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Summary Photo-generation of a proton gradient over a lipid bilayer is of interest due to its essential role in photosynthetic bacteria. Membrane asymmetry is key to the proton gradient generation via directional proton transport. Here, we report a light-driven proton pump based on two-dimensional, porphyrin-based Janus metal-organic layers (Janus-MOLs). The Janus-MOL, functionalized with carboxyquinone on one side and Acitretin on the other via a microemulsion-based method, was attached to liposome surface. Upon photoexcitation, the porphyrins initiate electron and hole transfers to carboxyquinone and Acitretin, respectively, which undergo redox reactions with freely diffusing quinone (Q)/hydrosemiquinone (HQ·) in the lipid bilayer to produce a concentration gradient of quinone-based species. Owing to different pKa values of HQ+ and HQ·, these redox reactions trigger proton transport across the membrane to create a pH gradient, which drives ATP production by CFoF1-ATP synthase in a similar fashion as photosynthetic bacteria.

Control of the spectral overlap between energy donors and acceptors provides insight into excitation energy transfer (EET) mechanisms in photosynthetic light-harvesting proteins. Substitution of energy-donating B800 bacteriochlorophyll (BChl) a with other pigments in the light-harvesting complex 2 (LH2) of purple photosynthetic bacteria has been extensively performed; however, most studies on the B800 substitution have focused on the decrease in the spectral overlap integral with energy-accepting B850 BChl a by reconstitution of chlorophylls into the B800 site. Here, we reconstitute BChl b into the B800 site of the LH2 protein from Rhodoblastus acidophilus to increase the spectral overlap with B850 BChl a. BChl b in the B800 site had essentially the same hydrogen-bonding pattern as B800 BChl a, whereas it showed a red-shifted Qy absorption band at 831 nm. The EET rate from BChl b to B850 BChl a in the reconstituted LH2 was similar to that of native LH2 despite the red shift of the Qy band of the energy donor. These results demonstrate the importance of the contribution of the density of excitation states of the B850 circular assembly, which incorporates higher lying optically forbidden states, to intracomplex EET in LH2.

Utilizing solar energy for hydrogen production by combining light-activated materials and biocatalysts has become a promising alternative to fossil fuels. Herein, a feasible and simple bio-hybrid complex based on water-soluble oligofluorene (OF), polythiophene (PTP) and Rhodopseudomonas palustris (R. palustris), one kind of photosynthetic bacteria, was constructed for enhancing photocatalytic hydrogen production. Under the irradiation of visible light, there was fluorescence resonance energy transfer (FRET) between OF and PTP, which amplified photoelectron signals and transferred electrons via PTP to methyl viologen (MV2+), an electron mediator that established electron transfer pathways between organic materials and R. palustris. Hydrogen production involving R. palustris was catalyzed by nitrogenase. It is noteworthy that MV2+ was reduced to MV+˙ and transferred electrons to nitrogenase in R. palustris, converting protons and electrons to molecular hydrogen. Besides, triethanolamine (TEOA) was added as an electron donor to provide electrons to PTP. The proposed OF/PTP/MV2+/TEOA/R. palustris system provided a feasible strategy for photocatalytic hydrogen production with low-cost and easily implemented techniques.

Scanning spectrophotometers equipped with integrating spheres were once highly specialized spectrophotometric equipment but are now more readily available commercially. They are particularly useful for objectively assessing the spectral absorption and reflectance of algal cell suspensions. Ordinary dual-beam spectrometers do not give valid measurements of the spectral properties of cells due to light scattering. Spectra of unicellular algae using integrating sphere spectroscopy can measure the absorbance (Abs), transmissions (%T) and reflectance (%R) and hence the actual absorptance (%Abt) of turbid cell suspensions and hence the in vivo pigment absorption properties of photosynthetic organisms. These results were compared with those obtained using conventional dual-beam spectrophotometry scans on turbid cell suspensions and the in solvent spectra of photosynthetic pigments. The common unicellular green alga, Chlorella sp., is used as an example of an oxygenic photo-organism with chlorophyll a as the primary photosynthetic pigment and comparisons made to other unicellular algae such as a cyanobacterium (Synechococcus), Acaryochloris and a diatom (Chaetoceros). Photosynthetic bacteria, such as Rhodopseudomonas palustris, are photosynthetic but do not produce oxygen, and their photosynthesis is usually based on bacteriochlorophyll a. Comparisons are made of integrating sphere vs. dual-beam transmission spectroscopy of BChl a and BChl b organisms in solvent and in vivo of anoxygenic photosynthetic bacteria (Afifella & Thermochromatium [BChl a], Blastochloris [BChl b]) and with oxygenic organisms.

Oxygenic photosynthesis can be measured easily using O2 or CO2 gas exchange, oxygen electrodes, Winkler titration, 14CO2-fixation and by PAM (pulse amplitude modulation) fluorometry. PAM estimates the photosynthetic electron transport rate (ETR) by measuring the variable fluorescence of chlorophyll (Chl) a (> 695 nm) induced by absorption of blue or red light. Anoxygenic photosynthetic bacteria (APB) do not use water as an electron source and are typically photoheterotrophic rather than photoautotrophic and so 14CO2 fixation is a misleading estimate of photosynthetic electron transport in APB photosynthesis. In vivo bacteriochlorophyll a (BChl a) absorbs blue light similar to Chl a but its characteristic longer-wavelength absorption is in the infrared and fluorescence is at > 800 nm. Blue light-induced PAM fluorescence can be used to measure the ETR in purple non-sulphur anoxygenic photobacteria and purple sulphur photobacteria because their RC-2 type BChl a complexes fluoresce similarly to PSII but at longer wavelengths than Chl a. Conventional PAM fluorometers using blue light cannot readily distinguish between oxygenic and RC-2 type anoxygenic photosynthesis because they use a simple > 700 nm highpass filter in front of the detector diode. We modified one fluorometer to use a 695–750-nm bandpass filter to measure Chl a fluorescence from PS-II, representing oxygenic photosynthesis. Similarly, we modified another fluorometer to use a highpass filter (> 830 nm) to measure BChl a fluorescence, representing anoxygenic photosynthesis. However, the fluorescence bands of Chl a and BChl a were found to be too wide to unambiguously distinguish between oxygenic and anoxygenic photosynthesis purely by fluorometry. Treatment with the specific PS-II inhibitor DCMU (Diuron) did enable discrimination of the two types of photosynthesis in a mixture of oxygenic and anoxygenic organisms. Ecological niches made up of both oxygenic and anoxygenic organisms such as microbial mats and hypereutrophic environments such as sewage ponds, wastewater ponds and prawn farm ponds are much more common than often realized. Anoxygenic photosynthesis in such systems is significant yet largely unquantified.

The possibility of singlet oxygen generation in five species of photosynthetic bacteria under irradiation with light was studied using the Singlet Oxygen Sensor Green (SOSG) dye that, when singlet oxygen is bound, is transformed into a fluorescent endoperoxide form. Irradiation was carried out using light of different spectral composition: red (absorption of bacteriochlorophyll, BChl) and white (absorption of BChl and carotenoids). No singlet oxygen generation was detected under red light irradiation, which indicated that BChl was not involved in this process. Under white light, as the irradiation time increased, an increase in SOSG fluorescence was recorded in the membranes of four bacterial species: Allochromatium vinosum MSU, Rhodobacter sphaeroides G1C, Rba. blasticus K-1 and Rhodopseudomonas faecalis. In Rba. sphaeroides, no increase in SOSG fluorescence was observed. It is assumed that in the white light this process occurs due to the pigments that absorb in the blue-green region of the spectrum, i.e. carotenoids with 8‒11 conjugated double bonds (neurosporene, spheroidene, lycopene, and rhodopin). Similar to Trolox, SOSG may slow down the bleaching process of BChl850 in the membranes of Alc. vinosum strain MSU and may therefore effectively perform the role of a singlet oxygen trap.

Living supramolecular polymerization is a breakthrough system to control the size of supramolecular aggregates. Multidimensional (2D or 3D) self-aggregates of chiral and unsymmetric molecules are observed in nature. For instance, a chlorosome, which is the main light-harvesting antenna in green photosynthetic bacteria, possesses tubular structures of chlorophyll pigments with J-type slipped cofacial and circular arrangements. Here, we report size-controllable construction of chlorosome-like aggregates by photoactivated supramolecular assembly of “caged chlorophyll”. The caged chlorophylls, which were nonassembling (inert) species, were illuminated with UV-light to give the active species one by one, and then the resulting active monomers spontaneously assembled to construct tubular self-aggregates. The length of such chlorosome-like aggregate tubes was dependent on the UV-irradiation duration and intensity of UV-light. The photoactivated supramolecular assembly system would be useful for application of (o...

The effect of singlet oxygen on light-harvesting (LH) complexes has been studied for a number of sulfur (S+) and nonsulfur (S−) photosynthetic bacteria. The visible/near-IR absorption spectra of the standard LH2 complexes (B800-850) of Allochromatium (Alc.) vinosum (S+), Rhodobacter (Rba.) sphaeroides (S−), Rhodoblastus (Rbl.) acidophilus (S−), and Rhodopseudomonas (Rps.) palustris (S−), two types LH2/LH3 (B800-850 and B800-830) of Thiorhodospira (T.) sibirica (S+), and an unusual LH2 complex (B800-827) of Marichromatium (Mch.) purpuratum (S+) or the LH1 complex from Rhodospirillum (Rsp.) rubrum (S−) were measured in aqueous buffer suspensions in the presence of singlet oxygen generated by the illumination of the dye Rose Bengal (RB). The content of carotenoids in the samples was determined using HPLC analysis. The LH2 complex of Alc. vinosum and T. sibirica with a reduced content of carotenoids was obtained from cells grown in the presence of diphenylamine (DPA), and LH complexes were obtained from the carotenoidless mutant of Rba. sphaeroides R26.1 and Rps. rubrum G9. We found that LH2 complexes containing a complete set of carotenoids were quite resistant to the destructive action of singlet oxygen in the case of Rba. sphaeroides and Mch. purpuratum. Complexes of other bacteria were much less stable, which can be judged by a strong irreversible decrease in the bacteriochlorophyll (BChl) absorption bands (at 850 or 830 nm, respectively) for sulfur bacteria and absorption bands (at 850 and 800 nm) for nonsulfur bacteria. Simultaneously, we observe the appearance of the oxidized product 3-acetyl-chlorophyll (AcChl) absorbing near 700 nm. Moreover, a decrease in the amount of carotenoids enhanced the spectral stability to the action of singlet oxygen of the LH2 and LH3 complexes from sulfur bacteria and kept it at the same level as in the control samples for carotenoidless mutants of nonsulfur bacteria. These results are discussed in terms of the current hypothesis on the protective functions of carotenoids in bacterial photosynthesis. We suggest that the ability of carotenoids to quench singlet oxygen (well-established in vitro) is not well realized in photosynthetic bacteria. We compared the oxidation of BChl850 in LH2 complexes of sulfur bacteria under the action of singlet oxygen (in the presence of 50 μM RB) or blue light absorbed by carotenoids. These processes are very similar: {[BChl + (RB or carotenoid) + light] + O2} → AcChl. We speculate that carotenoids are capable of generating singlet oxygen when illuminated. The mechanism of this process is not yet clear.

The interactions of chlorophyll (Chl) and bacteriochlorophyll (BChl) pigments with the polypeptides in photosynthetic light-harvesting proteins are responsible for controlling the absorption energy of (B)Chls in protein matrixes. The binding pocket of B800 BChl a in LH2 proteins, which are peripheral light-harvesting proteins in purple photosynthetic bacteria, is useful for studying such structure-property relationships. We report the reconstitution of Chl f, which has the formyl group at the 2-position, in the B800 cavity of LH2 from the purple bacterium Rhodoblastus acidophilus. The Qy absorption band of Chl f in the B800 cavity was shifted by 14 nm to longer wavelength compared to that of the corresponding five-coordinated monomer in acetone. This redshift was larger than that of Chl a and Chl b. Resonance Raman spectroscopy indicated hydrogen bonding between the 2-formyl group of Chl f and the LH2 polypeptide. These results suggest that this hydrogen bonding contributes to the Qy redshift of Chl f. Furthermore, the Qy redshift of Chl f in the B800 cavity was smaller than that of Chl d. This may have arisen from the different patterns of hydrogen bonding between Chl f and Chl d and/or from the steric hindrance of the 3-vinyl group in Chl f.

The direct action of singlet oxygen on the bacteriochlorophyll (BChl) of light-harvesting complexes in the membranes of four types of purple non-sulfur and sulfur photosynthesizing bacteria with and without carotenoids has been studied. It has been found that BChl in carotenoid-less samples is generally more resistant to the action of singlet oxygen compared to the control. It is assumed that carotenoids are not needed to protect BChl of bacterial light-harvesting complexes from singlet oxygen, and in the classic work of Griffith et al. [1] the apoptosis process in carotenoid-less mutant cells, which involves the destruction of complexes, the appearance of monomeric BChl and generation of singlet oxygencaused by BChl, followed by BChl oxidation, was mistakenly attributed to the protective function of carotenoids.

Photo-generation of a proton gradient over a lipid bilayer is of interest due to its essential role in photosynthetic bacteria. Membrane asymmetry is key to the generation of a proton gradient via directional proton transport. Here we report a light-driven proton pump based on two-dimensional, porphyrin-based Janus metal-organic layers (Janus-MOLs) embedded in liposomes. We developed a microemulsion-based method to functionalize the Janus-MOLs with carboxyquinone on one side and Acitretin on the other side. By attaching the Janus-MOLs to liposome surfaces, we obtained a mimic to photosynthetic bacteria. Upon photoexcitation, the porphyrins initiate electron and hole transfers to carboxyquinone and Acitretin, respectively, which undergo redox reactions with freely diffusing quinone (Q) / hydrosemiquinone (HQ·) in the lipid bilayer to produce a concentration gradient of quinone-based species. Owing to different pKa values of HQ+ and HQ ·, these redox reactions trigger proton transport across the membrane to create a pH gradient, which drives ATP production by CF0F1-ATP synthase in a similar fashion as photosynthetic bacteria.

Photoactive bio-hybrid soft nanoparticles are obtained by embedding the Rhodobacter sphaeroides Reaction Center (RC) in polydopamine (PDA) suspended aggregates and treating them with ethylenediamine (EDA). Such PDA:EDA@RC nanoparticles are investigated for photocurrent generation in photoelectrochemical cells, capable to convert sunlight into electrical energy. The photosynthetic protein retains its structural and functional integrity in the nanostructures and the PDA:EDA@RC nanoparticles exhibit better water dispersity, improved light collection ability and higher photocurrent generation compared to the PDA@RC precursors, where the reaction center is embedded in pure PDA. The hybrid soft nanoparticles incorporating the RC bacterial photoenzyme show charge separated state generation comparable to that of the pristine enzyme in solution, overcoming the main limitation of RC encapsulation in pure PDA, which is the polymer low light transmission ability. As a consequence, photocurrents obtained by RC within the PDA:EDA environment are almost doubled with respect to the PDA@RC. The bio-hybrid composites described here represent an interesting example of effective functional nanostructures for sunlight photoconversion based on a biological component addressed in a tunable bio-compatible polymer composite, also showing the potentialities of fine chemical tailoring of polydopamine bio-interfaces.

Photosynthetic bacteria contain different photosynthetic pigments and can convert sunlight into the energy they need for their metabolism and to release hydrogen. However, they cannot absorb light in all bands of sunlight and can absorb only specific bands. Filtering technology is used to couple solar energy entering the system with the absorption spectral characteristics of photosynthetic hydrogen-producing bacteria, effectively avoiding the “light saturation effect”. The cell morphology of photosynthetic bacteria was observed and their living cell absorption spectrum curve was measured. The same type of photosynthetic bacteria have similar absorption, growth, and hydrogen production characteristics. They grow faster and produce hydrogen nearer the wavelength of the maximum absorption peak. By spectral coupling characteristics for the growth and hydrogen production of photosynthetic bacteria, the hydrogen production capacity of mixed bacteria is better under the wavelength of 380–780 nm. Compared with hydrogen production under an optimal spectrum, mixed bacteria under the wavelength of 380–780 nm with the illuminance of 2000 lx has the maximum optical energy conversion rate of 13.3%, which is best for hydrogen production.

Biogenesis of photosystem II (PSII), nature’s water splitting catalyst, is assisted by auxiliary proteins that form transient complexes with PSII components to facilitate stepwise assembly events. Using cryo-electron microscopy, we solved the structure of such a PSII assembly intermediate with 2.94 A resolution. It contains three assembly factors (Psb27, Psb28, Psb34) and provides detailed insights into their molecular function. Binding of Psb28 induces large conformational changes at the PSII acceptor side, which distort the binding pocket of the mobile quinone (QB) and replace bicarbonate with glutamate as a ligand of the non-heme iron, a structural motif found in reaction centers of non-oxygenic photosynthetic bacteria. These results reveal novel mechanisms that protect PSII from damage during biogenesis until water splitting is activated. Our structure further demonstrates how the PSII active site is prepared for the incorporation of the Mn4CaO5 cluster, which performs the unique water splitting reaction. One Sentence Highlight The high-resolution Cryo-EM structure of the photosystem II assembly intermediate PSII-I reveals how nature’s water splitting catalyst is assembled, protected and prepared for photoactivation by help of the three assembly factors Psb27, Psb28 and Psb34.

The efficient mechanism of light capture by photosynthetic proteins allows for energy transfer and conversion to electrochemical energy at very low light intensities. In this work, reaction center (RC) proteins, or a core complex consisting of the RC encircled by light harvesting (LH1) proteins (RC-LH1) from photosynthetic bacteria, were immobilized on an insulating layer of an ion-sensitive field-effect transistor (ISFET) to build bio-photodetectors. The orientation of the RC proteins was controlled via application of a hybrid linker made of 10-carboxydecylphosphonic acid and cytochrome c that anchored the RCs to their electron donor side. Bio-phototransistors consisting of either the core RC or the RC-LH1 core complex were tested under white and monochromic light. The difference between the dark and light currents at different wavelengths are well-matched with the absorption spectrum of the photosynthetic proteins. The results show potential for the use of photosynthetic proteins in photodetectors.

Photosynthetic bacteria have been proven to be excellent model organisms because they own the relatively simplified model systems for us to study the reactions that occurs at the initial stage of photosynthesis, compared with the oxygen-evolving cyanobacteria, algae, and higher plants. In purple bacteria, there are usually two kinds of light-harvesting (LH) complexes, named LH1 and LH2, respectively. LH2 is the peripheral antenna complex, and LH1 is the core antenna complex that surrounds the reaction center (RC) to form the LH1-RC supercomplex. Solar energy is first absorbed by the LH complex and then transferred rapidly and efficiently to the RC, where the charge separation and electron transfer take place. Several high-resolution structures are available for the RC and LH2 for a long time; for LH1-RC complex, its structure was solved and improved to an atomic resolution recently with a thermophilic purple photosynthetic bacterium Thermochromatium tepidum. The high-resolution structure provided much more detailed structural information of this supercomplex including the arrangements of protein subunits, pigments, and cofactors; a much more intact RC complex due to the protection of LH1 complex; the detailed coordination of the Ca2+ ions in the LH1 that are important for the absorption maximum at 915 nm as well as for the enhanced thermostability; the possible ubiquinone exchange pathway in the closed LH1 ring; and so on. In addition, the dynamic processes involved in this complex were also discussed. All these results greatly advance our understanding on the molecular mechanism of bacterial photosynthesis, which could be essential for designing artificial photoelectronic conversion materials with enhanced performance.

A novel red emissive carbon dots (CDs) derived from photosynthetic bacteria are synthesized successfully via a simple microwave–assisted hydrothermal process. The CDs exhibit good water solubility, biocompatibility, and red photoluminescence emission with an absolute quantum yield of 15.17% in water. More importantly, 5 mg/ml CDs present low toxicity after co–cultured with human umbilical vein endothelial cells (HUVECs) and HeLa cells. Furthermore, the red emissive CDs can clearly image the HUVECs and HeLa cells within 10 min and maintain stable emission for 1 h. It is also proved that the organelle of CD imaging is mainly lysosome. This work provides a new method to prepare long–wavelength fluorescence–emission CDs and explores a new material with potential applications in the biomedical field.

The direct connection between a photosensitizer and an electron mediator, achieved by a precise arrangement based on chlorosome, provides photosynthetic bacteria the maximum efficiency in solar energy conversion. Herein, this study reports the fabrication of a biomimicking chlorosome for biocatalyzed artificial photosynthesis through self-assembly of simple molecules to functional systems. TCPP/EYx/Rh8–x macromolecules, synthesized through a sequential amidation reaction of porphyrin (TCPP), eosin Y (EY), and [Cp*RhCl2]2, were found to self-assemble into chlorosome-mimicking supramolecular assemblies through noncovalent interactions. Intra- and intermolecular dual channels for enhancing electron transfer were constructed in TCPP/EYx/Rh8–x supramolecular assemblies. In addition, the energy band structure of TCPP/EY4/Rh4 supramolecular assemblies also made a perfect coordination with oxidation and reduction potentials of an electron donor and NAD+, which led to a fast and oriented electron transfer along th...

Sewage lagoons and wastewater ponds from industrialised swine and poultry farms are typically hypereutrophic, auxinic and dominated by purple non-sulphur bacteria and unicellular green algae both typically growing photoheterotrophically. To manage such ponds, it is essential to know the balance between oxygenic and anoxygenic photosynthesis. Typical spectrophotometric algorithms to estimate chlorophyll use 750 nm as a zero (A750 nm) but a 750 nm zero protocol is unsuitable where substantial amounts of bacteriochlorophylls are present. Algorithms were developed to estimate chlorophylls a and b (Chl a and Chl b) and bacteriochlorophyll a (BChl a) in solvent in ethanol, 7:2 acetone/ethanol and 90% acetone. The algorithms use an 850-nm absorbance zero (A850 nm) well outside the absorbance ranges of both chlorophylls and bacteriochlorophylls in solvent. There are many habitats where the presence of anoxygenic photosynthetic bacteria is unsuspected and so using a routine A750 nm zero effectively masks their presence and leads to underestimations of Chl a and Chl b. The in-solvent red peak of bacteriochlorophyll c is too close to that of Chl a for BChl c and Chl a to be resolved spectroscopically, but its presence can be easily identified from in vivo scans. The spectroscopic advantage of 90% acetone is negated by its poor quantitative extraction of pigments. Acetone/ethanol (7:2) is an excellent solvent spectroscopically and as an extractant.

The kinetics of bacteriochlorophyll fluorescence in intact cells of the purple nonsulfur bacterium Rhodobacter sphaeroides were measured under continuous and pulsed actinic laser diode (808 nm wavelength and maximum 2 W light power) illumination on the micro- and millisecond timescale. The fluorescence induction curve was interpreted in terms of a combination of photochemical and triplet fluorescence quenchers and was demonstrated to be a reflection of redox changes and electron carrier dynamics. By adjustment of the conditions of single and multiple turnovers of the reaction center, we obtained 11 ms–1 and 120 μs–1 for the rate constants of cytochrome c23+ detachment and cyclic electron flow, respectively. The effects of cytochrome c2 deletion and chemical treatments of the bacteria and the advantages of the fluorescence induction study on the operation of the electron transport chain in vivo were discussed.

Green photosynthetic bacteria have extramembranous light-harvesting complexes called chlorosomes. Light-energy captured by bacteriochlorophyll (BChl) self-aggregates inside chlorosomes is transferred to BChl a -peptide complexes called baseplates on the chlorosome surface. We substitute the baseplate complex in chlorosomes from a green sulfur photosynthetic bacterium Chlorobaculum limnaeum with amphiphilic Zn BChl a derivatives. Zn BChl a derivatives 1 – 3 , which were esterified with polyethylene glycol (PEG) 600, PEG400, and diethylene glycol, respectively, were successfully hybridized with chlorosomes without baseplates. These derivatives in the hybridized chlorosomes emitted by predominant excitation of BChl e self-aggregates at 460 nm, indicating that they were able to accept the excitation energy from BChl e aggregates inside chlorosomes. Zn BChl a derivatives 1 and 2 emitted at 787 nm by the excitation energy transfer. In contrast, fluorescence from 3 esterified with a short hydrophilic chain, diethylene glycol, was broadened around 785 and 820 nm. The broad emission bands would consist of at least two bands, which were ascribable to the monomeric and aggregated forms. These results indicate that PEG600 and PEG400 provide appropriate amphiphilicity to Zn BChl a derivatives for dispersion in the membranous region of chlorosomes. The present study will be helpful for not only elucidation of the energy transfer mechanism in chlorosomes but also development of artificial photofunctional supramolecular complexes by hybridization of natural photosynthetic systems with synthetic molecules.

Self-aggregates of zinc 31-hydroxy-chlorins were prepared in a polydimethylsiloxane (PDMS) oil. The artificial self-aggregates provided a similar chlorophyllous aggregate found in light-harvesting antennas of green photosynthetic bacteria. The zinc chlorin aggregates showed a red-shifted Qy absorption band as well as intense circular dichroism signals at room temperature due to strong exciton coupling among the pigment molecules along their molecular y-axes. Disaggregation studies of the zinc chlorin aggregates were successfully performed in PDMS at a high temperature, which indicated that the aggregate of the 31R-stereoisomer of zinc 3-(1-hydroxyethyl)-chlorin was more stable than that of the corresponding 31S-isomer. In addition, mixed aggregates of the 31R- and 31S-epimers prepared in PDMS provided a good structural model of the natural light-harvesting system.

Mercuric contamination of aqueous cultures results in impairment of viability of photosynthetic bacteria primarily by inhibition of the photochemistry of the reaction center (RC) protein. Isolated reaction centers (RCs) from Rhodobacter sphaeroides were exposed to Hg2+ ions up to saturation concentration (~ 103 [Hg2+]/[RC]) and the gradual time- and concentration-dependent loss of the photochemical activity was monitored. The vast majority of Hg2+ ions (about 500 [Hg2+]/[RC]) had low affinity for the RC [binding constant Kb ~ 5 mM−1] and only a few (~ 1 [Hg2+]/[RC]) exhibited strong binding (Kb ~ 50 μM−1). Neither type of binding site had specific and harmful effects on the photochemistry of the RC. The primary charge separation was preserved even at saturation mercury(II) concentration, but essential further steps of stabilization and utilization were blocked already in the 5 < [Hg2+]/[RC]

In almost all photosynthetic organisms the photosynthetic pigments chlorophyll and bacteriochlorophyll (BChl) are Mg2+ containing complexes, but Mg2+ may be exchanged against other metal ions when these are present in toxic concentrations, leading to inactivation of photosynthesis. In this report we studied mechanisms of copper toxicity to the photosynthetic apparatus of Acidiphilium rubrum, an acidophilic purple bacterium that uses Zn2+ instead of Mg2+ as the central metal in the BChl molecules ([Zn]-BChl) of its reaction centres (RCs) and light harvesting proteins (LH1). We used a combination of in vivo measurements of photosynthetic activity (fast fluorescence and absorption kinetics) together with analysis of metal binding to pigments and pigment-protein complexes by HPLC-ICP-sfMS to monitor the effect of Cu2+ on photosynthesis of A. rubrum. Further, we found that its cytoplasmic pH is neutral. We compared these results with those obtained from Rhodospirillum rubrum, a purple bacterium for which we previously reported that the central Mg2+ of BChl can be replaced in vivo in the RCs by Cu2+ under environmentally realistic Cu2+ concentrations, leading to a strong inhibition of photosynthesis. Thus, we observed that A. rubrum is much more resistant to copper toxicity than R. rubrum. Only slight changes of photosynthetic parameters were observed in A. rubrum at copper concentrations that were severely inhibitory in R. rubrum and in A. rubrum no copper complexes of BChl were found. Altogether, the data suggest that [Zn]-BChl protects the photosynthetic apparatus of A. rubrum from detrimental insertion of Cu2+ (trans-metallation) into BChl molecules of its RCs.

Chlorosomes are the main light-harvesting complexes of green photosynthetic bacteria that are adapted to a phototrophic life at low-light conditions. They contain a large number of bacteriochlorophyll c, d, or e molecules organized in self-assembling aggregates. Tight packing of the pigments results in strong excitonic interactions between the monomers, which leads to a redshift of the absorption spectra and excitation delocalization. Due to the large amount of disorder present in chlorosomes, the extent of delocalization is limited and further decreases in time after excitation. In this work we address the question whether the excitonic interactions between the bacteriochlorophyll c molecules are strong enough to maintain some extent of delocalization even after exciton relaxation. That would manifest itself by collective spontaneous emission, so-called superradiance. We show that despite a very low fluorescence quantum yield and short excited state lifetime, both caused by the aggregation, chlorosomes indeed exhibit superradiance. The emission occurs from states delocalized over at least two molecules. In other words, the dipole strength of the emissive states is larger than for a bacteriochlorophyll c monomer. This represents an important functional mechanism increasing the probability of excitation energy transfer that is vital at low-light conditions. Similar behaviour was observed also in one type of artificial aggregates, and this may be beneficial for their potential use in artificial photosynthesis.

Upon high light excitation in photosynthetic bacteria, various triplet states of pigments can accumulate leading to harmful effects. Here, the generation and lifetime of flash-induced carotenoid triplets (3Car) have been studied by observation of the quenching of bacteriochlorophyll (BChl) fluorescence in different strains of photosynthetic bacteria including Rvx. gelatinosus (anaerobic and semianaerobic), Rsp. rubrum, Thio. roseopersicina, Rba. sphaeroides 2.4.1 and carotenoid- and cytochrome-deficient mutants Rba. sphaeroides Ga, R-26, and cycA, respectively. The following results were obtained: (1) 3Car quenching is observed during and not exclusively after the photochemical rise of the fluorescence yield of BChl indicating that the charge separation in the reaction center (RC) and the carotenoid triplet formation are not consecutive but parallel processes. (2) The photoprotective function of 3Car is not limited to the RC only and can be described by a model in which the carotenoids are distributed in the lake of the BChl pigments. (3) The observed lifetime of 3Car in intact cells is the weighted average of the lifetimes of the carotenoids with various numbers of conjugated double bonds in the bacterial strain. (4) The lifetime of 3Car measured in the light is significantly shorter (1–2 μs) than that measured in the dark (2–10 μs). The difference reveals the importance of the dynamics of 3Car before relaxation. The results will be discussed not only in terms of energy levels of the 3Car but also in terms of the kinetics of transitions among different sublevels in the excited triplet state of the carotenoid.

In reaction centers (RCs) of photosynthesis, a light-induced charge separation takes place creating radical cations and anions of the participating cofactors. In photosynthetic bacteria, different bacteriochlorophylls (BChl) are involved in this process. Information about the electronic structure of the BChl radical cations and anions can be obtained by measuring the electron spin density distribution via the electron-nuclear hyperfine interaction using EPR and ENDOR techniques. In this communication, we report isotropic hyperfine coupling constants (hfcs) of the BChl b and g radical cations and anions, calculated by density functional theory, and compare them with the more common radical ions of BChl a and with available experimental data. The observed differences in the computed hyperfine data are discussed in view of a possible distinction between these species by EPR/ENDOR methods. In addition, 14 N nuclear quadrupole coupling constants (nqcs) computed for BChl a, b, g, and also for Chl a in their charge neutral, radical cation and radical anion states are presented. These nqcs are compared with experimental values obtained by ESEEM spectroscopy on several different radical ions.