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Redox Enzymes (redox + enzyme)
Selected AbstractsRedox enzymes in the plant plasma membrane and their possible rolesPLANT CELL & ENVIRONMENT, Issue 12 2000A. Bérczi ABSTRACT Purified plasma membrane (PM) vesicles from higher plants contain redox proteins with low-molecular-mass prosthetic groups such as flavins (both FMN and FAD), hemes, metals (Cu, Fe and Mn), thiol groups and possibly naphthoquinone (vitamin K1), all of which are likely to participate in redox processes. A few enzymes have already been identified: Monodehydroascorbate reductase (EC 1.6.5.4) is firmly bound to the cytosolic surface of the PM where it might be involved in keeping both cytosolic and, together with a b -type cytochrome, apoplastic ascorbate reduced. A malate dehydrogenase (EC 1.1.1.37) is localized on the inner side of the PM. Several NAD(P)H-quinone oxidoreductases have been purified from the cytocolic surface of the PM, but their function is still unknown. Different forms of nitrate reductase (EC 1.6.6.1,3) are found attached to, as well as anchored in, the PM where they may act as a nitrate sensor and/or contribute to blue-light perception, although both functions are speculative. Ferric-chelate-reducing enzymes (EC 1.6.99.13) are localized and partially characterized on the inner surface of the PM but they may participate only in the reduction of ferric-chelates in the cytosol. Very recently a ferric-chelate-reducing enzyme containing binding sites for FAD, NADPH and hemes has been identified and suggested to be a trans -PM protein. This enzyme is involved in the reduction of apoplastic iron prior to uptake of Fe2+ and is induced by iron deficiency. The presence of an NADPH oxidase, similar to the so-called respiratory burst oxidase in mammals, is still an open question. An auxin-stimulated and cyanide-insensitive NADH oxidase (possibly a protein disulphide reductase) has been characterized but its identity is still awaiting independent confirmation. Finally, the only trans -PM redox protein which has been partially purified from plant PM so far is a high-potential and ascorbate-reducible b -type cytochrome. In co-operation with vitamin K1 and an NAD(P)H-quinone oxidoreductase, it may participate in trans -PM electron transport. [source] Gold Nanoparticle-Based Mediatorless Biosensor Prepared on Microporous ElectrodeELECTROANALYSIS, Issue 3 2006Fenghua Zhang Abstract A mediatorless biosensor was fabricated with a double-sided microporous gold electrode by successively immobilizing a mixed self-assembled monolayer (SAM) comprising carboxylic-acid- and thiol-terminated thiolate (dl -thiorphan and 1,8-octanedithiol), glucose oxidase (GOx) and finally gold nanoparticle (Au NP) on one working side. The double-sided microporous gold electrodes were formed by plasma sputtering of gold on a porous nylon substrate, yielding a face-to-face type two-electrode electrochemical cell. While the straight chain molecule 1,8-octanedithiol forms a dense insulating monolayer, the side armed dl -thiorphan forms a low density layer for the diffusion of redox couples to the electrode surface. The mixed SAM not only provided the linking functional groups for both enzyme and Au NP but also resulted in the appropriately spaced monolayer for direct electron tansfer (ET) process from the center of the redox enzyme to the electrode surface. After covalently immobilizing GOx onto the carboxylic-acid-terminated monolayer, Au NP was easily immobilized to both enzyme and nearby thiols by simple dispensing of the colloidal gold solution. It was observed that the resulting amperometric biosensor exhibited quantitatively the same response to glucose in the presence and in the absence of dissolved oxygen, which evidence that the Au NPs immobilized on and around the GOx promote direct ET from the enzymes to the electrode, assuming the role of a common redox mediator. [source] Light-driven Hydrogen Production by a Hybrid Complex of a [NiFe]-Hydrogenase and the Cyanobacterial Photosystem IPHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 3 2006Masaki Ihara ABSTRACT In order to generate renewable and clean fuels, increasing efforts are focused on the exploitation of photosynthetic microorganisms for the production of molecular hydrogen from water and light. In this study we engineered a ,hard-wired' protein complex consisting of a hydrogenase and photosystem I (hydrogenase-PSI complex) as a direct light-to-hydrogen conversion system. The key component was an artificial fusion protein composed of the membrane-bound [NiFe] hydrogenase from the ,-proteobacterium Ralstonia eutropha H16 and the peripheral PSI subunit PsaE of the cyanobacterium Thermosy-nechococcus elongatus. The resulting hydrogenase-PsaE fusion protein associated with PsaE-free PSI spontaneously, thereby forming a hydrogenase-PSI complex as confirmed by sucrosegradient ultracentrifuge and immunoblot analysis. The hydrogenase-PSI complex displayed light-driven hydrogen production at a rate of 0.58 ,mol H2ˇ mg chlorophyll,1ˇ h,1. The complex maintained its accessibility to the native electron acceptor ferredoxin. This study provides the first example of a light-driven enzymatic reaction by an artificial complex between a redox enzyme and photosystem I and represents an important step on the way to design a photosynthetic organism that efficiently converts solar energy and water into hydrogen. [source] Wiring Enzymes in Nanostructures Built with Electrostatically Self-Assembled Thin FilmsCHEMPHYSCHEM, Issue 1 2005Ernesto J. Calvo Prof. Abstract The construction of electrostatically self-assembled intelligent nanostructures on electrodes with redox enzyme layers and redox polymer molecular wires defined in space allowed the analysis of redox charge transport from the redox enzyme to the electrode along nanometric distances. Recent results on the electrical connection of enzymes to electrodes and perspectives of generating electrical signals from molecular recognition in integrated enzyme electrodes are discussed. [source] Functional properties of the protein disulfide oxidoreductase from the archaeon Pyrococcus furiosusFEBS JOURNAL, Issue 16 2004A member of a novel protein family related to protein disulfide-isomerase Protein disulfide oxidoreductases are ubiquitous redox enzymes that catalyse dithiol,disulfide exchange reactions with a CXXC sequence motif at their active site. A disulfide oxidoreductase, a highly thermostable protein, was isolated from Pyrococcus furiosus (PfPDO), which is characterized by two redox sites (CXXC) and an unusual molecular mass. Its 3D structure at high resolution suggests that it may be related to the multidomain protein disulfide-isomerase (PDI), which is currently known only in eukaryotes. This work focuses on the functional characterization of PfPDO as well as its relation to the eukaryotic PDIs. Assays of oxidative, reductive, and isomerase activities of PfPDO were performed, which revealed that the archaeal protein not only has oxidative and reductive activity, but also isomerase activity. On the basis of structural data, two single mutants (C35S and C146S) and a double mutant (C35S/C146S) of PfPDO were constructed and analyzed to elucidate the specific roles of the two redox sites. The results indicate that the CPYC site in the C-terminal half of the protein is fundamental to reductive/oxidative activity, whereas isomerase activity requires both active sites. In comparison with PDI, the ATPase activity was tested for PfPDO, which was found to be cation-dependent with a basic pH optimum and an optimum temperature of 90 °C. These results and an investigation on genomic sequence databases indicate that PfPDO may be an ancestor of the eukaryotic PDI and belongs to a novel protein disulfide oxidoreductase family. [source] Productive Asymmetric Styrene Epoxidation Based on a Next Generation Electroenzymatic MethodologyADVANCED SYNTHESIS & CATALYSIS (PREVIOUSLY: JOURNAL FUER PRAKTISCHE CHEMIE), Issue 14-15 2009Reto Ruinatscha Abstract We have established a novel and scalable methodology for the productive coupling of redox enzymes to reductive electrochemical cofactor regeneration relying on efficient mass transfer of the cofactor to the electron-delivering cathode. Proof of concept is provided by styrene monooxygenase (StyA) catalyzing the asymmetric (S)-epoxidation of styrene with high enantiomeric excess, space-time yields, and current efficiencies. Highly porous reticulated vitreous carbon electrodes, maximized in volumetric surface area, were employed in a flow-through mode to rapidly regenerate the consumed FADH2 cofactor required for StyA activity. A systematic investigation of the parameters determining cofactor mass transfer revealed that low FAD concentrations and high flow rates enabled the continuous synthesis of the product (S)-styrene oxide at high rates, while at the same time the accumulation of the side-products acetophenone and phenylacetaldehyde was minimized. At 10,,M FAD and a flow rate of 150,mL,min,1, an average space-time yield of 0.35,g,L,1,h,1 could be achieved during 2,h with a final (S)-styrene oxide yield of 75.2%. At two-fold lower aeration rates, the electroenzymatic reaction could be sustained for 12,h, albeit at the expense of lower (59%) overall space-time yields. Under these conditions, as much as 20.5% of the utilized current could be channeled into (S)-styrene oxide formation. In comparison with state-of-the-art electroenzymatic methodologies for the same conversion, (S)-styrene oxide synthesis could be improved up to 150-fold with respect to both reaction time and space-time yield. These productivities constitute the most efficient reaction reported for asymmetric in vitro epoxidations of styrene. [source] Rapid breakdown of exogenous extracellular hydrogen peroxide by lichensPHYSIOLOGIA PLANTARUM, Issue 3 2007Richard P. Beckett All organisms, even highly stress-tolerant lichens, produce a variety of reactive oxygen species (ROS) during and after stress. Furthermore, the cell walls of some lichens in Suborder Peltigerineae contain laccases, and therefore can produce quinone radicals that can break down to yield ROS. While the extracellular ROS produced by these enzymes probably play important roles in the biology of these lichens, they may also be potentially harmful and need to be rapidly broken down. To test this, rates of breakdown of exogenously supplied H2O2 were measured in a range of lichen species. Considerable diversity existed in rates of H2O2 breakdown but rates were on average almost double in members of Suborder Peltigerineae. While all lichens tested appeared to lack extracellular peroxidases and catalases, enzymes normally involved in breaking down H2O2, extracellular tyrosinase activity could be readily detected in the Peltigerineae. A role for tyrosinases in H2O2 breakdown was supported by the results from experiments involving inhibitors, and demonstration of the simultaneous release into an incubation solution of tyrosinase activity and the ability to breakdown H2O2. Rates of breakdown were very high, and tyrosinase appeared to break down H2O2 by a catalase-like mechanism. However, significant rates of breakdown of H2O2 also occurred in species that did not possess cell wall redox enzymes. These species probably took up the exogenously supplied H2O2 intracellularly and then broke it down by the usual catalases and peroxidases. The importance of H2O2 degradation is discussed in terms of its possible role in defence against the harmful effects of ROS. [source] CxxS: Fold-independent redox motif revealed by genome-wide searches for thiol/disulfide oxidoreductase functionPROTEIN SCIENCE, Issue 10 2002Dmitri E. Fomenko Abstract Redox reactions involving thiol groups in proteins are major participants in cellular redox regulation and antioxidant defense. Although mechanistically similar, thiol-dependent redox processes are catalyzed by structurally distinct families of enzymes, which are difficult to identify by available protein function prediction programs. Herein, we identified a functional motif, CxxS (cysteine separated from serine by two other residues), that was often conserved in redox enzymes, but rarely in other proteins. Analyses of complete Escherichia coli, Campylobacter jejuni, Methanococcus jannaschii, and Saccharomyces cerevisiae genomes revealed a high proportion of proteins known to use the CxxS motif for redox function. This allowed us to make predictions in regard to redox function and identity of redox groups for several proteins whose function previously was not known. Many proteins containing the CxxS motif had a thioredoxin fold, but other structural folds were also present, and CxxS was often located in these proteins upstream of an ,-helix. Thus, a conserved CxxS sequence followed by an ,-helix is typically indicative of a redox function and corresponds to thiol-dependent redox sites in proteins. The data also indicate a general approach of genome-wide identification of redox proteins by searching for simple conserved motifs within secondary structure patterns. [source] Redox proteins in mammalian cell death: an evolutionarily conserved function in mitochondria and prokaryotesCELLULAR MICROBIOLOGY, Issue 4 2003Vasu Punj Summary Mammalian cell mitochondria are believed to have prokaryotic ancestry. Mitochondria are not only the powerhouse of energy generation within the eukaryotic cell but they also play a major role in inducing apoptotic cell death through release of redox proteins such as cytochrome c and the apoptosis-inducing factor (AIF), a flavoprotein with NADH oxidase activity. Recent evidence indicates that some present day prokaryotes release redox proteins that induce apoptosis in mammalian cells through stabilization of the tumour suppressor protein p53. p53 interacts with mitochondria either directly or through activation of the genes for pro-apoptotic proteins such as Bax or NOXA or genes that encode redox enzymes responsible for the production of reactive oxygen species (ROS). The analogy between the ancient ancestors of present day bacteria, the mitochondria, and the present day bacteria with regard to their ability to release redox proteins for triggering mammalian cell death is an interesting example of functional conservation during the hundreds of millions of years of evolution. It is possible that the ancestors of the present day prokaryotes released redox proteins to kill the ancestors of the eukaryotes. During evolution of the mitochondria from prokaryotes as obligate endosymbionts, the mitochondria maintained the same functions to programme their own host cell death. [source] | |||||||||||||