Construction and characterization of genetically modified synechocystis sp. PCC 6803 photosystem II core complexes containing carotenoids with shorter pi-conjugation than beta-carotene.
Beta-carotene has been identified as an intermediate in a secondary electron transfer pathway that oxidizes Chl(Z) and cytochrome b(559) in Photosystem II (PS II) when normal tyrosine oxidation is blocked. To test the redox function of carotenoids in this pathway, we replaced the zeta-carotene desaturase gene (zds) or both the zds and phytoene desaturase (pds) genes of Synechocystis sp. PCC 6803 with the phytoene desaturase gene (crtI) of Rhodobacter capsulatus, producing carotenoids with shorter conjugated pi-electron systems and higher reduction potentials than beta-carotene. The PS II core complexes of both mutant strains contain approximately the same number of chlorophylls and carotenoids as the wild type but have replaced beta-carotene (11 double bonds), with neurosporene (9 conjugated double bonds) and beta-zeacarotene (9 conjugated double bonds and 1 beta-ionylidene ring). The presence of the ring appears necessary for PS II assembly. Visible and near-infrared spectroscopy were used to examine the light-induced formation of chlorophyll and carotenoid radical cations in the mutant PS II core complexes at temperatures from 20 to 160 K. At 20 K, a carotenoid cation radical is formed having an absorption maximum at 898 nm, an 85 nm blue shift relative to the beta-carotene radical cation peak in the WT, and consistent with the formation of the cation radical of a carotenoid with 9 conjugated double bonds. The ratio of Chl(+)/Car(+) is higher in the mutant core complexes, consistent with the higher reduction potential for Car(+). As the temperature increases, other carotenoids become accessible to oxidation by P(680)(+).
Bautista
James A
JA
CR&D, Experimental Station, E. I. du Pont de Nemours & Co., Wilmington, Delaware 19880-0173, USA.
Tracewell
Cara A
CA
Schlodder
Eberhard
E
Cunningham
Francis X
FX
Brudvig
Gary W
GW
Diner
Bruce A
BA
D002338:Carotenoids / Q000737:chemistry*; D002412:Cations; D002734:Chlorophyll / Q000737:chemistry; D002845:Chromatography; D002851:Chromatography, High Pressure Liquid; D004583:Electrons; D017353:Gene Deletion; D008027:Light; D008345:Manganese / Q000737:chemistry; D008956:Models, Chemical; D008958:Models, Molecular; D009154:Mutation; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000378:metabolism; D010100:Oxygen / Q000737:chemistry / Q000378:metabolism; D045322:Photosynthetic Reaction Center Complex Proteins / Q000502:physiology; D045332:Photosystem II Protein Complex / Q000737:chemistry*; D010858:Pigmentation; D016354:Rhodobacter capsulatus / Q000378:metabolism; D013053:Spectrophotometry; D013055:Spectrophotometry, Infrared; D046939:Synechocystis / Q000235:genetics* / Q000378:metabolism*; D013696:Temperature; D013997:Time Factors; D014443:Tyrosine / Q000737:chemistry; D019207:beta Carotene / Q000737:chemistry / Q000378:metabolism*
D002412:Cations; D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; D019207:beta Carotene; D002734:Chlorophyll; D002338:Carotenoids; D014443:Tyrosine; D008345:Manganese; D010088:Oxidoreductases; C022398:phytoene dehydrogenase; C099215:zeta-carotene desaturase; D010100:Oxygen