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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

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.

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.

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 proteins (RCs) are the most efficient light energy converter systems in nature. The first steps of the primary charge separation in photosynthesis take place in these proteins. Due to their unique properties, combining RCs with nano-structures promising applications can be predicted in optoelectronic systems. In the present work RCs purified from Rhodobacter sphaeroides purple bacteria were immobilized on multiwalled carbon nanotubes (CNTs). Carboxyl—and amine-functionalised CNTs were used, so different binding procedures, physical sorption and chemical sorption as well, could be applied as immobilization techniques. Light-induced singlet oxygen production was measured in the prepared photoactive biocomposites in water-based suspension by histidine mediated chemical trapping. Carbon nanotubes were applied under different conditions in order to understand their role in the equilibration of singlet oxygen concentration in the suspension. CNTs acted as effective quenchers of 1O2 either by physical (resonance) energy transfer or by chemical (oxidation) reaction and their efficiency showed dependence on the diffusion distance of 1O2.

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.