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Production of β-ionone by combined expression of carotenogenic and plant CCD1 genes in Saccharomyces cerevisiae

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 plants—like raspberries and roses—by the enzymatic cleavage of carotenes. Due to their pleasant aroma and flavour, apocaroteno...

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Autores principales: López, Javiera, Essus, Karen, Kim, Il-kwon, Pereira, Rui, Herzog, Jan, Siewers, Verena, Nielsen, Jens, Agosin, Eduardo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4464609/
https://www.ncbi.nlm.nih.gov/pubmed/26063466
http://dx.doi.org/10.1186/s12934-015-0273-x
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author López, Javiera
Essus, Karen
Kim, Il-kwon
Pereira, Rui
Herzog, Jan
Siewers, Verena
Nielsen, Jens
Agosin, Eduardo
author_facet López, Javiera
Essus, Karen
Kim, Il-kwon
Pereira, Rui
Herzog, Jan
Siewers, Verena
Nielsen, Jens
Agosin, Eduardo
author_sort López, Javiera
collection PubMed
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 plants—like raspberries and roses—by 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 β-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 β-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 β-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 β-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 β-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 gene—the highest β-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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12934-015-0273-x) contains supplementary material, which is available to authorized users.
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spelling pubmed-44646092015-06-14 Production of β-ionone by combined expression of carotenogenic and plant CCD1 genes in Saccharomyces cerevisiae López, Javiera Essus, Karen Kim, Il-kwon Pereira, Rui Herzog, Jan Siewers, Verena Nielsen, Jens Agosin, Eduardo Microb Cell Fact Research 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 plants—like raspberries and roses—by 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 β-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 β-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 β-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 β-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 β-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 gene—the highest β-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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12934-015-0273-x) contains supplementary material, which is available to authorized users. BioMed Central 2015-06-12 /pmc/articles/PMC4464609/ /pubmed/26063466 http://dx.doi.org/10.1186/s12934-015-0273-x Text en © López et al. 2015 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
López, Javiera
Essus, Karen
Kim, Il-kwon
Pereira, Rui
Herzog, Jan
Siewers, Verena
Nielsen, Jens
Agosin, Eduardo
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
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_short 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
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4464609/
https://www.ncbi.nlm.nih.gov/pubmed/26063466
http://dx.doi.org/10.1186/s12934-015-0273-x
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