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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ftdatacite:10.6084/m9.figshare.c.4694783 2023-05-15T14:04:28+02:00 Efficient expression vectors and host strain for the production of recombinant proteins by Yarrowia lipolytica in process conditions Young-Kyoung Park Vandermies, Marie Soudier, Paul Telek, Samuel Thomas, Stéphane Jean-Marc Nicaud Fickers, Patrick 2019 https://dx.doi.org/10.6084/m9.figshare.c.4694783 https://springernature.figshare.com/collections/Efficient_expression_vectors_and_host_strain_for_the_production_of_recombinant_proteins_by_Yarrowia_lipolytica_in_process_conditions/4694783 unknown figshare https://dx.doi.org/10.1186/s12934-019-1218-6 CC BY 4.0 https://creativecommons.org/licenses/by/4.0 CC-BY Biochemistry Space Science Microbiology FOS Biological sciences Genetics Molecular Biology Biotechnology 69999 Biological Sciences not elsewhere classified Developmental Biology Collection article 2019 ftdatacite https://doi.org/10.6084/m9.figshare.c.4694783 https://doi.org/10.1186/s12934-019-1218-6 2021-11-05T12:55:41Z 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 DataCite Metadata Store (German National Library of Science and Technology) |
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Open Polar |
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DataCite Metadata Store (German National Library of Science and Technology) |
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ftdatacite |
language |
unknown |
topic |
Biochemistry Space Science Microbiology FOS Biological sciences Genetics Molecular Biology Biotechnology 69999 Biological Sciences not elsewhere classified Developmental Biology |
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Biochemistry Space Science Microbiology FOS Biological sciences Genetics Molecular Biology Biotechnology 69999 Biological Sciences not elsewhere classified Developmental Biology Young-Kyoung Park Vandermies, Marie Soudier, Paul Telek, Samuel Thomas, Stéphane Jean-Marc Nicaud Fickers, Patrick Efficient expression vectors and host strain for the production of recombinant proteins by Yarrowia lipolytica in process conditions |
topic_facet |
Biochemistry Space Science Microbiology FOS Biological sciences Genetics Molecular Biology Biotechnology 69999 Biological Sciences not elsewhere classified Developmental Biology |
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 Vandermies, Marie Soudier, Paul Telek, Samuel Thomas, Stéphane Jean-Marc Nicaud Fickers, Patrick |
author_facet |
Young-Kyoung Park Vandermies, Marie Soudier, Paul Telek, Samuel Thomas, Stéphane Jean-Marc Nicaud Fickers, Patrick |
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 |
figshare |
publishDate |
2019 |
url |
https://dx.doi.org/10.6084/m9.figshare.c.4694783 https://springernature.figshare.com/collections/Efficient_expression_vectors_and_host_strain_for_the_production_of_recombinant_proteins_by_Yarrowia_lipolytica_in_process_conditions/4694783 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_relation |
https://dx.doi.org/10.1186/s12934-019-1218-6 |
op_rights |
CC BY 4.0 https://creativecommons.org/licenses/by/4.0 |
op_rightsnorm |
CC-BY |
op_doi |
https://doi.org/10.6084/m9.figshare.c.4694783 https://doi.org/10.1186/s12934-019-1218-6 |
_version_ |
1766275574770171904 |