Metabolic Engineering of Deinococcus radiodurans for the Production of Phytoene.

Abstract

A metabolically-engineered Deinococcus radiodurans R1 strain capable of producing phytoene, a colorless C40 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.