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Rewiring a secondary metabolite pathway towards itaconic acid production in Aspergillus niger

BACKGROUND: The industrially relevant filamentous fungus Aspergillus niger is widely used in industry for its secretion capabilities of enzymes and organic acids. Biotechnologically produced organic acids promise to be an attractive alternative for the chemical industry to replace petrochemicals. It...

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Autores principales: Hossain, Abeer H., Li, An, Brickwedde, Anja, Wilms, Lars, Caspers, Martien, Overkamp, Karin, Punt, Peter J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4965889/
https://www.ncbi.nlm.nih.gov/pubmed/27469970
http://dx.doi.org/10.1186/s12934-016-0527-2
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author Hossain, Abeer H.
Li, An
Brickwedde, Anja
Wilms, Lars
Caspers, Martien
Overkamp, Karin
Punt, Peter J.
author_facet Hossain, Abeer H.
Li, An
Brickwedde, Anja
Wilms, Lars
Caspers, Martien
Overkamp, Karin
Punt, Peter J.
author_sort Hossain, Abeer H.
collection PubMed
description BACKGROUND: The industrially relevant filamentous fungus Aspergillus niger is widely used in industry for its secretion capabilities of enzymes and organic acids. Biotechnologically produced organic acids promise to be an attractive alternative for the chemical industry to replace petrochemicals. Itaconic acid (IA) has been identified as one of the top twelve building block chemicals which have high potential to be produced by biotechnological means. The IA biosynthesis cluster (cadA, mttA and mfsA) has been elucidated in its natural producer Aspergillus terreus and transferred to A. niger to enable IA production. Here we report the rewiring of a secondary metabolite pathway towards further improved IA production through the overexpression of a putative cytosolic citrate synthase citB in a A. niger strain carrying the IA biosynthesis cluster. RESULTS: We have previously shown that expression of cadA from A. terreus results in itaconic acid production in A. niger AB1.13, albeit at low levels. This low-level production is boosted fivefold by the overexpression of mttA and mfsA in itaconic acid producing AB1.13 CAD background strains. Controlled batch cultivations with AB1.13 CAD + MFS + MTT strains showed increased production of itaconic acid compared with AB1.13 CAD strain. Moreover, preliminary RNA-Seq analysis of an itaconic acid producing AB1.13 CAD strain has led to the identification of the putative cytosolic citrate synthase citB which was induced in an IA producing strain. We have overexpressed citB in a AB1.13 CAD + MFS + MTT strain and by doing so hypothesize to have targeted itaconic acid production to the cytosolic compartment. By overexpressing citB in AB1.13 CAD + MFS + MTT strains in controlled batch cultivations we have achieved highly increased titers of up to 26.2 g/L IA with a productivity of 0.35 g/L/h while no CA was produced. CONCLUSIONS: Expression of the IA biosynthesis cluster in Aspergillus niger AB1.13 strain enables IA production. Moreover, in the AB1.13 CAD strain IA production resulted in overexpression of a putative cytosolic citrate synthase citB. Upon overexpression of citB we have achieved titers of up to 26.2 g/L IA with a productivity of 0.35 g/L/h in controlled batch cultivations. By overexpressing citB we have also diminished side product formation and optimized the production pathway towards IA. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12934-016-0527-2) contains supplementary material, which is available to authorized users.
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spelling pubmed-49658892016-07-30 Rewiring a secondary metabolite pathway towards itaconic acid production in Aspergillus niger Hossain, Abeer H. Li, An Brickwedde, Anja Wilms, Lars Caspers, Martien Overkamp, Karin Punt, Peter J. Microb Cell Fact Research BACKGROUND: The industrially relevant filamentous fungus Aspergillus niger is widely used in industry for its secretion capabilities of enzymes and organic acids. Biotechnologically produced organic acids promise to be an attractive alternative for the chemical industry to replace petrochemicals. Itaconic acid (IA) has been identified as one of the top twelve building block chemicals which have high potential to be produced by biotechnological means. The IA biosynthesis cluster (cadA, mttA and mfsA) has been elucidated in its natural producer Aspergillus terreus and transferred to A. niger to enable IA production. Here we report the rewiring of a secondary metabolite pathway towards further improved IA production through the overexpression of a putative cytosolic citrate synthase citB in a A. niger strain carrying the IA biosynthesis cluster. RESULTS: We have previously shown that expression of cadA from A. terreus results in itaconic acid production in A. niger AB1.13, albeit at low levels. This low-level production is boosted fivefold by the overexpression of mttA and mfsA in itaconic acid producing AB1.13 CAD background strains. Controlled batch cultivations with AB1.13 CAD + MFS + MTT strains showed increased production of itaconic acid compared with AB1.13 CAD strain. Moreover, preliminary RNA-Seq analysis of an itaconic acid producing AB1.13 CAD strain has led to the identification of the putative cytosolic citrate synthase citB which was induced in an IA producing strain. We have overexpressed citB in a AB1.13 CAD + MFS + MTT strain and by doing so hypothesize to have targeted itaconic acid production to the cytosolic compartment. By overexpressing citB in AB1.13 CAD + MFS + MTT strains in controlled batch cultivations we have achieved highly increased titers of up to 26.2 g/L IA with a productivity of 0.35 g/L/h while no CA was produced. CONCLUSIONS: Expression of the IA biosynthesis cluster in Aspergillus niger AB1.13 strain enables IA production. Moreover, in the AB1.13 CAD strain IA production resulted in overexpression of a putative cytosolic citrate synthase citB. Upon overexpression of citB we have achieved titers of up to 26.2 g/L IA with a productivity of 0.35 g/L/h in controlled batch cultivations. By overexpressing citB we have also diminished side product formation and optimized the production pathway towards IA. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12934-016-0527-2) contains supplementary material, which is available to authorized users. BioMed Central 2016-07-28 /pmc/articles/PMC4965889/ /pubmed/27469970 http://dx.doi.org/10.1186/s12934-016-0527-2 Text en © The Author(s) 2016 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
Hossain, Abeer H.
Li, An
Brickwedde, Anja
Wilms, Lars
Caspers, Martien
Overkamp, Karin
Punt, Peter J.
Rewiring a secondary metabolite pathway towards itaconic acid production in Aspergillus niger
title Rewiring a secondary metabolite pathway towards itaconic acid production in Aspergillus niger
title_full Rewiring a secondary metabolite pathway towards itaconic acid production in Aspergillus niger
title_fullStr Rewiring a secondary metabolite pathway towards itaconic acid production in Aspergillus niger
title_full_unstemmed Rewiring a secondary metabolite pathway towards itaconic acid production in Aspergillus niger
title_short Rewiring a secondary metabolite pathway towards itaconic acid production in Aspergillus niger
title_sort rewiring a secondary metabolite pathway towards itaconic acid production in aspergillus niger
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4965889/
https://www.ncbi.nlm.nih.gov/pubmed/27469970
http://dx.doi.org/10.1186/s12934-016-0527-2
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