Journal article
Metabolic engineering of Deinococcus radiodurans for pinene production from glycerol
Department of Health Technology, Technical University of Denmark1
Nanomaterials and Nanobiosensors, Biotherapeutic Engineering and Drug Targeting, Department of Health Technology, Technical University of Denmark2
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark3
Bacterial Signal Transduction, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark4
University of Isfahan5
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.
Language: | English |
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Publisher: | BioMed Central |
Year: | 2021 |
Pages: | 187 |
ISSN: | 14752859 |
Types: | Journal article |
DOI: | 10.1186/s12934-021-01674-4 |
ORCIDs: | 0000-0003-3240-2672 , Helalat, Seyed Hossein , Jers, Carsten and Mijakovic, Ivan |