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Microbial synthesis of the plant natural product precursor p-coumaric acid with Corynebacterium glutamicum

BACKGROUND: Phenylpropanoids such as p-coumaric acid represent important precursors for the synthesis of a broad range of plant secondary metabolites including stilbenoids, flavonoids, and lignans, which are of pharmacological interest due to their health-promoting properties. Although extraction fr...

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Autores principales: Mutz, Mario, Kösters, Dominic, Wynands, Benedikt, Wierckx, Nick, Marienhagen, Jan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10576375/
https://www.ncbi.nlm.nih.gov/pubmed/37833813
http://dx.doi.org/10.1186/s12934-023-02222-y
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author Mutz, Mario
Kösters, Dominic
Wynands, Benedikt
Wierckx, Nick
Marienhagen, Jan
author_facet Mutz, Mario
Kösters, Dominic
Wynands, Benedikt
Wierckx, Nick
Marienhagen, Jan
author_sort Mutz, Mario
collection PubMed
description BACKGROUND: Phenylpropanoids such as p-coumaric acid represent important precursors for the synthesis of a broad range of plant secondary metabolites including stilbenoids, flavonoids, and lignans, which are of pharmacological interest due to their health-promoting properties. Although extraction from plant material or chemical synthesis is possible, microbial synthesis of p-coumaric acid from glucose has the advantage of being less expensive and more resource efficient. In this study, Corynebacterium glutamicum was engineered for the production of the plant polyphenol precursor p-coumaric acid from glucose. RESULTS: Heterologous expression of the tyrosine ammonia-lyase encoding gene from Flavobacterium johnsoniae enabled the conversion of endogenously provided tyrosine to p-coumaric acid. Product consumption was avoided by abolishing essential reactions of the phenylpropanoid degradation pathway. Accumulation of anthranilate as a major byproduct was eliminated by reducing the activity of anthranilate synthase through targeted mutagenesis to avoid tryptophan auxotrophy. Subsequently, the carbon flux into the shikimate pathway was increased, phenylalanine biosynthesis was reduced, and phosphoenolpyruvate availability was improved to boost p-coumaric acid accumulation. A maximum titer of 661 mg/L p-coumaric acid (4 mM) in defined mineral medium was reached. Finally, the production strain was utilized in co-cultivations with a C. glutamicum strain previously engineered for the conversion of p-coumaric acid into the polyphenol resveratrol. These co-cultivations enabled the synthesis of 31.2 mg/L (0.14 mM) resveratrol from glucose without any p-coumaric acid supplementation. CONCLUSIONS: The utilization of a heterologous tyrosine ammonia-lyase in combination with optimization of the shikimate pathway enabled the efficient production of p-coumaric acid with C. glutamicum. Reducing the carbon flux into the phenylalanine and tryptophan branches was the key to success along with the introduction of feedback-resistant enzyme variants. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-023-02222-y.
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spelling pubmed-105763752023-10-15 Microbial synthesis of the plant natural product precursor p-coumaric acid with Corynebacterium glutamicum Mutz, Mario Kösters, Dominic Wynands, Benedikt Wierckx, Nick Marienhagen, Jan Microb Cell Fact Research BACKGROUND: Phenylpropanoids such as p-coumaric acid represent important precursors for the synthesis of a broad range of plant secondary metabolites including stilbenoids, flavonoids, and lignans, which are of pharmacological interest due to their health-promoting properties. Although extraction from plant material or chemical synthesis is possible, microbial synthesis of p-coumaric acid from glucose has the advantage of being less expensive and more resource efficient. In this study, Corynebacterium glutamicum was engineered for the production of the plant polyphenol precursor p-coumaric acid from glucose. RESULTS: Heterologous expression of the tyrosine ammonia-lyase encoding gene from Flavobacterium johnsoniae enabled the conversion of endogenously provided tyrosine to p-coumaric acid. Product consumption was avoided by abolishing essential reactions of the phenylpropanoid degradation pathway. Accumulation of anthranilate as a major byproduct was eliminated by reducing the activity of anthranilate synthase through targeted mutagenesis to avoid tryptophan auxotrophy. Subsequently, the carbon flux into the shikimate pathway was increased, phenylalanine biosynthesis was reduced, and phosphoenolpyruvate availability was improved to boost p-coumaric acid accumulation. A maximum titer of 661 mg/L p-coumaric acid (4 mM) in defined mineral medium was reached. Finally, the production strain was utilized in co-cultivations with a C. glutamicum strain previously engineered for the conversion of p-coumaric acid into the polyphenol resveratrol. These co-cultivations enabled the synthesis of 31.2 mg/L (0.14 mM) resveratrol from glucose without any p-coumaric acid supplementation. CONCLUSIONS: The utilization of a heterologous tyrosine ammonia-lyase in combination with optimization of the shikimate pathway enabled the efficient production of p-coumaric acid with C. glutamicum. Reducing the carbon flux into the phenylalanine and tryptophan branches was the key to success along with the introduction of feedback-resistant enzyme variants. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-023-02222-y. BioMed Central 2023-10-13 /pmc/articles/PMC10576375/ /pubmed/37833813 http://dx.doi.org/10.1186/s12934-023-02222-y Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Mutz, Mario
Kösters, Dominic
Wynands, Benedikt
Wierckx, Nick
Marienhagen, Jan
Microbial synthesis of the plant natural product precursor p-coumaric acid with Corynebacterium glutamicum
title Microbial synthesis of the plant natural product precursor p-coumaric acid with Corynebacterium glutamicum
title_full Microbial synthesis of the plant natural product precursor p-coumaric acid with Corynebacterium glutamicum
title_fullStr Microbial synthesis of the plant natural product precursor p-coumaric acid with Corynebacterium glutamicum
title_full_unstemmed Microbial synthesis of the plant natural product precursor p-coumaric acid with Corynebacterium glutamicum
title_short Microbial synthesis of the plant natural product precursor p-coumaric acid with Corynebacterium glutamicum
title_sort microbial synthesis of the plant natural product precursor p-coumaric acid with corynebacterium glutamicum
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10576375/
https://www.ncbi.nlm.nih.gov/pubmed/37833813
http://dx.doi.org/10.1186/s12934-023-02222-y
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