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Glucose limited feed strategy leads to increased production of fusicocca-2,10(14)-diene by Saccharomyces cerevisiae

Fusicocca-2,10(14)-diene (FCdiene) is a diterpene which is interesting as a precursor of the anti-cancer drug fusicoccin A and therefore for pharmaceutical applications. Production of FCdiene using a genetically modified Saccharomyces cerevisiae has been previously demonstrated with batch cultivatio...

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Autores principales: Halka, Lisa Marie, Nowacki, Christian, Kleinschmidt, Alica, Koenen, Kevin, Wichmann, Rolf
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6104463/
https://www.ncbi.nlm.nih.gov/pubmed/30136000
http://dx.doi.org/10.1186/s13568-018-0662-8
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author Halka, Lisa Marie
Nowacki, Christian
Kleinschmidt, Alica
Koenen, Kevin
Wichmann, Rolf
author_facet Halka, Lisa Marie
Nowacki, Christian
Kleinschmidt, Alica
Koenen, Kevin
Wichmann, Rolf
author_sort Halka, Lisa Marie
collection PubMed
description Fusicocca-2,10(14)-diene (FCdiene) is a diterpene which is interesting as a precursor of the anti-cancer drug fusicoccin A and therefore for pharmaceutical applications. Production of FCdiene using a genetically modified Saccharomyces cerevisiae has been previously demonstrated with batch cultivations with a product concentration up to 10 mg/L. However, it is widely known that fed-batch processes can significantly improve product titer in yeast fermentations. This study focuses on the establishment of fed-batch fermentation for FCdiene production because fed-batch cultivations using FeedBeads(®) indicated that limiting glucose supply could increase yields of biomass (1.07 g(CDW)/g(Glucose) instead of 0.20 g(CDW)/g(Glucose)) and FCdiene (21.54 mg(FCdiene)/g(Glucose) instead of 9.74 mg(FCdiene)/g(Glucose)) in shake flask scale and may have implications for larger scale processes. We implemented a new exponential glucose feed profile in a 1.8 L stirred tank reactor. This reduced overfeeding and the consequent, ethanol production. As a result improvements in cell concentrations up to 246% could be achieved and FCdiene yield increased up to 2.8X in the first 28 h. FCdiene concentration reached 161 mg/L and 320 mg/L at 44 h. Fed-batch and batch mode were combined to examine dynamics of bi-modal cultivation where a fed-batch phase was used for biomass production and a batch phase used for FCdiene production potentially supported by ethanol consumption as reported on production of betulinic acid. The present study highlights the potential of process development improvements which increase high-value heterologous diterpene yields from S. cerevisiae. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13568-018-0662-8) contains supplementary material, which is available to authorized users.
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spelling pubmed-61044632018-09-04 Glucose limited feed strategy leads to increased production of fusicocca-2,10(14)-diene by Saccharomyces cerevisiae Halka, Lisa Marie Nowacki, Christian Kleinschmidt, Alica Koenen, Kevin Wichmann, Rolf AMB Express Original Article Fusicocca-2,10(14)-diene (FCdiene) is a diterpene which is interesting as a precursor of the anti-cancer drug fusicoccin A and therefore for pharmaceutical applications. Production of FCdiene using a genetically modified Saccharomyces cerevisiae has been previously demonstrated with batch cultivations with a product concentration up to 10 mg/L. However, it is widely known that fed-batch processes can significantly improve product titer in yeast fermentations. This study focuses on the establishment of fed-batch fermentation for FCdiene production because fed-batch cultivations using FeedBeads(®) indicated that limiting glucose supply could increase yields of biomass (1.07 g(CDW)/g(Glucose) instead of 0.20 g(CDW)/g(Glucose)) and FCdiene (21.54 mg(FCdiene)/g(Glucose) instead of 9.74 mg(FCdiene)/g(Glucose)) in shake flask scale and may have implications for larger scale processes. We implemented a new exponential glucose feed profile in a 1.8 L stirred tank reactor. This reduced overfeeding and the consequent, ethanol production. As a result improvements in cell concentrations up to 246% could be achieved and FCdiene yield increased up to 2.8X in the first 28 h. FCdiene concentration reached 161 mg/L and 320 mg/L at 44 h. Fed-batch and batch mode were combined to examine dynamics of bi-modal cultivation where a fed-batch phase was used for biomass production and a batch phase used for FCdiene production potentially supported by ethanol consumption as reported on production of betulinic acid. The present study highlights the potential of process development improvements which increase high-value heterologous diterpene yields from S. cerevisiae. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13568-018-0662-8) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2018-08-22 /pmc/articles/PMC6104463/ /pubmed/30136000 http://dx.doi.org/10.1186/s13568-018-0662-8 Text en © The Author(s) 2018 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.
spellingShingle Original Article
Halka, Lisa Marie
Nowacki, Christian
Kleinschmidt, Alica
Koenen, Kevin
Wichmann, Rolf
Glucose limited feed strategy leads to increased production of fusicocca-2,10(14)-diene by Saccharomyces cerevisiae
title Glucose limited feed strategy leads to increased production of fusicocca-2,10(14)-diene by Saccharomyces cerevisiae
title_full Glucose limited feed strategy leads to increased production of fusicocca-2,10(14)-diene by Saccharomyces cerevisiae
title_fullStr Glucose limited feed strategy leads to increased production of fusicocca-2,10(14)-diene by Saccharomyces cerevisiae
title_full_unstemmed Glucose limited feed strategy leads to increased production of fusicocca-2,10(14)-diene by Saccharomyces cerevisiae
title_short Glucose limited feed strategy leads to increased production of fusicocca-2,10(14)-diene by Saccharomyces cerevisiae
title_sort glucose limited feed strategy leads to increased production of fusicocca-2,10(14)-diene by saccharomyces cerevisiae
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6104463/
https://www.ncbi.nlm.nih.gov/pubmed/30136000
http://dx.doi.org/10.1186/s13568-018-0662-8
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