Abstract
BackgroundThe 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.ResultsTo 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.ConclusionsIn 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.
Highlights
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
Potential of D. radiodurans as a pinene production host In the present study, we investigated the potential of using D. radiodurans as a novel host for the production of the monoterpene molecule pinene using glycerol as a substrate
We hypothesized that D. radiodurans could be well suited due to the fact that it produces a pigment, deinoxanthin, that like pinene is synthesized from geranyl pyrophosphate (GPP)
Summary
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. The human population will exceed 9 billion by 2050, which leads to Glycerol is a by-product of various large-scale industrial processes, such as production of soap (saponification of triglycerides) and biodiesel (transesterification). It has been proposed as a cheap carbon source for fermentation [6]. Glycerol is a more reduced carbon source, delivering more reducing equivalents per unit price [7] This polyol compound is a convenient carbon source for producing valuable molecules using microbial cell factories
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