期刊
MICROBIAL CELL FACTORIES
卷 20, 期 1, 页码 -出版社
BMC
DOI: 10.1186/s12934-021-01674-4
关键词
Pinene; Biofuel; Deinococcus radiodurans R1; Metabolic engineering; Monoterpene; Glycerol
资金
- Novo Nordisk Foundation [NNF10CC1016517]
This study successfully engineered Deinococcus radiodurans R1 to produce pinene using glycerol as a substrate. Through gene expression and metabolic pathway disruption, a strain capable of producing pinene was developed. Additionally, it was found that Deinococcus radiodurans exhibited higher tolerance towards pinene compared to Escherichia coli.
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.
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