Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Impact of Carbon Fixation, Distribution and Storage on Terpene Production in Euryhaline Cyanobacteria

Version 1 : Received: 17 November 2023 / Approved: 20 November 2023 / Online: 20 November 2023 (16:06:55 CET)

How to cite: Vincent, M.; Blanc-Garin, V.; Chenebault, C.; Cirimele, M.; Farci, S.; Garcia-Alles, L.F.; Cassier-Chauvat, C.; Chauvat, F. Impact of Carbon Fixation, Distribution and Storage on Terpene Production in Euryhaline Cyanobacteria. Preprints 2023, 2023111252. https://doi.org/10.20944/preprints202311.1252.v1 Vincent, M.; Blanc-Garin, V.; Chenebault, C.; Cirimele, M.; Farci, S.; Garcia-Alles, L.F.; Cassier-Chauvat, C.; Chauvat, F. Impact of Carbon Fixation, Distribution and Storage on Terpene Production in Euryhaline Cyanobacteria. Preprints 2023, 2023111252. https://doi.org/10.20944/preprints202311.1252.v1

Abstract

Terpenes are high-value chemicals which can be produced by engineered cyanobacteria from sustainable resources, solar energy, water and CO2. We previously reported that the euryhaline unicellular cyanobacteria Synechocystis PCC 6803 (S.6803) and Synechococcus PCC 7002 (S.7002) produce farnesene and limonene, respectively, more efficiently than other terpenes. In the present study, we attempted at enhancing farnesene production in S.6803 and limonene production in S.7002. Practically, we tested the influence of key cyanobacterial enzymes acting in carbon fixation (RubisCO, PRK, CcmK3 and CcmK4), utilization (CrtE, CrtR and CruF) and storage (PhaA and PhaB) on terpene production in S.6803, and we compared some of the findings with the data obtained in S.7002. We report that the overproduction of RubisCO from S.7002 and PRK from Cyanothece PCC 7425 increased farnesene production in S.6803, but not limonene production in S.7002. The overexpression of the crtE genes (synthesis or terpene precursors) from S.6803 or S.7002 did not increased farnesene production in S.6803. In contrast, the overexpression of the crtE gene from S.6803, but not S.7002, increased farnesene production in S.7002, emphasizing the physiological difference between these two model cyanobacteria. Furthermore, the deletion of the crtR and cruF genes (carotenoid synthesis) and phaAB genes (carbon storage) did not increase the production of farnesene in S.6803. Finally, as a containment strategy of genetically modified (GMO) strains of S.6803 we report that the deletion of the ccmK3K4 genes (carboxysome for CO2 fixation) did not affect the production of limonene but decreased the production of farnesene in S.6803.

Keywords

Synechocystis PCC 6803; Synechococcus PCC 7002; farnesene; limonene; RubisCO; phosphoribulokinase; carboxysome; geranylgeranyl pyrophosphate synthase; β-carotene hydroxylase; γ-carotene hydroxylase

Subject

Biology and Life Sciences, Biology and Biotechnology

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