Production of β‑ionone by combined expression of carotenogenic and plant CCD1 genes in Saccharomyces cerevisiae
Autor(a) principal: | |
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Data de Publicação: | 2015 |
Outros Autores: | , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/1822/35812 |
Resumo: | Background Apocarotenoids, like the C13-norisoprenoids, are natural compounds that contribute to the flavor and/or aroma of flowers and foods. They are produced in aromatic plantslike raspberries and rosesby the enzymatic cleavage of carotenes. Due to their pleasant aroma and flavour, apocarotenoids have high commercial value for the cosmetic and food industry, but currently their production is mainly assured by chemical synthesis. In the present study, a Saccharomyces cerevisiae strain that synthesizes the apocarotenoid -ionone was constructed by combining integrative vectors and high copy number episomal vectors, in an engineered strain that accumulates FPP. Results Integration of an extra copy of the geranylgeranyl diphosphate synthase gene (BTS1), together with the carotenogenic genes crtYB and crtI from the ascomycete Xanthophyllomyces dendrorhous, resulted in carotenoid producing cells. The additional integration of the carotenoid cleavage dioxygenase gene from the plant Petunia hybrida (PhCCD1) let to the production of low amounts of -ionone (0.073 ± 0.01 mg/g DCW) and changed the color of the strain from orange to yellow. The expression of the crtYB gene from a high copy number plasmid in this former strain increased -ionone concentration fivefold (0.34 ± 0.06 mg/g DCW). Additionally, the episomal expression of crtYB together with the PhCCD1 gene in the same vector resulted in a final 8.5-fold increase of -ionone concentration (0.63 ± 0.02 mg/g DCW). Batch fermentations with this strain resulted in a final specific concentration of 1 mg/g DCW at 50 h, which represents a 15-fold increase. Conclusions An efficient -ionone producing yeast platform was constructed by combining integrative and episomal constructs. By combined expression of the genes BTS1, the carotenogenic crtYB, crtI genes and the plant PhCCD1 genethe highest -ionone concentration reported to date by a cell factory was achieved. This microbial cell factory represents a starting point for flavor production by a sustainable and efficient process that could replace current methods. |
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Production of β‑ionone by combined expression of carotenogenic and plant CCD1 genes in Saccharomyces cerevisiaeMetabolic engineeringCarotenoidsApocarotenoidsSaccharomyces cerevisiaeCiências Naturais::Ciências BiológicasEngenharia e Tecnologia::Biotecnologia IndustrialScience & TechnologyBackground Apocarotenoids, like the C13-norisoprenoids, are natural compounds that contribute to the flavor and/or aroma of flowers and foods. They are produced in aromatic plantslike raspberries and rosesby the enzymatic cleavage of carotenes. Due to their pleasant aroma and flavour, apocarotenoids have high commercial value for the cosmetic and food industry, but currently their production is mainly assured by chemical synthesis. In the present study, a Saccharomyces cerevisiae strain that synthesizes the apocarotenoid -ionone was constructed by combining integrative vectors and high copy number episomal vectors, in an engineered strain that accumulates FPP. Results Integration of an extra copy of the geranylgeranyl diphosphate synthase gene (BTS1), together with the carotenogenic genes crtYB and crtI from the ascomycete Xanthophyllomyces dendrorhous, resulted in carotenoid producing cells. The additional integration of the carotenoid cleavage dioxygenase gene from the plant Petunia hybrida (PhCCD1) let to the production of low amounts of -ionone (0.073 ± 0.01 mg/g DCW) and changed the color of the strain from orange to yellow. The expression of the crtYB gene from a high copy number plasmid in this former strain increased -ionone concentration fivefold (0.34 ± 0.06 mg/g DCW). Additionally, the episomal expression of crtYB together with the PhCCD1 gene in the same vector resulted in a final 8.5-fold increase of -ionone concentration (0.63 ± 0.02 mg/g DCW). Batch fermentations with this strain resulted in a final specific concentration of 1 mg/g DCW at 50 h, which represents a 15-fold increase. Conclusions An efficient -ionone producing yeast platform was constructed by combining integrative and episomal constructs. By combined expression of the genes BTS1, the carotenogenic crtYB, crtI genes and the plant PhCCD1 genethe highest -ionone concentration reported to date by a cell factory was achieved. This microbial cell factory represents a starting point for flavor production by a sustainable and efficient process that could replace current methods.This work was funded by grants COPEC-UC 6C-063 and FONDECYT No 1130822, and the Novo Nordisk Foundation.BioMed Central (BMC)Universidade do MinhoLópez, JavieraEssus, KarenKim, Il-kwonPereira, Rui Miguel Pinheiro SilvaHerzog, JanSiewers, VerenaNielsen, JensAgosin, Eduardo2015-062015-06-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/1822/35812engLópez, Javiera; Essus, Karen; Kim, Il-kwon; ; Herzog, Jan; Siewers, Verena; Nielsen, Jens; Agosin, Eduardo, Production of -ionone by combined expression of carotenogenic and plant CCD1 genes in Saccharomyces cerevisiae. Microbial Cell Factories, 14(84), 1-13, 20151475-28591475-285910.1186/s12934-015-0273-x26063466http://www.microbialcellfactories.com/content/14/1/84info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-21T12:21:26Zoai:repositorium.sdum.uminho.pt:1822/35812Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T19:14:45.011546Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Production of β‑ionone by combined expression of carotenogenic and plant CCD1 genes in Saccharomyces cerevisiae |
title |
Production of β‑ionone by combined expression of carotenogenic and plant CCD1 genes in Saccharomyces cerevisiae |
spellingShingle |
Production of β‑ionone by combined expression of carotenogenic and plant CCD1 genes in Saccharomyces cerevisiae López, Javiera Metabolic engineering Carotenoids Apocarotenoids Saccharomyces cerevisiae Ciências Naturais::Ciências Biológicas Engenharia e Tecnologia::Biotecnologia Industrial Science & Technology |
title_short |
Production of β‑ionone by combined expression of carotenogenic and plant CCD1 genes in Saccharomyces cerevisiae |
title_full |
Production of β‑ionone by combined expression of carotenogenic and plant CCD1 genes in Saccharomyces cerevisiae |
title_fullStr |
Production of β‑ionone by combined expression of carotenogenic and plant CCD1 genes in Saccharomyces cerevisiae |
title_full_unstemmed |
Production of β‑ionone by combined expression of carotenogenic and plant CCD1 genes in Saccharomyces cerevisiae |
title_sort |
Production of β‑ionone by combined expression of carotenogenic and plant CCD1 genes in Saccharomyces cerevisiae |
author |
López, Javiera |
author_facet |
López, Javiera Essus, Karen Kim, Il-kwon Pereira, Rui Miguel Pinheiro Silva Herzog, Jan Siewers, Verena Nielsen, Jens Agosin, Eduardo |
author_role |
author |
author2 |
Essus, Karen Kim, Il-kwon Pereira, Rui Miguel Pinheiro Silva Herzog, Jan Siewers, Verena Nielsen, Jens Agosin, Eduardo |
author2_role |
author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade do Minho |
dc.contributor.author.fl_str_mv |
López, Javiera Essus, Karen Kim, Il-kwon Pereira, Rui Miguel Pinheiro Silva Herzog, Jan Siewers, Verena Nielsen, Jens Agosin, Eduardo |
dc.subject.por.fl_str_mv |
Metabolic engineering Carotenoids Apocarotenoids Saccharomyces cerevisiae Ciências Naturais::Ciências Biológicas Engenharia e Tecnologia::Biotecnologia Industrial Science & Technology |
topic |
Metabolic engineering Carotenoids Apocarotenoids Saccharomyces cerevisiae Ciências Naturais::Ciências Biológicas Engenharia e Tecnologia::Biotecnologia Industrial Science & Technology |
description |
Background Apocarotenoids, like the C13-norisoprenoids, are natural compounds that contribute to the flavor and/or aroma of flowers and foods. They are produced in aromatic plantslike raspberries and rosesby the enzymatic cleavage of carotenes. Due to their pleasant aroma and flavour, apocarotenoids have high commercial value for the cosmetic and food industry, but currently their production is mainly assured by chemical synthesis. In the present study, a Saccharomyces cerevisiae strain that synthesizes the apocarotenoid -ionone was constructed by combining integrative vectors and high copy number episomal vectors, in an engineered strain that accumulates FPP. Results Integration of an extra copy of the geranylgeranyl diphosphate synthase gene (BTS1), together with the carotenogenic genes crtYB and crtI from the ascomycete Xanthophyllomyces dendrorhous, resulted in carotenoid producing cells. The additional integration of the carotenoid cleavage dioxygenase gene from the plant Petunia hybrida (PhCCD1) let to the production of low amounts of -ionone (0.073 ± 0.01 mg/g DCW) and changed the color of the strain from orange to yellow. The expression of the crtYB gene from a high copy number plasmid in this former strain increased -ionone concentration fivefold (0.34 ± 0.06 mg/g DCW). Additionally, the episomal expression of crtYB together with the PhCCD1 gene in the same vector resulted in a final 8.5-fold increase of -ionone concentration (0.63 ± 0.02 mg/g DCW). Batch fermentations with this strain resulted in a final specific concentration of 1 mg/g DCW at 50 h, which represents a 15-fold increase. Conclusions An efficient -ionone producing yeast platform was constructed by combining integrative and episomal constructs. By combined expression of the genes BTS1, the carotenogenic crtYB, crtI genes and the plant PhCCD1 genethe highest -ionone concentration reported to date by a cell factory was achieved. This microbial cell factory represents a starting point for flavor production by a sustainable and efficient process that could replace current methods. |
publishDate |
2015 |
dc.date.none.fl_str_mv |
2015-06 2015-06-01T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/1822/35812 |
url |
http://hdl.handle.net/1822/35812 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
López, Javiera; Essus, Karen; Kim, Il-kwon; ; Herzog, Jan; Siewers, Verena; Nielsen, Jens; Agosin, Eduardo, Production of -ionone by combined expression of carotenogenic and plant CCD1 genes in Saccharomyces cerevisiae. Microbial Cell Factories, 14(84), 1-13, 2015 1475-2859 1475-2859 10.1186/s12934-015-0273-x 26063466 http://www.microbialcellfactories.com/content/14/1/84 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
BioMed Central (BMC) |
publisher.none.fl_str_mv |
BioMed Central (BMC) |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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RCAAP |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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1799132590591967232 |