Metabolic engineering of Deinococcus radiodurans for pinene production from glycerol
Abstract Background The objective of this work was to engineer Deinococcus radiodurans R1 as a microbial cell factory for the production of pinene, a monoterpene molecule prominently used for the production of fragrances, pharmaceutical products, and jet engine biofuels. Our objective was to produce...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2021-09-01
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| Series: | Microbial Cell Factories |
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| Online Access: | https://doi.org/10.1186/s12934-021-01674-4 |
| _version_ | 1818730845114990592 |
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| author | Seyed Hossein Helalat Carsten Jers Mandana Bebahani Hassan Mohabatkar Ivan Mijakovic |
| author_facet | Seyed Hossein Helalat Carsten Jers Mandana Bebahani Hassan Mohabatkar Ivan Mijakovic |
| author_sort | Seyed Hossein Helalat |
| collection | DOAJ |
| description | Abstract Background The objective of this work was to engineer Deinococcus radiodurans R1 as a microbial cell factory for the production of pinene, a monoterpene molecule prominently used for the production of fragrances, pharmaceutical products, and jet engine biofuels. Our objective was to produce pinene from glycerol, an abundant by-product of various industries. Results To enable pinene production in D. radiodurans, we expressed the pinene synthase from Abies grandis, the geranyl pyrophosphate (GPP) synthase from Escherichia coli, and overexpressed the native 1-deoxy-d-xylulose 5-phosphate synthase. Further, we disrupted the deinoxanthin pathway competing for the substrate GPP by either inactivating the gene dr0862, encoding phytoene synthase, or substituting the native GPP synthase with that of E. coli. These manipulations resulted in a D. radiodurans strain capable of producing 3.2 ± 0.2 mg/L pinene in a minimal medium supplemented with glycerol, with a yield of 0.13 ± 0.04 mg/g glycerol in shake flask cultures. Additionally, our results indicated a higher tolerance of D. radiodurans towards pinene as compared to E. coli. Conclusions In this study, we successfully engineered the extremophile bacterium D. radiodurans to produce pinene. This is the first study demonstrating the use of D. radiodurans as a cell factory for the production of terpenoid molecules. Besides, its high resistance to pinene makes D. radiodurans a suitable host for further engineering efforts to increase pinene titer as well as a candidate for the production of the other terpenoid molecules. |
| first_indexed | 2024-12-17T23:08:15Z |
| format | Article |
| id | doaj.art-c49f3d68a80040c098ffea55235c5251 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-12-17T23:08:15Z |
| publishDate | 2021-09-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-c49f3d68a80040c098ffea55235c52512022-12-21T21:29:12ZengBMCMicrobial Cell Factories1475-28592021-09-0120111410.1186/s12934-021-01674-4Metabolic engineering of Deinococcus radiodurans for pinene production from glycerolSeyed Hossein Helalat0Carsten Jers1Mandana Bebahani2Hassan Mohabatkar3Ivan Mijakovic4Biological Science and Technology, Isfahan UniversityThe Novo Nordisk Foundation Center for Biosustainability, Technical University of DenmarkBiological Science and Technology, Isfahan UniversityBiological Science and Technology, Isfahan UniversityThe Novo Nordisk Foundation Center for Biosustainability, Technical University of DenmarkAbstract Background The objective of this work was to engineer Deinococcus radiodurans R1 as a microbial cell factory for the production of pinene, a monoterpene molecule prominently used for the production of fragrances, pharmaceutical products, and jet engine biofuels. Our objective was to produce pinene from glycerol, an abundant by-product of various industries. Results To enable pinene production in D. radiodurans, we expressed the pinene synthase from Abies grandis, the geranyl pyrophosphate (GPP) synthase from Escherichia coli, and overexpressed the native 1-deoxy-d-xylulose 5-phosphate synthase. Further, we disrupted the deinoxanthin pathway competing for the substrate GPP by either inactivating the gene dr0862, encoding phytoene synthase, or substituting the native GPP synthase with that of E. coli. These manipulations resulted in a D. radiodurans strain capable of producing 3.2 ± 0.2 mg/L pinene in a minimal medium supplemented with glycerol, with a yield of 0.13 ± 0.04 mg/g glycerol in shake flask cultures. Additionally, our results indicated a higher tolerance of D. radiodurans towards pinene as compared to E. coli. Conclusions In this study, we successfully engineered the extremophile bacterium D. radiodurans to produce pinene. This is the first study demonstrating the use of D. radiodurans as a cell factory for the production of terpenoid molecules. Besides, its high resistance to pinene makes D. radiodurans a suitable host for further engineering efforts to increase pinene titer as well as a candidate for the production of the other terpenoid molecules.https://doi.org/10.1186/s12934-021-01674-4PineneBiofuelDeinococcus radiodurans R1Metabolic engineeringMonoterpeneGlycerol |
| spellingShingle | Seyed Hossein Helalat Carsten Jers Mandana Bebahani Hassan Mohabatkar Ivan Mijakovic Metabolic engineering of Deinococcus radiodurans for pinene production from glycerol Microbial Cell Factories Pinene Biofuel Deinococcus radiodurans R1 Metabolic engineering Monoterpene Glycerol |
| title | Metabolic engineering of Deinococcus radiodurans for pinene production from glycerol |
| title_full | Metabolic engineering of Deinococcus radiodurans for pinene production from glycerol |
| title_fullStr | Metabolic engineering of Deinococcus radiodurans for pinene production from glycerol |
| title_full_unstemmed | Metabolic engineering of Deinococcus radiodurans for pinene production from glycerol |
| title_short | Metabolic engineering of Deinococcus radiodurans for pinene production from glycerol |
| title_sort | metabolic engineering of deinococcus radiodurans for pinene production from glycerol |
| topic | Pinene Biofuel Deinococcus radiodurans R1 Metabolic engineering Monoterpene Glycerol |
| url | https://doi.org/10.1186/s12934-021-01674-4 |
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