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Lutein, made by the α-branch of the methyl-erythritol phosphate (MEP) pathway, is one of the most abundant xanthophylls in plants. It is involved in the structural stabilization of light-harvesting complexes, transfer of excitation energy to chlorophylls and photoprotection. In contrast, lutein and the α-branch of the MEP pathway are not present in cyanobacteria. In this study, we genetically engineered the cyanobacterium  Synechocystis  for the missing MEP α-branch resulting in lutein accumulation. A cassette comprising four  Arabidopsis thaliana  genes coding for two lycopene cyclases ( AtLCYe  and  AtLCYb ) and two hydroxylases ( AtCYP97A  and  AtCYP97C ) was introduced into a  Synechocystis  strain that lacks the endogenous, cyanobacterial lycopene cyclase  cruA . The resulting  synlut  strain showed wild-type growth and only moderate changes in total pigment composition under mixotrophic conditions, indicating that the  cruA  deficiency can be complemented by  Arabidopsis  lycopene cyclases leaving the endogenous β-branch intact. A combination of liquid chromatography, UV-Vis detection and mass spectrometry confirmed a low but distinct synthesis of lutein at rates of 4.8 ± 1.5 nmol per liter culture at OD 730  (1.03 ± 0.47 mmol mol –1  chlorophyll). In conclusion,  synlut  provides a suitable platform to study the α-branch of the plastidic MEP pathway and other functions related to lutein in a cyanobacterial host system.

Introduction of the Carotenoid Biosynthesis α-Branch Into Synechocystis sp. PCC 6803 for Lutein Production