Downregulation by organic nitrogen of AOX1 promoter used for controlled expression of foreign genes in the yeast Pichia pastoris
peer reviewed Pichia pastoris is a well-established cell factory for recombinant protein synthesis. Various optimization strategies of processes based on AOX1 promoter have been investigated, including methanol co-feeding with glycerol or sorbitol during the induction stage. Compared with carbon sou...
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John Wiley and Sons Ltd
2019
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Online Access: | https://orbi.uliege.be/handle/2268/257043 https://doi.org/10.1002/yea.3381 |
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ftorbi:oai:orbi.ulg.ac.be:2268/257043 2024-10-13T14:03:02+00:00 Downregulation by organic nitrogen of AOX1 promoter used for controlled expression of foreign genes in the yeast Pichia pastoris Velastegui, E. Theron, C. Berrios, J. Fickers, Patrick 2019 https://orbi.uliege.be/handle/2268/257043 https://doi.org/10.1002/yea.3381 unknown John Wiley and Sons Ltd urn:issn:0749-503X urn:issn:1097-0061 https://orbi.uliege.be/handle/2268/257043 info:hdl:2268/257043 doi:10.1002/yea.3381 info:pmid:30699241 restricted access http://purl.org/coar/access_right/c_16ec info:eu-repo/semantics/restrictedAccess Yeast, 36 (5), 297-304 (2019) AOX1 promoter regulation P. pastoris AOX1 gene Article Candida antarctica Komagataella pastoris Pichia Amino Acids Down-Regulation Fermentation Gene Expression Regulation Fungal Green Fluorescent Proteins Lipase Methanol Nitrogen Oxidoreductases Promoter Regions Genetic Recombinant Proteins Life sciences Biotechnology Sciences du vivant Biotechnologie journal article http://purl.org/coar/resource_type/c_6501 info:eu-repo/semantics/article peer reviewed 2019 ftorbi https://doi.org/10.1002/yea.3381 2024-09-27T07:01:36Z peer reviewed Pichia pastoris is a well-established cell factory for recombinant protein synthesis. Various optimization strategies of processes based on AOX1 promoter have been investigated, including methanol co-feeding with glycerol or sorbitol during the induction stage. Compared with carbon sources, comparatively little research has been devoted to the effects of nitrogen sources. Several reports have described the benefits of adding casamino acids (CA) to the recombinant protein production medium, however, without considering its effects at the gene expression level. Using enhanced green fluorescent protein as a reporter protein, monitored using flow cytometry, CA was shown to downregulate AOX1 promoter induction. Despite higher growth rates, cultures containing CA exhibited slower transition to the induced state, whereas metabolite analysis revealed that methanol consumption was reduced in the presence of CA compared with its absence. The repressive effect of CA was further confirmed by analysing the synthesis of extracellular recombinant Candida antarctica lipase under control of the AOX1 promoter. These findings highlight nitrogen source selection as an important consideration for AOX1-based protein production. © 2019 John Wiley & Sons, Ltd. Article in Journal/Newspaper Antarc* Antarctica University of Liège: ORBi (Open Repository and Bibliography) Yeast 36 5 297 304 |
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University of Liège: ORBi (Open Repository and Bibliography) |
op_collection_id |
ftorbi |
language |
unknown |
topic |
AOX1 promoter regulation P. pastoris AOX1 gene Article Candida antarctica Komagataella pastoris Pichia Amino Acids Down-Regulation Fermentation Gene Expression Regulation Fungal Green Fluorescent Proteins Lipase Methanol Nitrogen Oxidoreductases Promoter Regions Genetic Recombinant Proteins Life sciences Biotechnology Sciences du vivant Biotechnologie |
spellingShingle |
AOX1 promoter regulation P. pastoris AOX1 gene Article Candida antarctica Komagataella pastoris Pichia Amino Acids Down-Regulation Fermentation Gene Expression Regulation Fungal Green Fluorescent Proteins Lipase Methanol Nitrogen Oxidoreductases Promoter Regions Genetic Recombinant Proteins Life sciences Biotechnology Sciences du vivant Biotechnologie Velastegui, E. Theron, C. Berrios, J. Fickers, Patrick Downregulation by organic nitrogen of AOX1 promoter used for controlled expression of foreign genes in the yeast Pichia pastoris |
topic_facet |
AOX1 promoter regulation P. pastoris AOX1 gene Article Candida antarctica Komagataella pastoris Pichia Amino Acids Down-Regulation Fermentation Gene Expression Regulation Fungal Green Fluorescent Proteins Lipase Methanol Nitrogen Oxidoreductases Promoter Regions Genetic Recombinant Proteins Life sciences Biotechnology Sciences du vivant Biotechnologie |
description |
peer reviewed Pichia pastoris is a well-established cell factory for recombinant protein synthesis. Various optimization strategies of processes based on AOX1 promoter have been investigated, including methanol co-feeding with glycerol or sorbitol during the induction stage. Compared with carbon sources, comparatively little research has been devoted to the effects of nitrogen sources. Several reports have described the benefits of adding casamino acids (CA) to the recombinant protein production medium, however, without considering its effects at the gene expression level. Using enhanced green fluorescent protein as a reporter protein, monitored using flow cytometry, CA was shown to downregulate AOX1 promoter induction. Despite higher growth rates, cultures containing CA exhibited slower transition to the induced state, whereas metabolite analysis revealed that methanol consumption was reduced in the presence of CA compared with its absence. The repressive effect of CA was further confirmed by analysing the synthesis of extracellular recombinant Candida antarctica lipase under control of the AOX1 promoter. These findings highlight nitrogen source selection as an important consideration for AOX1-based protein production. © 2019 John Wiley & Sons, Ltd. |
format |
Article in Journal/Newspaper |
author |
Velastegui, E. Theron, C. Berrios, J. Fickers, Patrick |
author_facet |
Velastegui, E. Theron, C. Berrios, J. Fickers, Patrick |
author_sort |
Velastegui, E. |
title |
Downregulation by organic nitrogen of AOX1 promoter used for controlled expression of foreign genes in the yeast Pichia pastoris |
title_short |
Downregulation by organic nitrogen of AOX1 promoter used for controlled expression of foreign genes in the yeast Pichia pastoris |
title_full |
Downregulation by organic nitrogen of AOX1 promoter used for controlled expression of foreign genes in the yeast Pichia pastoris |
title_fullStr |
Downregulation by organic nitrogen of AOX1 promoter used for controlled expression of foreign genes in the yeast Pichia pastoris |
title_full_unstemmed |
Downregulation by organic nitrogen of AOX1 promoter used for controlled expression of foreign genes in the yeast Pichia pastoris |
title_sort |
downregulation by organic nitrogen of aox1 promoter used for controlled expression of foreign genes in the yeast pichia pastoris |
publisher |
John Wiley and Sons Ltd |
publishDate |
2019 |
url |
https://orbi.uliege.be/handle/2268/257043 https://doi.org/10.1002/yea.3381 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_source |
Yeast, 36 (5), 297-304 (2019) |
op_relation |
urn:issn:0749-503X urn:issn:1097-0061 https://orbi.uliege.be/handle/2268/257043 info:hdl:2268/257043 doi:10.1002/yea.3381 info:pmid:30699241 |
op_rights |
restricted access http://purl.org/coar/access_right/c_16ec info:eu-repo/semantics/restrictedAccess |
op_doi |
https://doi.org/10.1002/yea.3381 |
container_title |
Yeast |
container_volume |
36 |
container_issue |
5 |
container_start_page |
297 |
op_container_end_page |
304 |
_version_ |
1812819462778257408 |