Light-induced Electron Transport Research Articles

A green and easy-to-assemble electrochemical biosensor based on thylakoid membranes for photosynthetic herbicides detection

In this study, we report on an easy-to-assemble amperometric electrochemical biosensor incorporating thylakoid membranes for the detection of photosynthetic herbicides. These molecules interfere with the light-induced photosynthetic electron transport occurring at the level of the photosystems within the thylakoid membranes, thus reducing the current of the associated bioelectrode. Thylakoid membranes isolated from pea plants were adsorbed directly on a bare carbon paper working electrode and placed in the measurement cell in the absence of any electrochemical mediator, obtaining a fully environmental-friendly biodevice capable of photocurrent densities up to 14 μA/cm2. Three photosynthetic herbicides inhibiting Photosystem II and belonging to different chemical classes, namely diuron, terbuthylazine and metribuzin, were detected by measuring the electrode photocurrent, which decreased reproducibly in a concentration-dependent manner in a range between 10−7 – 5 × 10−5 M of each herbicide. The limit of detection for the three herbicides was between 4–6 × 10−7 M. Storage stability tests revealed for the biosensor a half-life longer than 15 days at 4 °C and full stability up to 4 months at −80 °C. This study provides a simple, environmental-friendly and cost-effective procedure for the fabrication of a mediatorless carbon paper-based electrochemical biosensor characterized by high photocurrents, long storage stability, reproducible detections and good sensitivity.

Chloroplast-containing coacervate micro-droplets as a step towards photosynthetically active membrane-free protocells.

Encapsulation of structurally and functionally intact chloroplasts within coacervate micro-droplets is used to prepare membrane-free protocells capable of light-induced electron transport.

Electron Transfer through the Acceptor Side of Photosystem I: Interaction with Exogenous Acceptors and Molecular Oxygen.

This review considers the state-of-the-art on mechanisms and alternative pathways of electron transfer in photosynthetic electron transport chains of chloroplasts and cyanobacteria. The mechanisms of electron transport control between photosystems (PS) I and II and the Calvin-Benson cycle are considered. The redistribution of electron fluxes between the noncyclic, cyclic, and pseudocyclic pathways plays an important role in the regulation of photosynthesis. Mathematical modeling of light-induced electron transport processes is considered. Particular attention is given to the electron transfer reactions on the acceptor side of PS I and to interactions of PS I with exogenous acceptors, including molecular oxygen. A kinetic model of PS I and its interaction with exogenous electron acceptors has been developed. This model is based on experimental kinetics of charge recombination in isolated PS I. Kinetic and thermodynamic parameters of the electron transfer reactions in PS I are scrutinized. The free energies of electron transfer between quinone acceptors A1A/A1B in the symmetric redox cofactor branches of PS I and iron-sulfur clusters FX, FA, and FB have been estimated. The second-order rate constants of electron transfer from PS I to external acceptors have been determined. The data suggest that byproduct formation of superoxide radical in PS I due to the reduction of molecular oxygen in the A1 site (Mehler reaction) can exceed 0.3% of the total electron flux in PS I.

Structural Design Principle of Small-Molecule Organic Semiconductors for Metal-Free, Visible-Light-Promoted Photocatalysis.

Herein, we report on the structural design principle of small-molecule organic semiconductors as metal-free, pure organic and visible light-active photocatalysts. Two series of electron-donor and acceptor-type organic semiconductor molecules were synthesized to meet crucial requirements, such as 1) absorption range in the visible region, 2) sufficient photoredox potential, and 3) long lifetime of photogenerated excitons. The photocatalytic activity was demonstrated in the intermolecular C-H functionalization of electron-rich heteroaromates with malonate derivatives. A mechanistic study of the light-induced electron transport between the organic photocatalyst, substrate, and the sacrificial agent are described. With their tunable absorption range and defined energy-band structure, the small-molecule organic semiconductors could offer a new class of metal-free and visible light-active photocatalysts for chemical reactions.

Plasmon-Mediated Electron Transport in Tip-Enhanced Raman Spectroscopic Junctions.

We combine experiment, theory, and first-principles-based calculations to study the light-induced plasmon-mediated electron transport characteristics of a molecular-scale junction. The experimental data show a nonlinear increase in electronic current perturbation when the focus of a chopped laser beam moves laterally toward the tip-sample junction. To understand this behavior and generalize it, we apply a combined theory of the electronic nonequilibrium formed upon decoherence of an optically triggered plasmon and first-principles transport calculations. Our model illustrates that the current via an adsorbed molecular monolayer increases nonlinearly as more energy is pumped into the junction due to the increasing availability of virtual molecular orbital channels for transport with higher injection energies. Our results thus illustrate light-triggered, plasmon-enhanced tunneling current in the presence of a molecular linker.

Characteristics of light-induced electron transport from P3HT to ZnO-nanowire field-effect transistors

We fabricated ZnO-nanowire (NW) field-effect transistors (FETs) coated with poly(3-hexylthiophene) (P3HT) and characterized the electron-transfer characteristics from the P3HT to the ZnO NWs. Under irradiation by laser light with a wavelength of 532 nm, photo-induced electrons were created in the P3HT and then transported to the ZnO NWs, constituting a source-drain current in the initially enhancement-mode P3HT-coated ZnO-NW FETs. As the intensity of the light increased, the current increased, and its threshold voltage shifted to the negative gate-bias direction. We estimated the photo-induced electron density and the electron-transfer characteristics, which will be helpful for understanding organic-inorganic hybrid optoelectronic devices.

Rapid degradation of the tetrameric Mn cluster in illuminated, PsbO-depleted photosystem II preparations

A "decoupling effect" (light-induced electron transport without O2 evolution) was observed in Ca-depleted photosystem II (PSII(-Ca)) membranes, which lack PsbP and PsbQ (Semin et al. (2008) Photosynth. Res., 98, 235-249). Here PsbO-depleted PSII (PSII(-PsbO)) membranes (which also lack PsbP and PsbQ) were used to examine effects of PsbO on the decoupling. PSII(-PsbO) membranes do not reduce the acceptor 2,6-dichlorophenolindophenol (DCIP), in contrast to PSII(-Ca) membranes. To understand why DCIP reduction is lost, we studied light effects on the Mn content of PSII(-PsbO) samples and found that when they are first illuminated, Mn cations are rapidly released from the Mn cluster. Addition of an electron acceptor to PSII(-PsbO) samples accelerates the process. No effect of light was found on the Mn cluster in PSII(-Ca) membranes. Our results demonstrate that: (a) the oxidant, which directly oxidizes an as yet undefined substrate in PSII(-Ca) membranes, is the Mn cluster (not the Y(Z) radical or P680(+)); (b) light causes rapid release of Mn cations from the Mn cluster in PSII(-PsbO) membranes, and the mechanism is discussed; and (c) rapid degradation of the Mn cluster under illumination is significant for understanding the lack of functional activity in some PSII(-PsbO) samples reported by others.

Kinetics of in vivo bacteriochlorophyll fluorescence yield and the state of photosynthetic apparatus of purple bacteria

The light-induced electron transport in purple bacterium Rhodobacter sphaeroides was studied in vivo by means of kinetic difference absorption spectroscopy and kinetics of bacteriochlorophyll fluorescence yield. Measurements of redox state of the oxidised primary donor and cytochrome c and the membrane potential revealed a complex pattern of changes of the electron flow. Effects of the membrane potential on the fluorescence yield were also analysed, and a model for the fluorescence induction curve is presented. The data indicate substantial positive effect of the membrane potential on the fluorescence emission in vivo. Moreover, light-induced changes in light scattering were observed, which suggests occurrence of structural changes on the level of the photosynthetic membrane.

The Occurrence of the psbS Gene Product in Chlamydomonas reinhardtii and in Other Photosynthetic Organisms and Its Correlation with Energy Quenching†

To avoid photodamage, photosynthetic organisms have developed mechanisms to evade or dissipate excess energy. Lumen overacidification caused by light-induced electron transport triggers quenching of excited chlorophylls and dissipation of excess energy into heat. In higher plants participation of the PsbS protein as the sensor of low lumenal pH was clearly demonstrated. Although light-dependent energy quenching is a property of all photosynthetic organisms, large differences in amplitude and kinetics can be observed thus raising the question whether a single common mechanism is in action. We performed a detailed study of PsbS expression/accumulation in Chlamydomonas reinhardtii and investigated its accumulation in other algae and plants. We showed that PsbS cannot be detected in Chlamydomonas under a wide range of growth conditions. Overexpression of the endogenous psbs gene showed that the corresponding protein could not be addressed to the thylakoid membranes. Survey of different unicellular green algae showed no accumulation of anti-PsbS reactive proteins differently from multicellular species. Nevertheless, some unicellular species exhibit high energy quenching activity, suggesting that a PsbS-independent mechanism is activated. By correlating growth habitat and PsbS accumulation in different species, we suggest that during the evolution the light environment has been a determinant factor for the conservation/loss of the PsbS function.

Effects of Dark Adaptation on Light-Induced Electron Transport through Photosystem I in the Cyanobacterium Synechocystis sp. PCC 6803

Kinetics of the redox reactions in the reaction center (P700) of photosystem I (PSI) of the cyanobacterium Synechocystis sp. PCC 6803 have been studied by EPR spectroscopy. The redox kinetics were recorded based on accumulation of the EPRI signal when the final signal was the sum of individual signals produced in response to illumination of the cells. After prolonged (more than 3 sec) dark intervals between illuminations, the kinetic curve of the EPR signal from P700+ was multiphasic. After a sharp increase in the signal amplitude at the beginning of illumination (phase I), the amplitude rapidly (for 0.1-0.2 sec) decreased (phase II). Then the signal amplitude gradually increased (phase III) until the steady rate of electron transfer was established. With short-term (1 sec) dark intervals between the flashes and also in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), the kinetics of the light-induced increase in the EPR signal from P700+ were monophasic. Inhibition with iodoacetamide of electron transport on the acceptor side of PSI under anaerobic conditions or an increase in the amount of respiration substrates on addition of glucose into a suspension of DCMU-treated wild-type cells increased the level of P700 reduction in phase III. The findings suggest that the kinetic curve of the EPR signal from P700+ is determined by both the electron entrance onto P700+ on the donor side of PSI and activity of electron acceptors of PSI.

Biological and polymeric self-assembled hybrid systems: Structure and properties of thylakoid/polyelectrolyte complexes

A novel hybrid system composed of biological components and synthetic polymer, thylakoid/polycation complex, has been formed and studied. Effects of complex formation on the structure, electrostatics and functioning of thylakoid membranes have been examined. Thylakoids from bean leaves were used to form complexes with polycation polyallylamine hydrochloride (PAAH) in two systems: (i) thylakoid/polycation complexes formed in an aqueous bulk phase, and (ii) immobilized thylakoid/polycation planar complexes. Immobilized on a solid substrate surface, thylakoid/polycation complexes were prepared using layer-by-layer stepwise alternate adsorption technique, i.e., via the sequential alternate adsorption of thylakoids and polycation molecules. The morphology of built up structures was investigated by scanning electron microscopy. Light-induced electron transport in chloroplasts was studied by the electron paramagnetic resonance (EPR) method. Spin probe technique was employed to study the structural and electrostatic characteristics of thylakoid membranes. We have found that efficiency of light-induced electron transport in thylakoid membranes and membrane structure were not changed noticeably by PAAH binding to thylakoids in a wide range of PAAH concentrations. The data obtained indicate the physiologically-soft character of polycation interactions with thylakoid membranes and demonstrate effectiveness of interfacial self-assembly approach to fabrication of complex planar functional nanostructures from biological components and synthetic polymers.

A biocatalyst for the removal of sulfite from alcoholic beverages

The presence of sulfites in alcoholic beverages, particularly in wines, can cause allergic responses with symptoms ranging from mild gastrointestinal problems to life threatening anaphylactic shock in a substantial portion of the population. We have developed a simple and inexpensive biocatalytic method that employs wheatgrass (Triticum aestivum) chloroplasts for the efficient oxidation of sulfites in wines to innocuous sulfates. A sufficiently high rate of sulfite oxidation was obtained in the presence of ethanol at concentrations commonly found in most wines. Crude chloroplast preparations at a concentration as low as 5 mg/mL were capable of reducing sulfite in commercial white wines from 150 ppm to under 7.5 ppm within 3 hours. A 93% removal of sulfite in commercial red wines was observed with 1 mg/mL chloroplasts within 45 min. Optimal sulfite removal efficiency was observed at pH 8.5 and was promoted by illumination, indicating the participation of light-induced photosynthetic electron transport processes in sulfite oxidation. Overall, this work indicates that biocatalytic oxidation using wheatgrass chloroplasts can be employed to remove sulfites from beverages prior to consumption.

Photosystem I reaction center: past and future.

Science has always been drawn to uncover fundamental life processes. Photosynthesis is one, if not the most fascinating, of them. Within it, the protein complexes that catalyze light-induced electron transport and photophosphorylation are enchanting creations of evolution. Plant Photosystem I (PS I) is not the largest protein complex in nature but it is the most elaborate in the number of prosthetic groups involved in its fabric. Thirty years ago, one of us (NN) developed a fascination for this complex and, despite the apparent neglect (lack of publications in the last few years), never let it go. Only a crystal structure at 2 A resolution will satiate our curiosity. In this minireview, we trace the past, and end the article with a comment on future prospects. For the present situation, see Parag Chitnis (2001).

Light-Induced Ascorbate-Dependent Electron Transport and Membrane Energization in Chloroplasts of Bundle Sheath Cells of the C4 Plant Maize

Chloroplasts in bundle sheath cells (BSC) of maize perform photosystem I (PSI)-mediated production of ATP. In this study, the participation of ascorbate (Asc) as an electron donor to PSI in light-induced electron transport in isolated maize BSC was demonstrated. It was found that Asc, at physiological concentrations, rapidly reduced photooxidized reaction center chlorophyll of PSI (P700). The rate of Asc donation of electrons to P700+ reached rates of 50–100 μequivalents (mg Chl)−1 h−1 at 70–80 mM ascorbate with methyl viologen as an electron acceptor. Electron transport supported by Asc was coupled with membrane energization, as demonstrated by the light-induced formation of a trans-thylakoid electric field measured by the electrochromic shift of carotenoids. The possible physiological function of Asc-dependent electron transport in bundle sheath chloroplasts of maize, as an electron donor for linear electron flow versus sustaining cyclic electron transport, is discussed.

Allelopathy effects of aqueous rinses of Dittrichia viscosa (L.) on the photosynthesis and cell proliferation of N2-fixing soil cyanobacteriaK Stamatakis1, M Konstantopoulou1Institute of Biology, NCSR Demokritos, Aghia Paraskevi, 15310, Athens,

Dittrichia viscosa (L.) W. Greuter (syn. Inula viscosa (L.) Aiton) (Asteraceae) is an evergreen shrub, widespread in the Mediterranean region. Leaf surface of D. viscosa is covered with an epicuticular material that contains numerous terpenoids and phenolic compounds. Aqueous rinses of leaves of D. viscosa showed strong allelopathic effects against a spectrum of different organisms, including plants and pathogens. Rain rinses of D. viscosa end up in the soil. In the present study we investigated the allelopathy effects of the epicuticular rinses from D. viscosa on growth, photosynthetic activity, respiration and heterosyst formation of the N2-fixing soil cyanobacteria Nostoc sp. PCC 7413 and Anabaena sp. PCC 6309. Epicuticular substances from leaves of D. viscosa inhibited cell proliferation of these cyanobacteria in a dose-related manner. These substanses, however, exerted no effect on light-induced photosynthetic electron transport (from water to ferredoxin). This indicates inhibitory action on dark metabolic processes, such as CO2 fixation and respiration. We observed also increased heterocyst formation in the presence of the epicuticular material. This work was supported by a grant (99ED 121) from the Greek Ministry of Industry, Energy and Technology and the European Social Fund.

Nonphotosynthetic Reduction of the Intersystem Electron Transport Chain of Chloroplasts Following Heat Stress. Steady-state Rate

The consequence of elevated temperatures in the range of 39-51 degrees C on the steady-state rate of light-induced electron transport through photosystem I (PSI) supported by stromal reductants was studied in intact barley leaves using photoacoustic and chlorophyll fluorescence techniques. Measurable electron flow through PSI in diuron-treated leaves occurred only after exposure to temperatures above 37 degrees C. The steady-state rate of the above diuron-insensitive electron flow with methyl viologen as electron acceptor was estimated to be 3.7 mu eq m-2 s-1 or 0.018 mu eq mumol chlorophyll-1 s-1 in leaves exposed for 5 min to 45 degrees C.

Two distinct states of the thylakoid bf complex

Under normal physiological conditions the state of the cyt bf complex is characterized by rapid reoxidation kinetics of cyt b-563 following flash-illumination. It is known that these kinetics are dramatically slowed down under oxidizing conditions. Here we show that this slow-down of cyt b-563 oxidation is the consequence of a relatively slow (half-time of several minutes) transformation of the cyt bf complex into a distinctly different state (termed state-s). Reversal to the normal state requires strong reductive treatment or light-induced electron transport. The results are in line with a recent model of functional cyt bf dimers [Cramer et al., Annu. Rev. Plant Physiol. Plant Mol. Biol. 47 (1996), 477–508], if it is assumed that state-s reflects the monomeric state of the bf complex.

Energy and entropy balance of ATP synthesis

Analysis of the energy balance at the H +-translocating CF 1CF 0–ATP synthase from spinach yields precise data for the H +/ATP coupling ratio and the standard reaction Gibbs energy, Δ r G′ 0, of the reaction ADP+P i→ATP+H 2O. From the temperature dependency of Δ r G′ 0 the accompanying Δ r H′ 0 and Δ r S′ 0 values are derived. Prerequisite of this analysis is a shift of the reaction quotient from values far below 1 for the uncoupled reaction to values near 1 by means of the coupling to the energetically downhill translocation of protons across the thylakoid membrane. Proton load of the thylakoid lumen is achieved along with the light-induced electron transport. The internal pH which is of crucial importance for the energy balance analysis is obtained from the pH in-controlled electron flow rate. Linear relationships between the logarithm of the reaction quotient and the equilibrium transmembrane pH difference prove the reliability of the procedure. The quantitative results are: (1) H +/ATP=4.0, independent of pH. (2) Δ r G′ 0=31.3 kJ mol −1 at conditions of 20°C, pH 8.0, pMg 2.5 and 0.08 M ionic strength. (3) Δ r H′ 0=28.1 kJ mol −1, Δ r S′ 0=−11 J K −1 mol −1. These results are discussed with regard to the nonuniform data in the literature.

Donor function of phosphate ions in photosystem 2

Evidence was obtained for the interaction between the photosystem 2 (PS2) reaction centre (RC) chlorophyll (Chl) P680 and inorganic phosphate, Pi. The light-induced endogenous basal electron transport to ferricyanide in PS2 depended on endogenous Pi. The electron transport in phosphate deficient was absent, and could be resumed upon the addition of exogenous Pi or of the exogenous electron donor, diphenylcarbazide. Some chloroplast Chl molecules were apparently bound with Pi to a complex via the magnesium atom that was detected by the increase in absorbance in the Chl a absorption maximum at 435 nm observed after the consumption of endogenous Pi in the photophosphorylation reactions. The electron paramagnetic resonance (EPR) Signal I, found in the spectra at 77 K after irradiation of frozen samples in poor in endogenous Pi, was the sum of P700+ and P680+ signals. The P680+ signal disappeared after addition of Pi, diphenylcarbazide or to the before freezing. In addition, the EPR doublet signal of the phosphate anion radicals was recorded at 77 K after irradiation in the ethanol solutions of Chl a containing potassium phosphate. The same doublet signal was discovered in the difference EPR spectrum chloroplasts minus with diuron at 77 K after irradation. The results are a possible evidence of the participation of phosphate ions in the primary light reactions of PS2.

Membrane vesicles fromSynechocystis 6803 showing proton and electron transport and high ATP synthase activities

A simple procedure for the preparation of well-coupled and stable membrane vesicles from the transformable cyanobacteriumSynechocystis 6803 is described with the primary aim of producing vesicles suitable for the study of photosynthetic electron transport and phosphorylation. Spheroplasts were obtained from the cyanobacterium by lysozyme treatment and stored untill prior to measurement, thylakoid vesicles were obtained by osmotic shock. These vesicles showed very high and stable ATP synthesis rates either driven by light or by acid-base transition, and also performed light-induced ATP hydrolysis and linear electron transport. Formation of a proton gradient is studied by aminoacridines.