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Chen, Que; Arents, Jos; Schuurmans, J. Merijn; Ganapathy, Srividya; de Grip, Willem J.; Cheregi, Otilia; Funk, Christiane; dos Santos, Filipe Branco; Hellingwerf, Klaas J.
Metabolic engineering, March 2019, 2019-03-00, 20190301, Letnik: 52Journal Article
To fill the “green absorption gap”, a green absorbing proteorhodopsin was expressed in a PSI-deletion strain (ΔPSI) of Synechocystis sp. PCC6803. Growth-rate measurements, competition experiments and physiological characterization of the proteorhodopsin-expressing strains, relative to the ΔPSI control strain, allow us to conclude that proteorhodopsin can enhance the rate of photoheterotrophic growth of ΔPSI Synechocystis strain. The physiological characterization included measurement of the amount of residual glucose in the spent medium and analysis of oxygen uptake- and production rates. To explore the use of solar radiation beyond the PAR region, a red-shifted variant Proteorhodopsin-D212N/F234S was expressed in a retinal-deficient PSI-deletion strain (ΔPSI/ΔSynACO). Via exogenous addition of retinal analogue an infrared absorbing pigment (maximally at 740 nm) was reconstituted in vivo. However, upon illumination with 746 nm light, it did not significantly stimulate the growth (rate) of this mutant. The inability of the proteorhodopsin-expressing ΔPSI strain to grow photoautotrophically is most likely due to a kinetic rather than a thermodynamic limitation of its NADPH-dehydrogenase in NADP+-reduction. •Proteorhodopsin expression increases growth rate of a ∆PSI Synechocystis strain with at least 16 %.•Proteorhodopsin contributes to proton motive force generation in vivo in Synechocystis.•A transgenic Synechocystis strain has been generated that can absorb infrared light.•‘Reversed electron transfer’ through NDH-1 could not be demonstrated in Synechocystis.
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