Photo-induced processes in photosynthesis-from femtoseconds to seconds.

Photosynthesis nourishes nearly all life on Earth. Therefore, a deeper understanding of the processes by which sunlight is converted into stored chemical energy presents an important and continuing challenge for fundamental scientific research. This Special Issue is dedicated to academician Vladimir A. Shuvalov (1943-2022). We are delighted to present 15 manuscripts in the Special Issue, including two review articles and 13 research papers. These papers are contributed by 67 authors from 8 countries, including China (9), Germany (8), Hungary (4), Italy (6), Japan (2), Russia (24), Taiwan (9), and USA (5). This Special Issue provides some of the recent updates on the dynamical aspects of the initial steps of photosynthesis, including excitation energy transfer, electron transport, and dissipation of energy across time domains from femtoseconds to seconds. We hope that the readers will benefit from the work presented in this Special Issue in honor of Prof. Shuvalov in many ways. We hope that the Special Issue will provide a valued resource to stimulate research efforts, initiate potential collaboration, and promote new directions in the photosynthesis community.

Revelations on photosystem II, thermoluminescence, and artificial photosynthesis: a retrospective of Govindjee from fundamentals to applications.

Photosynthesis, as one of the most important chemical reactions, has powered our planet for over four billion years on a massive scale. This review summarizes and highlights the major contributions of Govindjee from fundamentals to applications in photosynthesis. His research included primary photochemistry measurements, in the picosecond time scale, in both Photosystem I and II and electron transport leading to NADP reduction, using two light reactions. He was the first to suggest the existence of P680, the reaction center of PSII, and to prove that it was not an artefact of Chlorophyll a fluorescence. For most photobiologists, Govindjee is best known for successfully exploiting Chlorophyll a fluorescence to understand the various steps in photosynthesis as well as to predict plant productivity. His contribution in resolving the controversy on minimum number of quanta in favor of 8-12 vs 3-4, needed for the evolution of one molecule of oxygen, is a milestone in the area of photosynthesis research. Furthermore, together with Don DeVault, he is the first to provide the correct theory of thermoluminescence in photosynthetic systems. His research productivity is very high: ~ 600 published articles and total citations above 27,000 with an h-index of 82. He is a recipient of numerous awards and honors including a 2022: Lifetime Achievement Award of the International Society of Photosynthesis Research. We hope that the retrospective of Govindjee described in this work will inspire and stimulate the readers to continue probing the photosynthetic apparatuses with new discoveries and breakthroughs.

Polyphosphate kinase deletion increases laboratory productivity in cyanobacteria.

Identification and manipulation of cellular energy regulation mechanisms may be a strategy to increase productivity in photosynthetic organisms. This work tests the hypothesis that polyphosphate synthesis and degradation play a role in energy management by storing or dissipating energy in the form of ATP. A polyphosphate kinase (ppk) knock-out strain unable to synthesize polyphosphate was generated in the cyanobacterium Synechocystis sp. PCC 6803. This mutant strain demonstrated higher ATP levels and faster growth than the wildtype strain in high-carbon conditions and had a growth defect under multiple stress conditions. In a strain that combined ppk deletion with heterologous expression of ethylene-forming enzyme, higher ethylene productivity was observed than in the wildtype background. These results support the role of polyphosphate synthesis and degradation as an energy regulation mechanism and suggest that such mechanisms may be effective targets in biocontainment design.

Bimodal intramolecular excitation energy transfer in a multichromophore photosynthetic model system: hybrid fusion proteins comprising natural phycobilin- and artificial chlorophyll-binding domains.

The phycobilisomes of cyanobacteria and red-algae are highly efficient peripheral light-harvesting complexes that capture and transfer light energy in a cascade of excitation energy transfer steps through multiple phycobilin chromophores to the chlorophylls of core photosystems. In this work, we focus on the last step of this process by constructing simple functional analogs of natural phycobilisome-photosystem complexes that are based on bichromophoric protein complexes comprising a phycobilin- and a chlorophyll- or porphyrin-binding domain. The former is based on ApcE(1-240), the N-terminal chromophore-binding domain of the phycobilisome's L(CM) core-membrane linker, and the latter on HP7, a de novo designed four-helix bundle protein that was originally planned as a high-affinity heme-binding protein, analogous to b-type cytochromes. We fused a modified HP7 protein sequence to ApcEΔ, a water-soluble fragment of ApcE(1-240) obtained by excising a putative hydrophobic loop sequence of residues 77-153. HP7 was fused either to the N- or the C-terminus of ApcEΔ or inserted between residues 76 and 78, thereby replacing the native hydrophobic loop domain. We describe the assembly, spectral characteristics, and intramolecular excitation energy transfer of two unique systems: in the first, the short-wavelength absorbing zinc-mesoporphyrin is bound to the HP7 domain and serves as an excitation-energy donor to the long-wavelength absorbing phycocyanobilin bound to the ApcE domain; in the second, the short-wavelength absorbing phycoerythrobilin is bound to the ApcE domain and serves as an excitation energy donor to the long-wavelength absorbing zinc-bacteriochlorophyllide bound to the HP7 domain. All the systems that were constructed and tested exhibited significant intramolecular fluorescence resonance energy transfer with yields ranging from 21% to 50%. This confirms that our modular, covalent approach for studying EET between the cyclic and open chain tetrapyrroles is reasonable, and may be extended to larger structures mimicking light-harvesting in cyanobacteria. The design, construction, and characterization process demonstrated many of the advances in constructing such model systems, particularly in our ability to control the fold and aggregation state of protein-based systems. At the same time, it underlines the potential of exploiting the versatility and flexibility of protein-based systems in assembling multiple pigments into effective light-harvesting arrays and tuning the spectral properties of multichromophore systems.

Response of chlorophyll d-containing cyanobacterium Acaryochloris marina to UV and visible irradiations.

We have previously investigated the response mechanisms of photosystem II complexes from spinach to strong UV and visible irradiations (Wei et al J Photochem Photobiol B 104:118-125, 2011). In this work, we extend our study to the effects of strong light on the unusual cyanobacterium Acaryochloris marina, which is able to use chlorophyll d (Chl d) to harvest solar energy at a longer wavelength (740 nm). We found that ultraviolet (UV) or high level of visible and near-far red light is harmful to A. marina. Treatment with strong white light (1,200 µmol quanta m(-2) s(-1)) caused a parallel decrease in PSII oxygen evolution of intact cells and in extracted pigments Chl d, zeaxanthin, and α-carotene analyzed by high-performance liquid chromatography, with severe loss after 6 h. When cells were irradiated with 700 nm of light (100 µmol quanta m(-2) s(-1)) there was also bleaching of Chl d and loss of photosynthetic activity. Interestingly, UVB radiation (138 µmol quanta m(-2) s(-1)) caused a loss of photosynthetic activity without reduction in Chl d. Excess absorption of light by Chl d (visible or 700 nm) causes a reduction in photosynthesis and loss of pigments in light harvesting and photoprotection, likely by photoinhibition and inactivation of photosystem II, while inhibition of photosynthesis by UVB radiation may occur by release of Mn ion(s) in Mn4CaO5 center in photosystem II.

Methodology of pulsed photoacoustics and its application to probe photosystems and receptors.

We review recent advances in the methodology of pulsed time-resolved photoacoustics and its application to studies of photosynthetic reaction centers and membrane receptors such as the G protein-coupled receptor rhodopsin. The experimental parameters accessible to photoacoustics include molecular volume change and photoreaction enthalpy change. Light-driven volume change secondary to protein conformational changes or electrostriction is directly related to the photoreaction and thus can be a useful measurement of activity and function. The enthalpy changes of the photochemical reactions observed can be measured directly by photoacoustics. With the measurement of enthalpy change, the reaction entropy can also be calculated when free energy is known. Dissecting the free energy of a photoreaction into enthalpic and entropic components may provide critical information about photoactivation mechanisms of photosystems and photoreceptors. The potential limitations and future applications of time-resolved photoacoustics are also discussed.

Structural and mechanistic aspects of Mn-oxo and co-based compounds in water oxidation catalysis and potential applications in solar fuel production.

To address the issues of energy crisis and global warming, novel renewable carbon-free or carbon-neutral energy sources must be identified and developed. A deeper understanding of photosynthesis is the key to provide a solid foundation to facilitate this transformation. To mimic the water oxidation of photosystem II oxygen evolving complex, Mn-oxo complexes and Co-phosphate catalytic material were discovered in solar energy storage. Building on these discoveries, recent advances in solar energy conversion showed a compelling working principle by combing the active Mn-oxo and Co-based catalysts in water splitting with semiconductor hetero-nanostructures for effective solar energy harnessing. In this review the appealing systems including Mn-oxo tetramer/Nafion, Mn-oxo dimer/TiO(2), Mn-oxo oligomer/WO(3), Co-Pi/Fe(2)O(3), and Co-Pi/ZnO are summarized and discussed. These accomplishments offer a promising framework and have a profound impact in the field of solar fuel production.

Thermodynamics of charge separation of photosystem I in the menA and menB null mutants of Synechocystis sp. PCC 6803 determined by pulsed photoacoustics.

When the biosynthesis of phylloquinone is inhibited in Synechocystis sp. PCC 6803 by interrupting the menA or the menB gene, photosystem I (PS I) recruits plastoquinone-9 (A(P)) to occupy the A(1) sites. In PS I from the menA and menB null mutants, forward electron transfer from the quinone to the FeS clusters occurs approximately 1000 times slower than in wild-type PS I [Semenov, A. Yu., Vassiliev, I. R., van der Est, A., Mamedov, M. D., Zybailov, B., Shen, G., Stehlik, D., Diner, B. A., Chitnis, P. R., and Golbeck, J. H. (2000) J. Biol. Chem. 275, 23429-23438]. To investigate the effect on thermodynamics, the enthalpy and volume changes of charge separation in PS I in the menA and menB mutants were measured using pulsed time-resolved photoacoustics on the nanosecond and microsecond time scales. The observed thermodynamic data are the same for the menA and menB mutants. This is expected because the recruited quinone (A(P)) is the same in both mutants. The volume change of PS I from the mutants following charge separation on both time scales was -17 +/- 2 A(3), less than that of the wild type, -21 A(3). The quantum yield of charge separation was found to be slightly lower (85 +/- 9%) than that of wild-type PS I (96 +/- 10%). The observed reaction is assigned to the formation of P(700)(+)A(P)(-) from P(700)*A(P). An enthalpy change (DeltaH) of -0.69 +/- 0.07 eV was obtained for this reaction. In contrast, a larger enthalpy change -0.8 eV for the formation of P(700)(+)A(1)(-) from P(700)* and an apparent entropy change (TDeltaS, T = 25 degrees C) of -0.2 eV were obtained in wild-type PS I [Hou, H. J. M., and Mauzerall, D. (2006) J. Am. Chem. Soc. 128, 1580-1586]. Taking the free energy to be -0.70 eV in PS I of the mutants, the apparent entropy is close to zero in the mutants. Since the apparent entropy change for the overall reaction of the production of P(700)(+)F(A/B)(-) from P(700)* is very likely the same as that of the wild type, +0.35 eV, this implies that the reaction of P(700)(+)A(P)(-)F(A/B) --> P(700)(+)A(P)F(A/B)(-) in the mutants is almost completely entropy driven (DeltaG = -0.07 eV and TDeltaS = +0.40 eV). These results show that not only the kinetics but also the thermodynamics of electron transfer reactions in PS I are significantly affected by the recruitment of the foreign plastoquinone-9 into the A(1) site.

The Effect of Bacteriochlorophyll g Oxidation on Energy and Electron Transfer in Reaction Centers from Heliobacterium modesticaldum.

The heliobacteria are a family of strictly anaerobic, Gram-positive, photoheterotrophs in the Firmicutes. They make use of a homodimeric type I reaction center (RC) that contains ∼20 antenna bacteriochlorophyll (BChl) g molecules, a special pair of BChl g' molecules (P800), two 8(1)-OH-Chl aF molecules (A0), a [4Fe-4S] iron-sulfur cluster (FX), and a carotenoid (4,4'-diaponeurosporene). It is known that in the presence of light and oxygen BChl g is converted to a species with an absorption spectrum identical to that of Chl a. Here, we show that main product of the conversion is 8(1)-OH-Chl aF. Smaller amounts of two other oxidized Chl aF species are also produced. In the presence of light and oxygen, the kinetics of the conversion are monophasic and temperature dependent, with an activation energy of 66 ± 2 kJ mol(-1). In the presence of oxygen in the dark, the conversion occurs in two temperature-dependent kinetic phases: a slow phase followed by a fast phase with an activation energy of 53 ± 1 kJ mol(-1). The loss of BChl g' occurs at the same rate as the loss of Bchl g; hence, the special pair converts at the same rate as the antenna Chl's. However, the loss of P800 photooxidiation and flavodoxin reduction is not linear with the loss of BChl g. In anaerobic RCs, the charge recombination between P800(+) and FX(-) at 80 K is monophasic with a lifetime of 4.2 ms, but after exposure to oxygen, an additional phase with a lifetime of 0.3 ms is observed. Transient EPR data show that the line width of P800(+) increases as BChl g is converted to Chl aF and the rate of electron transfer from A0 to FX, as estimated from the net polarization generated by singlet-triplet mixing during the lifetime of P800(+)A0(-), is unchanged. The transient EPR data also show that conversion of the BChl g results in increased formation of triplet states of both BChl g and Chl aF. The nonlinear loss of P800 photooxidiation and flavodoxin reduction, the biphasic backreaction kinetics, and the increased EPR line width of P800(+) are all consistent with a model in which the BChl g'/BChl g' and BChl g'/Chl aF' special pairs are functional but the Chl aF'/Chl aF' special pair is not.

Manganese-based Materials Inspired by Photosynthesis for Water-Splitting.

In nature, the water-splitting reaction via photosynthesis driven by sunlight in plants, algae, and cyanobacteria stores the vast solar energy and provides vital oxygen to life on earth. The recent advances in elucidating the structures and functions of natural photosynthesis has provided firm framework and solid foundation in applying the knowledge to transform the carbon-based energy to renewable solar energy into our energy systems. In this review, inspired by photosynthesis robust photo water-splitting systems using manganese-containing materials including Mn-terpy dimer/titanium oxide, Mn-oxo tetramer/Nafion, and Mn-terpy oligomer/tungsten oxide, in solar fuel production are summarized and evaluated. Potential problems and future endeavors are also discussed.

Listening to PS II: enthalpy, entropy, and volume changes.

Photosystem II, located in the thylakoid membranes of green plants, algae, and cyanobacteria, uses sunlight to split water into protons, electrons, and a dioxygen molecule. The mechanism of its electron transfers and oxygen evolution including the structure of the protein and rates of the S-state cycle has been extensively investigated. Substantial progress has been made; however, the thermodynamics of PS II electron transfer and of the oxygen cycle are poorly understood. Recent progress in thermodynamic measurements in photosynthesis provides novel insights on the enthalpic and entropic contribution to electron transfer in proteins. In this review the thermodynamic parameters including quantum yield, enthalpy, entropy, and volume changes of PS II photochemistry determined by photoacoustics and other laser techniques are summarized and evaluated. Light-driven volume changes via electrostriction are directly related to the photoreaction in PS II and thus can be a useful measurement of PS II activity and function. The enthalpy changes of the reactions observed can be directly measured by photoacoustics. The apparent reaction entropy can also be estimated when the free energy is known. Dissecting the free energy of a photoreaction into enthalpic and entropic components provides critical information about mechanisms of PS II function. Potential limitations and future direction of the study of the thermodynamics of PS II electron transfer and oxygen evolution are presented.

Photodamage of a Mn(III/IV)-oxo mixed-valence compound and photosystem II: evidence that a high-valent manganese species is responsible for UV-induced photodamage of the oxygen-evolving complex in photosystem II.

The Mn cluster in photosystem II (PS II) is believed to play an important role in the UV photoinhibition of green plants, but the mechanism is still not clear at a molecular level. In this work, the photochemical stability of [Mn(III)(O)(2)Mn(IV)(H(2)O)(2)(Terpy)(2)](NO(3))(3) (Terpy=2,2':6',2''-terpyridine), designated as Mn-oxo mixed-valence dimer, a well characterized functional model of the oxygen-evolving complex in PS II, was examined in aqueous solution by exposing the complex to excess light irradiation at six different wavelengths in the range of 250 to 700 nm. The photodamage of the Mn-oxo mixed-valence dimer was confirmed by the decrease of its oxygen-evolution activity measured in the presence of the chemical oxidant oxone. Ultraviolet light irradiation induced a new absorption peak at around 400-440 nm of the Mn-oxo mixed-valence dimer. Visible light did not have the same effect on the Mn-oxo mixed-valence dimer. We speculate that the spectral change may be caused by conversion of the Mn(III)O(2)Mn(IV) dimer into a new structure--Mn(IV)O(2)Mn(IV). In the processes, the appearance of a 514 nm fluorescence peak was observed in the solution and may be linked to the hydration or protonation of Terpy ligand in the Mn-oxo dimer. In comparing the response of the PS II functional model compound and the PS II complex to excess light radiation, our results support the idea that UV photoinhibition is triggered at the Mn(4)Ca center of the oxygen-evolution complex in PS II by forming a modified structure, possibly a Mn(IV) species, and that the reaction of Mn ions is likely the initial step.

The A-Fx to F(A/B) step in synechocystis 6803 photosystem I is entropy driven.

We have previously reported the enthalpy and volume changes of charge separation in photosystem I from Synechocystis 6803 using pulsed photoacoustics on the microsecond time scale, assigned to the electron-transfer reaction from excited-state P(700) to F(A/B) iron sulfur clusters. In the present work, we focus on the thermodynamics of two steps in photosystem I: (1) P(700) --> A(1)(-)F(X) (<10 ns) and (2) A(1)(-)F(X) --> F(A/B)(-) (20-200 ns). The fit by convolution of photoacoustic waves on the nanosecond and microsecond time scales resolved two kinetic components: (1) a prompt component (<10 ns) with large negative enthalpy (-0.8 +/- 0.1 eV) and large volume change (-23 +/- 2 A(3)), which are assigned to the P(700) --> A(1)(-)F(X) step, and (2) a component with approximately 200 ns lifetime, which has a positive enthalpy (+0.4 +/- 0.2 eV) and a small volume change (-3 +/- 2 A(3)) that are attributed to the A(1)(-)F(X) --> F(A/B)(-) step. For the fast reaction using the redox potentials of A(1)F(X) (-0.67 V) and P(700) (+0.45 V) and the energy of P(700) (1.77 eV), the free energy change for the P(700) --> A(1)(-)F(X) step is -0.63 eV, and thus the entropy change (TDeltaS, T = 25 degrees C) is -0.2 +/- 0.3 eV. For the slow reaction, A(1)(-)F(X) --> F(A/B)(-), taking the free energy of -0.14 eV [Santabara, S.; Heathcote, P; Evans, C. W. Biochim. Biophys. Acta 2005, 1708, 283-310], the entropy change (TDeltaS) is positive, +0.54 +/- 0.3 eV. The positive entropy contribution is larger than the positive enthalpy, which indicates that the A(-)F(X) to F(A/B)(-) step in photosystem I is entropy driven. Other possible contributions to the measured values are discussed.