Triplet energy transfer in bacterial photosynthetic reaction centres

[3-vinyl]-13(2)-OH-bacteriochlorophyll a has been selectively exchanged against native bacteriochlorophyll a in the monomer binding sites at the A- and B-branch of the photosynthetic reaction centre from Rhodobacter sphaeroides. Transient absorption difference measurements were performed on these samples over a temperature range from 4.2 to 300 K with 20 ns time resolution. Specifically the decay of the primary donor triplet state, 3P870, as well as the rise and decay rates of the carotenoid triplet state. 3Car (spheroidene), were measured. The observed rates revealed a thermally activated carotenoid triplet formation corresponding to the decay of the primary donor triplet state. The activation energies for the triplet energy transfer process were 100 (+/-10) cm-1 for reaction centers from wild-type Rhodobacter sphaeroides 2.4.1, with and without exchange of the monomeric bacteriochlorophyll on the electron transfer-active branch, BA. For reaction centers from Rhodobacter sphaeroides R26.1 with both monomers exchanged against [3-vinyl]-13(2)-OH-bacteriochlorophyll a, and subsequent spheroidene reconstitution the activation energy was 460 (+/-20) cm-1. These activation energies correspond to the energy difference between the triplet states of the accessory BChl monomer, BB, and the primary donor when native BChl a or [3-vinyl]-13(2)-OH-BChl a is present in the BB binding site. In all samples the 3Car formation rates were bi-phasic over a large temperature range. A fast temperature-independent rate was observed on the wavelength of the carotenoid triplet-triplet absorption which dominated at very low temperatures. Additionally, a slower temperature-independent 3Car formation rate was observed at low temperatures which could be explained with the assumption of heterogeneity in the energy barrier (3BB) and/or the primary donor triplet state (3P870)) A tunneling mechanism as proposed earlier by Kolaczkowski (PhD thesis, Brown University. 1989) is not only unnecessary but also incompatible with the available experimental data.