title: Accelerated electron transport from photosystem I to redox partners by covalently linked ferredoxin.
abstract: Photosystem I is a highly efficient and potent light-induced reductase that is considered to be an appealing target for integration into hybrid solar fuel production systems. However, rapid transport of multiple electrons from the reducing end of photosystem I to downstream processes in vivo is limited by the diffusion of its native redox partner ferredoxin that is a single electron carrier. Here, we describe the design and construction of a faster electron transfer interface based on anchoring ferredoxin to the reducing end of photosystem I thereby confining the diffusion space of ferredoxin to the near vicinity of its photosystem I binding and reduction site. This was achieved by fusing ferredoxin to the PsaE subunit of photosystem I by a flexible peptide linker and reconstituting PSI in vitro with the new fusion protein. A computational algorithm was developed in order to determine the optimal linker length that will confine ferredoxin to the vicinity of photosystem I's reducing end without restricting the formation of electron transfer complexes. According to the calculation, we reconstituted photosystem I with three fusion proteins comprising PsaE and ferredoxin separated by linkers of different lengths, namely 14, 19, and 25 amino acids, and tested their effect on electron transfer rates from photosystem I to downstream processes. Indeed, we found a significant enhancement of light dependent NADPH synthesis using photosystems containing the PsaE-ferredoxin fusion proteins, equivalent to a ten-fold increase in soluble ferredoxin concentration. We propose that such a system could be used for other ferredoxin dependent redox reactions, such as the enzymatic production of hydrogen, a promising alternative fuel. As the system is comprised entirely of natural amino acids and biological cofactors, it could be integrated into the energy conversion apparatus of photosynthetic organisms by genetic engineering.
authors.affiliation: Plant Sciences Department, Weizmann Institute of Science, Rehovot 7610001, Israel.
authors.forename: Gal
authors.initials: G
authors.lastname: Wittenberg
authors.forename: William
authors.initials: W
authors.lastname: Sheffler
authors.forename: Dana
authors.initials: D
authors.lastname: Darchi
authors.forename: David
authors.initials: D
authors.lastname: Baker
authors.forename: Dror
authors.initials: D
authors.lastname: Noy
mesh_terms: D004579:Electron Transport; D005288:Ferredoxins / Q000378:metabolism*; D007700:Kinetics; D008958:Models, Molecular; D009249:NADP / Q000378:metabolism; D045331:Photosystem I Protein Complex / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D011487:Protein Conformation; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D046940:Synechococcus / Q000235:genetics
chemical_list: D005288:Ferredoxins; D045331:Photosystem I Protein Complex; D011993:Recombinant Fusion Proteins; C079741:photosystem I, psaE subunit; D009249:NADP