org/toc/1475-2859 doi:10.1186/s12934-019-1218-6 1475-2859 https://doaj.org/article/efd896a9729c4daf989428e49745c7ad |
op_doi |
https://doi.org/10.1186/s12934-019-1218-6 |
container_title |
Microbial Cell Factories |
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18 |
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1 |
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1766275711395430400 |