Selective production of the itaconic acid-derived compounds 2-hydroxyparaconic and itatartaric acid
There is a strong interest in itaconic acid in the medical and pharmaceutical sectors, both as an anti-bacterial compound and as an immunoregulator in mammalian macrophages. Fungal hosts also produce itaconic acid, and in addition they can produce two derivatives 2-hydroxyparaconic and itatartaric a...
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Elsevier
2024-12-01
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| Series: | Metabolic Engineering Communications |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S221403012400021X |
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| author | Philipp Ernst Felicia Zlati Larissa Kever Astrid Wirtz Rainer Goldbaum Jörg Pietruszka Benedikt Wynands Julia Frunzke Nick Wierckx |
| author_facet | Philipp Ernst Felicia Zlati Larissa Kever Astrid Wirtz Rainer Goldbaum Jörg Pietruszka Benedikt Wynands Julia Frunzke Nick Wierckx |
| author_sort | Philipp Ernst |
| collection | DOAJ |
| description | There is a strong interest in itaconic acid in the medical and pharmaceutical sectors, both as an anti-bacterial compound and as an immunoregulator in mammalian macrophages. Fungal hosts also produce itaconic acid, and in addition they can produce two derivatives 2-hydroxyparaconic and itatartaric acid. Not much is known about these two derivatives, while their structural analogy to itaconate could open up several applications. In this study, we report the production of these two itaconate-derived compounds. By overexpressing the itaconate P450 monooxygenase Cyp3 in a previously engineered itaconate-overproducing Ustilago cynodontis strain, itaconate was converted to its lactone 2-hydroxyparaconate. The second product itatartarate is most likely the result of the subsequent lactone hydrolysis. A major challenge in the production of 2-hydroxyparaconate and itatartarate is their co-production with itaconate, leading to difficulties in their purification. Achieving high derivatives specificity was therefore the paramount objective. Different strategies were evaluated including process parameters such as substrate and pH, as well as strain engineering focusing on Cyp3 expression and product export. 2-hydroxyparaconate and itatartarate were successfully produced from glucose and glycerol, with the latter resulting in a higher derivatives specificity due to an overall slower metabolism on this non-preferred carbon source. The derivatives specificity could be further increased by metabolic engineering approaches including the exchange of the native itaconate transporter Itp1 with the Aspergillus terreus itaconate transporter MfsA. Both 2-hydroxyparaconate and itatartarate were recovered from fermentation supernatants following a pre-existing protocol. 2-hydroxyparaconate was recovered first through a process of evaporation, lactonization, and extraction with ethyl acetate. Subsequently, itatartarate could be obtained in the form of its sodium salt by saponification of the purified 2-hydroxyparaconate. Finally, several analytical methods were used to characterize the resulting products and their structures were confirmed by nuclear magnetic resonance spectroscopy. This work provides a promising foundation for obtaining 2-hydroxyparaconate and itatartarate in high purity and quantity. This will allow to unravel the full spectrum of potential applications of these novel compounds. |
| format | Article |
| id | doaj-art-5c1c7d78ef6a43f3867c96e6309788c4 |
| institution | Kabale University |
| issn | 2214-0301 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Elsevier |
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| series | Metabolic Engineering Communications |
| spelling | doaj-art-5c1c7d78ef6a43f3867c96e6309788c42024-12-17T04:59:38ZengElsevierMetabolic Engineering Communications2214-03012024-12-0119e00252Selective production of the itaconic acid-derived compounds 2-hydroxyparaconic and itatartaric acidPhilipp Ernst0Felicia Zlati1Larissa Kever2Astrid Wirtz3Rainer Goldbaum4Jörg Pietruszka5Benedikt Wynands6Julia Frunzke7Nick Wierckx8Institute of Bio- and Geosciences IBG-1: Biotechnology, Forschungszentrum Jülich GmbH, Wilhelm-Johnen-Straße, 52428 Jülich, GermanyInstitute of Bio- and Geosciences IBG-1: Biotechnology, Forschungszentrum Jülich GmbH, Wilhelm-Johnen-Straße, 52428 Jülich, GermanyInstitute of Bio- and Geosciences IBG-1: Biotechnology, Forschungszentrum Jülich GmbH, Wilhelm-Johnen-Straße, 52428 Jülich, GermanyInstitute of Bio- and Geosciences IBG-1: Biotechnology, Forschungszentrum Jülich GmbH, Wilhelm-Johnen-Straße, 52428 Jülich, GermanyInstitute of Bioorganic Chemistry, Heinrich-Heine University Düsseldorf in Forschungszentrum Jülich GmbH, Wilhelm-Johnen-Straße, 52428, Jülich, GermanyInstitute of Bio- and Geosciences IBG-1: Biotechnology, Forschungszentrum Jülich GmbH, Wilhelm-Johnen-Straße, 52428 Jülich, Germany; Institute of Bioorganic Chemistry, Heinrich-Heine University Düsseldorf in Forschungszentrum Jülich GmbH, Wilhelm-Johnen-Straße, 52428, Jülich, GermanyInstitute of Bio- and Geosciences IBG-1: Biotechnology, Forschungszentrum Jülich GmbH, Wilhelm-Johnen-Straße, 52428 Jülich, GermanyInstitute of Bio- and Geosciences IBG-1: Biotechnology, Forschungszentrum Jülich GmbH, Wilhelm-Johnen-Straße, 52428 Jülich, GermanyInstitute of Bio- and Geosciences IBG-1: Biotechnology, Forschungszentrum Jülich GmbH, Wilhelm-Johnen-Straße, 52428 Jülich, Germany; Corresponding author.There is a strong interest in itaconic acid in the medical and pharmaceutical sectors, both as an anti-bacterial compound and as an immunoregulator in mammalian macrophages. Fungal hosts also produce itaconic acid, and in addition they can produce two derivatives 2-hydroxyparaconic and itatartaric acid. Not much is known about these two derivatives, while their structural analogy to itaconate could open up several applications. In this study, we report the production of these two itaconate-derived compounds. By overexpressing the itaconate P450 monooxygenase Cyp3 in a previously engineered itaconate-overproducing Ustilago cynodontis strain, itaconate was converted to its lactone 2-hydroxyparaconate. The second product itatartarate is most likely the result of the subsequent lactone hydrolysis. A major challenge in the production of 2-hydroxyparaconate and itatartarate is their co-production with itaconate, leading to difficulties in their purification. Achieving high derivatives specificity was therefore the paramount objective. Different strategies were evaluated including process parameters such as substrate and pH, as well as strain engineering focusing on Cyp3 expression and product export. 2-hydroxyparaconate and itatartarate were successfully produced from glucose and glycerol, with the latter resulting in a higher derivatives specificity due to an overall slower metabolism on this non-preferred carbon source. The derivatives specificity could be further increased by metabolic engineering approaches including the exchange of the native itaconate transporter Itp1 with the Aspergillus terreus itaconate transporter MfsA. Both 2-hydroxyparaconate and itatartarate were recovered from fermentation supernatants following a pre-existing protocol. 2-hydroxyparaconate was recovered first through a process of evaporation, lactonization, and extraction with ethyl acetate. Subsequently, itatartarate could be obtained in the form of its sodium salt by saponification of the purified 2-hydroxyparaconate. Finally, several analytical methods were used to characterize the resulting products and their structures were confirmed by nuclear magnetic resonance spectroscopy. This work provides a promising foundation for obtaining 2-hydroxyparaconate and itatartarate in high purity and quantity. This will allow to unravel the full spectrum of potential applications of these novel compounds.http://www.sciencedirect.com/science/article/pii/S221403012400021XUstilago cynodontis2-Hydroxyparaconic and itatartaric acidDerivatives specificityMetabolic engineeringFermentationDownstream processing |
| spellingShingle | Philipp Ernst Felicia Zlati Larissa Kever Astrid Wirtz Rainer Goldbaum Jörg Pietruszka Benedikt Wynands Julia Frunzke Nick Wierckx Selective production of the itaconic acid-derived compounds 2-hydroxyparaconic and itatartaric acid Metabolic Engineering Communications Ustilago cynodontis 2-Hydroxyparaconic and itatartaric acid Derivatives specificity Metabolic engineering Fermentation Downstream processing |
| title | Selective production of the itaconic acid-derived compounds 2-hydroxyparaconic and itatartaric acid |
| title_full | Selective production of the itaconic acid-derived compounds 2-hydroxyparaconic and itatartaric acid |
| title_fullStr | Selective production of the itaconic acid-derived compounds 2-hydroxyparaconic and itatartaric acid |
| title_full_unstemmed | Selective production of the itaconic acid-derived compounds 2-hydroxyparaconic and itatartaric acid |
| title_short | Selective production of the itaconic acid-derived compounds 2-hydroxyparaconic and itatartaric acid |
| title_sort | selective production of the itaconic acid derived compounds 2 hydroxyparaconic and itatartaric acid |
| topic | Ustilago cynodontis 2-Hydroxyparaconic and itatartaric acid Derivatives specificity Metabolic engineering Fermentation Downstream processing |
| url | http://www.sciencedirect.com/science/article/pii/S221403012400021X |
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