Metabolic engineering of Deinococcus radiodurans for pinene

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.

Frontiers Deinococcus radiodurans UWO298 Dependence on Background Radiation for Optimal Growth

The Toughest Organisms on Earth: Evolution of Radio-Resistance Is More Complicated Than Previously Thought

Biosynthesis, evolution and ecology of microbial terpenoids - Natural Product Reports (RSC Publishing) DOI:10.1039/D1NP00047K

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Functionalized Nanomaterial Assembling and Biosynthesis Using the Extremophile Deinococcus radiodurans for Multifunctional Applications - Li - 2019 - Small - Wiley Online Library

Microbial Synthesis of Pinene

Engineering Robustness of Microbial Cell Factories - Gong - 2017 - Biotechnology Journal - Wiley Online Library

Microbial Cell Factories

Metabolic engineering of Deinococcus radiodurans for pinene production from glycerol, Microbial Cell Factories

Metabolic engineering of Deinococcus radiodurans for pinene production from glycerol, Microbial Cell Factories

PDF] Metabolic Engineering of Deinococcus radiodurans for the Production of Phytoene.

Frontiers Lack of the Bacterial Phytochrome Protein Decreases Deinococcus radiodurans Resistance to Mitomycin C

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