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Photosynthetic biomaterials have attracted considerable attention at different levels of the biological organisation, from molecules to the biosphere, due to a variety of artificial application possibilities. During photosynthesis, the first steps of the conversion of light energy into chemical energy take place in a pigment–protein complex, called reaction centre (RC). In our experiments photosynthetic reaction centre protein, purified from Rhodobacter sphaeroides R-26 purple bacteria, was bound to porous silicon pillars (PSiP) after the electropolymerisation of aniline onto the surface. This new type of biohybrid material showed remarkable photoactivity in terms of measured photocurrent under light excitation in an electrochemical cell. The photocurrent was found to increase considerably after the addition of ubiquinone (UQ-0), an e−-acceptor mediator of the RC. The photoactivity of the complex was found to decrease by the addition of terbutryn, the chemical which inhibits the e−-transport on the acceptor side of the RC. In addition to the generation of sizeable light-induced photocurrents, using the PSiP/RC photoactive hybrid nanocomposite material, the system was found to be sensitive towards RC inhibitors and herbicides. This highly ordered patterned 3D structure opens new solution for designing low-power (bio-)optoelectronic, biophotonic and biosensing devices. Graphical abstract

International audience; Most porous silicon-based interferometric sensors targeting biosensing applications consist of vertical porous silicon layers created into a silicon wafer by electrochemical anodization and operate in a flow-over configuration. In this work, we present an alternative porous silicon interferometer based on porous silicon with horizontally oriented pores. This architecture permits the integration of flow-through porous silicon membranes within planar microfluidics. Fourier-transform infrared spectroscopy was used to obtain interference spectra from fabricated lateral porous silicon membranes and red shifts were observed upon filling microfluidic chips integrating the porous membranes with solvents of higher optical indices. This work proves that lateral porous silicon membranes are typical Fabry-Pérot interferometers with a sensitivity of more than 150 nm/RIU and a limit of detection less than 10−3 RIU, that is comparable to vertical porous silicon layers. Moreover, we have conducted simulation studies showing that the addition of Bragg mirrors on the membranes results in spectra with narrower fringes and lateral porous silicon interferometers with improved performances. After appropriate biofunctionalization of the porous silicon surface, lateral porous silicon membrane interferometers should offer alternative solutions for the development of porous silicon flow-through biosensors monolithically integrated on-chip.

Photosynthetic biomaterials have attracted considerable attention at different levels of the biological organisation, from molecules to the biosphere, due to a variety of artificial application possibilities. During photosynthesis, the first steps of the conversion of light energy into chemical energy take place in a pigment-protein complex, called reaction centre (RC). In our experiments photosynthetic reaction centre protein, purified from Rhodobacter sphaeroides R-26 purple bacteria, was bound to porous silicon pillars (PSiP) after the electropolymerisation of aniline onto the surface. This new type of biohybrid material showed remarkable photoactivity in terms of measured photocurrent under light excitation in an electrochemical cell. The photocurrent was found to increase considerably after the addition of ubiquinone (UQ-0), an e

A parametric shrinking-core model is used to quantify the dechlorination rate of chlorinated pollutants dissolved in water by zero-valent iron nanoparticles (nZVI). The mass-transfer processes are coupled with the instantaneous reaction occurring in the solid/liquid interface to compute the transient changes of dissolved pollutant concentration in bulk and suspended nZVI concentration. A sensitivity analysis is carried out to identify the dominant parameters governing the overall reaction rate for a system of uniform nanoparticles. The model is extended to assemblages of non-uniform ZVI nanoparticles with sizes following a distribution function. The statistical shrinking-core model is used to estimate the PCE dechlorination kinetic parameters along with the mass-transfer coefficients with inverse modeling of batch experiments. The observed discrepancy between numerical predictions and experimental datasets may be attributed to uncertainties associated with the nZVI oxidation by other species, and PCE adsorbed on oxidized iron. An approximate analysis of model equations allows the development of an analytic phenomenological model providing the overall reaction rate as a function of dissolved PCE concentration.

Application of technical developments in biology and vice versa or biological samples in technology led to the development of new types of functional, so-called “biohybrid” materials. These types of materials can be created at any level of the biological organization from molecules through tissues and organs to the individuals. Macromolecules and/or molecular complexes, membranes in biology, are inherently good representatives of nanosystems since they fall in the range usually called “nano.” Nanohybrid materials provide the possibility to create functional bionanohybrid complexes which also led to new discipline called “nanobionics” in the literature and are considered as materials for the future. In this publication, the special characteristics of photosynthetic reaction center proteins, which are “nature’s solar batteries,” will be discussed in terms of their possible applications for creating functional molecular biohybrid materials.

A variety of aqueous suspensions (nanofluids) of zero-valent nano-particles (nZVI) are prepared by wet chemistry techniques, their stability and longevity is evaluated by physic-chemical methods of characterization, and their reactivity toward the dechlorination of per-chloro-ethylene (PCE) is examined with tests in batch reactors. For assessing the mobility, longevity and reactivity of nZVI suspensions (nanofluids), under flow-through conditions, visualization multiphase flow and transport tests are performed on a glass-etched pore network. The nZVI breakthrough curves are constructed by measuring the transient variation of the iron concentration in the effluent with atomic absorption spectroscopy. The capacity of nZVI to remediate the bulk phase of PCE is quantified by detecting the mass loss rate of PCE ganglia trapped in glass-etched pore networks during the continuous injection of nZVI suspension or pure water. The nZVI injection in porous media is simulated as an advection- dispersion process by accounting for the attachment/detachment of nanoparticles on the pore-walls, and describing the kinetics of PCE dissolution and reaction by 1st order equations. Visualization experiments reveal that the gradual elimination of PCE ganglia by the injected nZVI is associated with the preferential "erosion" of the upstream interfacial regions. The step controlling the overall process kinetics might be either (i) the enhanced PCE dissolution or (ii) the direct reaction of bulk PCE with the nZVI deposited upon the ganglia interfaces. Inverse modeling of the experiments under the simplifying assumption of one active mechanism indicates that the estimated kinetic coefficients are increasing functions of the flow rate.

The accurate and sensitive determination of H2O2 is very important in many cases because it is a product of reactions catalysed by several oxidase enzymes in living cells and it is essential in environmental and pharmaceutical analyses. The fabrication of enzyme protein activity based biosensors is a very promising way for this purpose because the function of biological molecules is very specific, sensitive, and selective. Horseradish peroxidase (HRP) is the most commonly used enzyme for H2O2 detection because it can oxidize hydrogen atoms and, for example, xenobiotics in the presence of H2O2. In order to define the limit of detection (LOD) of H2O2 we made calibrations with guaiacol and amplex red (AR), which are hydrogen donors of HRP. The accumulation of the reaction products, tetraguaiacol, and resorufin, respectively, then can be easily detected by absorption or emission (fluorescence) spectroscopy. In our experiments an enzyme electrode was fabricated from ITO (indium tin oxide), functionalized multiwalled carbon nanotubes (f-MWCNTs), and HRP. Although the enzyme activity was smaller by about two orders of magnitude when the enzyme was bound to the f-MWCNTs (ca. 10−2 M H2O2/(M HRP·sec) compared to ca. 2 M H2O2/(M HRP·sec) and 5 M H2O2/(M HRP·sec) with AR and guaiacol in buffer solution), LOD of the H2O2 decomposition was about 6 pM H2O2/sec and 10 pM H2O2/sec in the case of AR and guaiacol, respectively.

The reaction center protein is the unit in the photosynthetic organisms in which the primary events of the photoelectric energy conversion take place during photosynthesis. When the photochemistry following the excitation of the reaction center by light is oversaturated there is a large probability to form reactive oxygen species (ROS, e.g., singlet oxygen (O-1(2))). Because the ROS components decrease the overall yield of the photochemical energy conversion there is a considerable effort in many laboratories to find conditions to reduce these harmful compounds. The aim of our work is to create a system by using carbon nanotubes (CNTs) and RCs for efficient light-energy conversion. The role of the O-1(2) that destroys the RC structure is discussed. 1,3-Diphenylisobenzofuran was used to detect the concentration of the O-1(2). Although, O-1(2) can be sensitized by CNTs in certain conditions, in our experiments the main sources of the O-1(2) are the RCs. The concentration of the O-1(2) in the equilibrium is determined by the forward sensitization and the backward deactivation processes of the components and functions of the CNT/RC composite. (C) 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

Since their discovery, carbon nanotubes (CNTs) have attracted intense attention to broad range of potential applications. In contrast to the 1D isolated single-walled carbon nanotubes (SWCNT), 2D films or bundles made of thousands of tubes have been introduced as more advantageous building blocks for new types of applications in mechanically flexible and stretchable, optically transparent electronic systems. In our experiments, we combined photosynthetic reaction centre proteins, the light energy converter units in living cells, purified from purple bacteria, with multiwalled carbon nanotube (MWCNT) bundles. The change in the conductivity of the bare MWCNT bundles and the RC/MWCNT composite after light excitation was measured and compared. We found that the electrical conductivity under light excitation depends on the intrinsic conductivity of individual tubes within the bundles and on structural characteristics, like geometry (diameter, length, spatial arrangement, interconnects, etc.) and the electronic coupling with the RCs. (C) 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

Photosensitizers may convert molecular oxygen into reactive oxygen species (ROS) including, e.g., singlet oxygen (O-1(2)), superoxide anion (O-2(-center dot)), and hydroxyl radicals ((OH)-O-center dot), chemicals with extremely high cyto- and potential genotoxicity. Photodynamic ROS reactions are determinative in medical photodynamic therapy (cancer treatment with externally added photosensitizers) and in reactions damaging the photosynthetic apparatus of plants (via internal pigments). The primary events of photosynthesis take place in the chlorophyll containing reaction center protein complex (RC), where the energy of light is converted into chemical potential. O-1(2) is formed by both bacterial bacteriochlorophylls and plant RC triplet chlorophylls in high light and if the quenching of O-1(2) is impaired. In plant physiology, reducing the formation of the ROS and thus lessening photooxidative membrane damage (including the RC protein) and increasing the efficiency of the photochemical energy conversion is of special interest. Carbon nanotubes, in artificial systems, are also known to react with singlet oxygen. To investigate the possibility of O-1(2) quenching by carbon nanotubes in a biological system, we studied the effect of carbon nanotubes on O-1(2) photogenerated by photosynthetic RCs purified from purple bacteria. 1,3-Diphenylisobenzofuran (DPBF), a dye responding to oxidation by O-1(2) with absorption decrease at 420nm was used to measure O-1(2) concentrations. O-1(2) was produced either from a photosensitizer (methylene blue) or from triplet photosynthetic RCs and the antioxidant capacity of carbon nanotubes was assessed. Less O-1(2) was detected by DPBF in the presence of carbon nanotubes, suggesting that these are potential quenchers of this ROS. (C) 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

H2O2 is a product of reactions catalysed by several oxidase enzymes and it is essential in environmental and pharmaceutical analyses. The most commonly used enzyme in understanding the biological behaviour of catalysed oxidation of H2O2 is horseradish peroxidase (HRP). In our experiments HRP was bound to carboxyl-functionalized multiwalled carbon nanotubes (MWNT-COOH) by N-hydroxysuccinimide (NHS) and 1-[3-dimethylaminopropyl]-3-ethyl-carbodiimide (EDC) crosslinkers. The activity of this bio-nanocomposite and the limit of detection (LOD) for H2O2 were determined by measuring the fluorescence of tetraguaiacol (which chemical is the product of guaiacol oxidation after addition of H2O2 to the reaction mixture) as a function of time. The hydrogen peroxide biosensor we developed exhibited a detection limit of 1.2 mu MH(2)O(2)s(-1) which resolution was better than the one measured in solution by about a factor of eight (it was 10 mu MH(2)O(2)s(-1) in solution). An attempt has been made to measure the concentration of H2O2 in an electrochemical cell with HRP immobilized on the surface of an electrode made of indium tin oxide (ITO, a transparent conductive oxide) and MWNT. (C) 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

Photosynthetic reaction center proteins purified from Rhodobacter sphaeroides purple bacterium were deposited on the surface of indium tin oxide (ITO), a transparent conductive oxide, and the photochemical/-physical properties of the composite were investigated. The kinetics of the light induced absorption change indicated that the RC was active in the composite and there was an interaction between the protein cofactors and the ITO. The electrochromic response of the bacteriopheophytine absorption at 771 nm showed an increased electric field perturbation around this chromophore on the surface of ITO compared to the one measured in solution. This absorption change is associated with the charge-compensating relaxation events inside the protein. Similar life time, but smaller magnitude of this absorption change was measured on the surface of borosilicate glass. The light induced change in the conductivity of the composite as a function of the concentration showed the typical sigmoid saturation characteristics unlike if the photochemically inactive chlorophyll was layered on the ITO. In this later case the light induced change in the conductivity was oppositely proportional to the chlorophyll concentration due to the thermal dissipation of the excitation energy. The sensitivity of the measurement is very high; few picomole RC can change the light induced resistance of the composite.

Photosynthetic reaction centres are membrane-spanning proteins, found in several classes of autotroph organisms, where a photoinduced charge separation and stabilization takes place with a quantum efficiency close to unity. The protein remains stable and fully functional also when extracted and purified in detergents thereby biotechnological applications are possible, for example, assembling it in nano-structures or in optoelectronic systems. Several types of bio-nanocomposite materials have been assembled by using reaction centres and different carrier matrices for different purposes in the field of light energy conversion (e. g., photovoltaics) or biosensing (e. g., for specific detection of pesticides). In this review we will summarize the current status of knowledge, the kinds of applications available and the difficulties to be overcome in the different applications. We will also show possible research directions for the close future in this specific field.

Specific inhibitory reactions of herbicides with photosynthetic reaction centers bound to working electrodes were monitored in a conventional electrochemical cell and a newly designed microfluidic electrochemical flow cell. In both cases, the bacterial reaction centers were bound to a transparent conductive metal oxide, indium-tin-oxide, electrode through carbon nanotubes. In the conventional cell, photocurrent densities of up to a few μA/cm2 could be measured routinely. The photocurrent could be blocked by the photosynthetic inhibitor terbutryn (I 50 = 0.38 ± 0.14 μM) and o-phenanthroline (I 50 = 63.9 ± 12.2 μM). The microfluidic flow cell device enabled us to reduce the sample volume and to simplify the electrode arrangement. The useful area of the electrodes remained the same (ca. 2 cm2), similar to the classical electrochemical cell; however, the size of the cell was reduced considerably. The microfluidic flow control enabled us monitoring in real time the binding/unbinding of the inhibitor and cofactor molecules at the secondary quinone site.

Basic principles of structural and functional requirements of photosynthetic energy conversion in hierarchically organized machineries are reviewed. Blueprints of photosynthesis, the energetic basis of virtually all life on Earth, can serve the basis for constructing artificial light energy-converting molecular devices. In photosynthetic organisms, the conversion of light energy into chemical energy takes places in highly organized fine-tunable systems with structural and functional hierarchy. The incident photons are absorbed by light-harvesting complexes, which funnel the excitation energy into reaction centre (RC) protein complexes containing redox-active chlorophyll molecules; the primary charge separations in the RCs are followed by vectorial transport of charges (electrons and protons) in the photosynthetic membrane. RCs possess properties that make their use in solar energy-converting and integrated optoelectronic systems feasible. Therefore, there is a large interest in many laboratories and in the industry toward their use in molecular devices. RCs have been bound to different carrier matrices, with their photophysical and photochemical activities largely retained in the nano-systems and with electronic connection to conducting surfaces. We show examples of RCs bound to carbon-based materials (functionalized and non-functionalized single- and multiwalled carbon nanotubes), transitional metal oxides (ITO) and conducting polymers and porous silicon and characterize their photochemical activities. Recently, we adapted several physical and chemical methods for binding RCs to different nanomaterials. It is generally found that the P(+)(QAQB)(-) charge pair, which is formed after single saturating light excitation is stabilized after the attachment of the RCs to the nanostructures, which is followed by slow reorganization of the protein structure. Measuring the electric conductivity in a direct contact mode or in electrochemical cell indicates that there is an electronic interaction between the protein and the inorganic carrier matrices. This can be a basis of sensing element of bio-hybrid device for biosensor and/or optoelectronic applications.

The long term stability and the redox interaction between single walled carbon nanotubes (SWNTs) and photosynthetic reaction center proteins (RCs) purified from purple bacterium Rhodobacter sphaeroides R-26 in the SWNT/RC complex has been investigated. The binding of SWNT to RC results in an accumulation of positive (the oxidized primary electron donor, P+) and negative (semiquinone forms, Q(A)(-) and Q(B)(-), the reduced primary and secondary quinones, respectively) charges followed by slow reorganization of the protein structure after excitation. The photochemical activity of the SWNT/RC complexes remains stable for several weeks even in dried form. In the absence of SWNT the secondary quinone activity decays quickly as a function of time after drying the RC onto a glass surface. Polarography measurements substantiate the idea that there is an electronic interaction between the RCs and SWNTs after light excitation, which was suggested earlier by optical measurements. The special electronic properties of the SWNT/protein complexes open the possibility for several applications, e. g., in microelectronics, analytics, or energy conversion and storage. (C) 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

Photosynthetic reaction center (RC) is one of the most important proteins, because it is Nature's solar battery converting light energy into chemical potential in the photosynthetic membrane assuring conditions for carbon reduction in cells. Although it is developed in nanometer scale, and is working in nanoscopic power, this is the protein that assures the energy input practically for the whole biosphere on Earth. The extremely large quantum yield of the primary charge separation (close to 100%) in the RC offers a big challenge to use it in nanodevices. Results of structural (AFM, EM), optical, and electro chemical investigations on RC bio-nanocomposite materials based on different carrier matrices (e.g., CNTs, ITO) will be presented.

The encapsulation of living plant cells into materials could offer the possibility to develop new green biochemical technologies. With the view to designing new functional materials, the physiological activity and cellular response of entrapped cells within different silica-based matrices have been assessed. A fine-tuning of the surface chemistry of the matrix has been achieved by the in situ copolymerization of an aqueous silica precursor and a biocompatible trifunctional silane bearing covalently bound neutral sugars. This method allows a facile control of chemical and physical interactions between the entrapped plant cells and the scaffold. The results show that the cell-matrix interaction has to be carefully controlled in order to avoid the mineralization of the cell wall which typically reduces the bioavailability of nutrients. Under appropriate conditions, the introduction of a trifunctional silane (ca. 10%) during the preparation of hybrid gels has shown to prolong the biological activity as well as the cellular viability of plant cells. The relations of cell behavior with some other key factors such as the porosity and the contraction of the matrix are also discussed.

International audience; Porous silicon microcavity (PSiMc) structures were used to immobilize photosynthetic reaction center (RC) purified from the purple bacterium Rhodobacter sphaeroides R-26. Two different binding methods were compared by specular reflectance measurements. Structural characterization of PSiMc was performed by Scanning Electron Microscopy (SEM) and Atomic Force Microscopy (AFM). The activity of the immobilized RC was checked by measuring the visible absorption spectra of the externally added electron donor, mammalian cytochrome c. PSi/RC complex was found to oxidize the cytochrome c after every saturating Xe flash, indicating the accessibility of specific surface binding sites on the immobilized RC, for the external electron donor. This new type of bio-nanomaterial is considered as an excellent model for new generation applications of silicon based electronics and biological redox systems.

International audience; The purified photosynthetic reaction center protein (RC) from Rhodobacter sphaeroides R-26 purple bacteria was bound to porous silicon microcavities (PSiMc) either through silane-glutaraldehyde (GTA) chemistry or via a noncovalent peptide cross-linker. The characteristic resonance mode in the microcavity reflectivity spectrum red shifted by several nanometers upon RC binding, indicating the protein infiltration into the porous silicon (PSi) photonic structure. Flash photolysis experiments confirmed the photochemical activity of RC after its binding to the solid substrate. The kinetic components of the intraprotein charge recombination were considerably faster (τfast = 14 (±9) ms, τslow = 230 (±28) ms with the RC bound through the GTA cross-linker and only τfast = 27 (±3) ms through peptide coating) than in solution (τfast = 120 (±3) ms, τslow = 1387 (±2) ms), indicating the effect of the PSi surface on the light-induced electron transfer in the protein. The PSi/RC complex was found to oxidize the externally added electron donor, mammalian cytochrome c, and the cytochrome oxidation was blocked by the competitive RC inhibitor, terbutryne. This fact indicates that the specific surface binding sites on the PSi-bound RC are still accessible to external cofactors and an electronic interaction with redox components in the aqueous environment is possible. This new type of biophotonic material is considered to be an excellent model for new generation applications at the interface of silicon-based electronics and biological redox systems designed by nature.