Metabolic engineering of Deinococcus radiodurans for pinene production from glycerol

Seyed Hossein Helalat, Carsten Jers, Mandana Bebahani*, Hassan Mohabatkar, Ivan Mijakovic

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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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.

Original languageEnglish
Article number187
JournalMicrobial Cell Factories
Volume20
Issue number1
Number of pages14
ISSN1475-2859
DOIs
Publication statusPublished - Dec 2021

Bibliographical note

Funding Information:
This work was supported by a Grant from the Novo Nordisk Foundation (NNF10CC1016517) to IM.

Funding Information:
We thank Aaron John Christian Andersen, metabolomics core manager at the department of biotechnology and biomedicine (Technical University of Denmark), for his assistance in GC-MS analysis.

Publisher Copyright:
© 2021, The Author(s).

Keywords

  • Biofuel
  • Deinococcus radiodurans R1
  • Glycerol
  • Metabolic engineering
  • Monoterpene
  • Pinene

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