Metabolic Engineering of Deinococcus radiodurans for the Production of Phytoene

Abstract

A metabolically-engineered Deinococcus radiodurans R1 strain capable of producing phytoene, a colorless C 4 0 carotenoid and a promising antioxidant, has been developed. To make this base strain, first, the crtI gene encoding phytoene desaturase was deleted to block the conversion of phytoene to other carotenoids such as lycopene and γ-carotene. This engineered strain produced 0.413 ± 0.023 mg/l of phytoene from 10 g/l of fructose. Further enhanced production of phytoene up to 4.46 ± 0.19 mg/l was achieved by overexpressing the crtB gene encoding phytoene synthase and the dxs genes encoding 1-deoxy-D-xylulose-5-phosphate synthase gene, and by deleting the crtD gene. High cell-density culture of our final engineered strain allowed production of 10.3 ± 0.85 mg/l of phytoene with the yield and productivity of 1.04 ± 0.05 mg/g and 0.143 ± 0.012 mg/l/h, respectively, from 10 g/l of fructose. Furthermore, the antioxidant potential of phytoene produced by the final engineered strain was confirmed by in vitro DPPH radical-scavenging assay.

Isolation of Lactobacillus plantarum subsp. plantarum Producing C30 Carotenoid 4,4��-Diaponeurosporene and the Assessment of Its Antioxidant Activity

“…On the other hand, antioxidants such as carotenoids are substances that can reduce ROS and prevent macromolecule oxidation[30]. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging method has been used to evaluate the antioxidant capacity of carotenoids extracted from microorganisms[31]. C 5 0…”

Active Transposition of Insertion Sequences by Oxidative Stress in Deinococcus geothermalis

“…In accordance with these needs, several prokaryotic strains were applied to the production of carotenoid intermediates by target gene disruption. It has been reported that some microorganisms like Thermococcus kodakarensis can produce phytoene of approximately 2.6 mg/L, and 10.4 mg/L for D. radiodurans (Fuke et al, 2018;Jeong et al, 2018). Thus, this gene disruption approach of carotenoid gene by IS transposition event might be used for the bio-production of useful substances in the near future.…”

Metabolic engineering of Deinococcus radiodurans for pinene production from glycerol

“…Further, harboring an optimized mevalonate pathway, enhancement of upstream pathways, and the potential of optimizing growth conditions during production should be evaluated. It has been shown that the use of high cell-density cultures for carotenoid production in D. radiodurans could increase the production of terpene molecules significantly [ 21 , 32 ], and this approach can be used for pinene production as well.…”

“…Deinococcus radiodurans is known to be extremely tolerant to different sources of stress: ionizing radiation, reactive oxygen species, UV light, heavy metals, dryness, high salinity, and alcohols. Additionally, it is able to utilize various carbon sources (different sugars, glycerol, fats, and proteins) [ 18 , 19 ], making it a promising candidate for various purposes such as bioremediation, decomposition of toxic compounds, and production of valuable molecules [ 20 , 21 ]. Considering that manganese is the main cofactor of most pinene synthases, including that of A. grandis , the presence of high levels of manganese in D. radiodurans (0.2–4 mM) is another positive feature [ 22 ].…”