A cytochrome c551 mediates the cyclic electron transport chain of the anoxygenic phototrophic bacterium Roseiflexus castenholzii

Roseiflexus castenholzii is a gram-negativefilamentous phototrophic bacterium that carries out anoxygenic photosynthesis through a cyclic electron transport chain(ETC).The ETC is composed of a reaction center(RC)–light-harvesting(LH)complex(rcRC–LH);an alternative complex III(rcACIII),which functionally re-places the cytochrome bc1/b6f complex;and the periplasmic electron acceptor auracyanin(rcAc).Although compositionally and structurally different from the bc1/b6f complex,rcACIII plays similar essential roles in oxidizing menaquinol and transferring electrons to the rcAc.However,rcACIII-mediated electron transfer(which includes both an intraprotein route and a downstream route)has not been clearly elucidated,nor have the details of cyclic ETC.Here,we identify a previously unknown monoheme cytochrome c(cyt c551)as a novel periplasmic electron acceptor of rcACIII.It reduces the light-excited rcRC–LH to complete a cyclic ETC.We also reveal the molecular mechanisms involved in the ETC using electron paramagnetic resonance(EPR),spectroelectrochemistry,and enzymatic and structural analyses.Wefind that electrons released from rcACIII-oxidized menaquinol are transferred to two alternative periplasmic electron acceptors(rcAc and cyt c551),which eventually reduce the rcRC to form the complete cyclic ETC.This work serves as a foundation for further studies of ACIII-mediated electron transfer in anoxygenic photosynthesis and broadens our under-standing of the diversity and molecular evolution of prokaryotic ETCs.