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Russ Hille - One of the best experts on this subject based on the ideXlab platform.
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crystallographic and kinetic analyses of the fdsbg subcomplex of the cytosolic Formate Dehydrogenase fdsabg from cupriavidus necator
2020Co-Authors: Tynan Young, Russ Hille, Dimitri Niks, Sheron Hakopian, Timothy K Tam, Gregor BlahaAbstract:Formate oxidation to carbon dioxide is a key reaction in one-carbon compound metabolism, and its reverse reaction represents the first step in carbon assimilation in the acetogenic and methanogenic branches of many anaerobic organisms. The molybdenum-containing Dehydrogenase FdsABG is a soluble NAD+-dependent Formate Dehydrogenase and a member of the NADH Dehydrogenase superfamily. Here, we present the first structure of the FdsBG subcomplex of the cytosolic FdsABG Formate Dehydrogenase from the hydrogen-oxidizing bacterium Cupriavidus necator H16 both with and without bound NADH. The structures revealed that the two iron-sulfur clusters, Fe4S4 in FdsB and Fe2S2 in FdsG, are closer to the FMN than they are in other NADH Dehydrogenases. Rapid kinetic studies and EPR measurements of rapid freeze-quenched samples of the NADH reduction of FdsBG identified a neutral flavin semiquinone, FMNH•, not previously observed to participate in NADH-mediated reduction of the FdsABG holoenzyme. We found that this semiquinone forms through the transfer of one electron from the fully reduced FMNH−, initially formed via NADH-mediated reduction, to the Fe2S2 cluster. This Fe2S2 cluster is not part of the on-path chain of iron-sulfur clusters connecting the FMN of FdsB with the active-site molybdenum center of FdsA. According to the NADH-bound structure, the nicotinamide ring stacks onto the re-face of the FMN. However, NADH binding significantly reduced the electron density for the isoalloxazine ring of FMN and induced a conformational change in residues of the FMN-binding pocket that display peptide-bond flipping upon NAD+ binding in proper NADH Dehydrogenases.
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crystallographic and kinetic analyses of the fdsbg subcomplex of the cytosolic Formate Dehydrogenase fdsabg from cupriavidus necator
2020Co-Authors: Tynan Young, Russ Hille, Dimitri Niks, Xuejun Yu, Sheron Hakopian, Gregor BlahaAbstract:: Formate oxidation to carbon dioxide is a key reaction in one-carbon compound metabolism, and its reverse reaction represents the first step in carbon assimilation in the acetogenic and methanogenic branches of many anaerobic organisms. The molybdenum-containing Dehydrogenase FdsABG is a soluble NAD+-dependent Formate Dehydrogenase and a member of the NADH Dehydrogenase superfamily. Here, we present the first structure of the FdsBG subcomplex of the cytosolic FdsABG Formate Dehydrogenase from the hydrogen-oxidizing bacterium Cupriavidus necator H16 both with and without bound NADH. The structures revealed that the two iron-sulfur clusters, Fe4S4 in FdsB and Fe2S2 in FdsG, are closer to the FMN than they are in other NADH Dehydrogenases. Rapid kinetic studies and EPR measurements of rapid freeze-quenched samples of the NADH reduction of FdsBG identified a neutral flavin semiquinone, FMNH•, not previously been observed to participate in NADH-mediated reduction of the FdsABG holoenzyme. We found that this semiquinone forms through the transfer of one electron from the fully reduced FMNH , initially formed via NADH-mediated reduction, to the Fe2S2 cluster. This Fe2S2 cluster is not part of the on-path chain of iron-sulfur clusters connecting the FMN of FdsB with the active-site molybdenum-center of FdsA. According to the NADH-bound structure, the nicotinamide ring stacks onto the re-face of the FMN. However, NADH binding significantly reduced the electron density for the isoalloxazine ring of FMN and induced a conformational change in residues of the FMN-binding pocket that display peptide-bond flipping upon NAD+ binding in proper NADH Dehydrogenases.
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deconvolution of reduction potentials of Formate Dehydrogenase from cupriavidus necator
2019Co-Authors: Lindsey M Walker, Russ Hille, Dimitri Niks, Bin Li, Sean J ElliottAbstract:: The Formate Dehydrogenase enzyme from Cupriavidus necator (FdsABG) carries out the two-electron oxidation of Formate to CO2, but is also capable of reducing CO2 back to Formate, a potential biofuel. FdsABG is a heterotrimeric enzyme that performs this transformation using nine redox-active cofactors: a bis(molybdopterin guanine dinucleotide) (bis-MGD) at the active site coupled to seven iron-sulfur clusters, and one equivalent of flavin mononucleotide (FMN). To better understand the pathway of electron flow in FdsABG, the reduction potentials of the various cofactors were examined through direct electrochemistry. Given the redundancy of cofactors, a truncated form of the FdsA subunit was developed that possesses only the bis-MGD active site and a singular [4Fe-4S] cluster. Electrochemical characterization of FdsABG compared to truncated FdsA shows that the measured reduction potentials are remarkably similar despite the truncation with two observable features at - 265 mV and - 455 mV vs SHE, indicating that the voltammetry of the truncated enzyme is representative of the reduction potentials of the intact heterotrimer. By producing truncated FdsA without the necessary maturation factors required for bis-MGD insertion, a form of the truncated FdsA that possesses only the [4Fe-4S] was produced, which gives a single voltammetric feature at - 525 mV, allowing the contributions of the molybdenum cofactor to be associated with the observed feature at - 265 mV. This method allowed for the deconvolution of reduction potentials for an enzyme with highly complex cofactor content to know more about the thermodynamic landscape of catalysis.
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efficient reduction of co2 by the molybdenum containing Formate Dehydrogenase from cupriavidus necator ralstonia eutropha
2017Co-Authors: Xuejun Yu, Dimitri Niks, Ashok Mulchandani, Russ HilleAbstract:Abstract The ability of the FdsABG Formate Dehydrogenase from Cupriavidus necator (formerly known as Ralstonia eutropha) to catalyze the reverse of the physiological reaction, the reduction of CO2 to Formate utilizing NADH as electron donor, has been investigated. Contrary to previous studies of this enzyme, we demonstrate that it is in fact effective in catalyzing the reverse reaction, with a kcat of 11 ± 0.4 s-1. We also quantify the stoichiometric accumulation of formic acid as the product of the reaction and demonstrate that the observed kinetic parameters for catalysis in the forward and reverse reaction are thermodynamically consistent, complying with the expected Haldane relationships. Finally, we demonstrate the reaction conditions necessary for gauging the ability of a given Formate Dehydrogenase or other CO2-utilizing enzyme to catalyze the reverse direction so as to avoid false negative results. In conjunction with our earlier studies on the reaction mechanism of this enzyme (Niks et al. (2016) J. Biol. Chem. 291, 1162- 1174), and on the basis of the present work we conclude that all molybdenum- and tungsten-containing Formate Dehydrogenases and related enzymes likely operate via a simple hydride transfer mechanism and are effective in catalysing the reversible interconversion of CO2 and Formate under the appropriate experimental conditions.
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efficient reduction of co2 by the molybdenum containing Formate Dehydrogenase from cupriavidus necator ralstonia eutropha
2017Co-Authors: Dimitri Niks, Ashok Mulchandani, Russ HilleAbstract:The ability of the FdsABG Formate Dehydrogenase from Cupriavidus necator (formerly known as Ralstonia eutropha) to catalyze the reverse of the physiological reaction, the reduction of CO2 to Formate utilizing NADH as electron donor, has been investigated. Contrary to previous studies of this enzyme, we demonstrate that it is in fact effective in catalyzing the reverse reaction with a kcat of 11 ± 0.4 s−1. We also quantify the stoichiometric accumulation of formic acid as the product of the reaction and demonstrate that the observed kinetic parameters for catalysis in the forward and reverse reactions are thermodynamically consistent, complying with the expected Haldane relationships. Finally, we demonstrate the reaction conditions necessary for gauging the ability of a given Formate Dehydrogenase or other CO2-utilizing enzyme to catalyze the reverse direction to avoid false negative results. In conjunction with our earlier studies on the reaction mechanism of this enzyme and on the basis of the present work, we conclude that all molybdenum- and tungsten-containing Formate Dehydrogenases and related enzymes likely operate via a simple hydride transfer mechanism and are effective in catalyzing the reversible interconversion of CO2 and Formate under the appropriate experimental conditions.
Dimitri Niks - One of the best experts on this subject based on the ideXlab platform.
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crystallographic and kinetic analyses of the fdsbg subcomplex of the cytosolic Formate Dehydrogenase fdsabg from cupriavidus necator
2020Co-Authors: Tynan Young, Russ Hille, Dimitri Niks, Sheron Hakopian, Timothy K Tam, Gregor BlahaAbstract:Formate oxidation to carbon dioxide is a key reaction in one-carbon compound metabolism, and its reverse reaction represents the first step in carbon assimilation in the acetogenic and methanogenic branches of many anaerobic organisms. The molybdenum-containing Dehydrogenase FdsABG is a soluble NAD+-dependent Formate Dehydrogenase and a member of the NADH Dehydrogenase superfamily. Here, we present the first structure of the FdsBG subcomplex of the cytosolic FdsABG Formate Dehydrogenase from the hydrogen-oxidizing bacterium Cupriavidus necator H16 both with and without bound NADH. The structures revealed that the two iron-sulfur clusters, Fe4S4 in FdsB and Fe2S2 in FdsG, are closer to the FMN than they are in other NADH Dehydrogenases. Rapid kinetic studies and EPR measurements of rapid freeze-quenched samples of the NADH reduction of FdsBG identified a neutral flavin semiquinone, FMNH•, not previously observed to participate in NADH-mediated reduction of the FdsABG holoenzyme. We found that this semiquinone forms through the transfer of one electron from the fully reduced FMNH−, initially formed via NADH-mediated reduction, to the Fe2S2 cluster. This Fe2S2 cluster is not part of the on-path chain of iron-sulfur clusters connecting the FMN of FdsB with the active-site molybdenum center of FdsA. According to the NADH-bound structure, the nicotinamide ring stacks onto the re-face of the FMN. However, NADH binding significantly reduced the electron density for the isoalloxazine ring of FMN and induced a conformational change in residues of the FMN-binding pocket that display peptide-bond flipping upon NAD+ binding in proper NADH Dehydrogenases.
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crystallographic and kinetic analyses of the fdsbg subcomplex of the cytosolic Formate Dehydrogenase fdsabg from cupriavidus necator
2020Co-Authors: Tynan Young, Russ Hille, Dimitri Niks, Xuejun Yu, Sheron Hakopian, Gregor BlahaAbstract:: Formate oxidation to carbon dioxide is a key reaction in one-carbon compound metabolism, and its reverse reaction represents the first step in carbon assimilation in the acetogenic and methanogenic branches of many anaerobic organisms. The molybdenum-containing Dehydrogenase FdsABG is a soluble NAD+-dependent Formate Dehydrogenase and a member of the NADH Dehydrogenase superfamily. Here, we present the first structure of the FdsBG subcomplex of the cytosolic FdsABG Formate Dehydrogenase from the hydrogen-oxidizing bacterium Cupriavidus necator H16 both with and without bound NADH. The structures revealed that the two iron-sulfur clusters, Fe4S4 in FdsB and Fe2S2 in FdsG, are closer to the FMN than they are in other NADH Dehydrogenases. Rapid kinetic studies and EPR measurements of rapid freeze-quenched samples of the NADH reduction of FdsBG identified a neutral flavin semiquinone, FMNH•, not previously been observed to participate in NADH-mediated reduction of the FdsABG holoenzyme. We found that this semiquinone forms through the transfer of one electron from the fully reduced FMNH , initially formed via NADH-mediated reduction, to the Fe2S2 cluster. This Fe2S2 cluster is not part of the on-path chain of iron-sulfur clusters connecting the FMN of FdsB with the active-site molybdenum-center of FdsA. According to the NADH-bound structure, the nicotinamide ring stacks onto the re-face of the FMN. However, NADH binding significantly reduced the electron density for the isoalloxazine ring of FMN and induced a conformational change in residues of the FMN-binding pocket that display peptide-bond flipping upon NAD+ binding in proper NADH Dehydrogenases.
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deconvolution of reduction potentials of Formate Dehydrogenase from cupriavidus necator
2019Co-Authors: Lindsey M Walker, Russ Hille, Dimitri Niks, Bin Li, Sean J ElliottAbstract:: The Formate Dehydrogenase enzyme from Cupriavidus necator (FdsABG) carries out the two-electron oxidation of Formate to CO2, but is also capable of reducing CO2 back to Formate, a potential biofuel. FdsABG is a heterotrimeric enzyme that performs this transformation using nine redox-active cofactors: a bis(molybdopterin guanine dinucleotide) (bis-MGD) at the active site coupled to seven iron-sulfur clusters, and one equivalent of flavin mononucleotide (FMN). To better understand the pathway of electron flow in FdsABG, the reduction potentials of the various cofactors were examined through direct electrochemistry. Given the redundancy of cofactors, a truncated form of the FdsA subunit was developed that possesses only the bis-MGD active site and a singular [4Fe-4S] cluster. Electrochemical characterization of FdsABG compared to truncated FdsA shows that the measured reduction potentials are remarkably similar despite the truncation with two observable features at - 265 mV and - 455 mV vs SHE, indicating that the voltammetry of the truncated enzyme is representative of the reduction potentials of the intact heterotrimer. By producing truncated FdsA without the necessary maturation factors required for bis-MGD insertion, a form of the truncated FdsA that possesses only the [4Fe-4S] was produced, which gives a single voltammetric feature at - 525 mV, allowing the contributions of the molybdenum cofactor to be associated with the observed feature at - 265 mV. This method allowed for the deconvolution of reduction potentials for an enzyme with highly complex cofactor content to know more about the thermodynamic landscape of catalysis.
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efficient reduction of co2 by the molybdenum containing Formate Dehydrogenase from cupriavidus necator ralstonia eutropha
2017Co-Authors: Xuejun Yu, Dimitri Niks, Ashok Mulchandani, Russ HilleAbstract:Abstract The ability of the FdsABG Formate Dehydrogenase from Cupriavidus necator (formerly known as Ralstonia eutropha) to catalyze the reverse of the physiological reaction, the reduction of CO2 to Formate utilizing NADH as electron donor, has been investigated. Contrary to previous studies of this enzyme, we demonstrate that it is in fact effective in catalyzing the reverse reaction, with a kcat of 11 ± 0.4 s-1. We also quantify the stoichiometric accumulation of formic acid as the product of the reaction and demonstrate that the observed kinetic parameters for catalysis in the forward and reverse reaction are thermodynamically consistent, complying with the expected Haldane relationships. Finally, we demonstrate the reaction conditions necessary for gauging the ability of a given Formate Dehydrogenase or other CO2-utilizing enzyme to catalyze the reverse direction so as to avoid false negative results. In conjunction with our earlier studies on the reaction mechanism of this enzyme (Niks et al. (2016) J. Biol. Chem. 291, 1162- 1174), and on the basis of the present work we conclude that all molybdenum- and tungsten-containing Formate Dehydrogenases and related enzymes likely operate via a simple hydride transfer mechanism and are effective in catalysing the reversible interconversion of CO2 and Formate under the appropriate experimental conditions.
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efficient reduction of co2 by the molybdenum containing Formate Dehydrogenase from cupriavidus necator ralstonia eutropha
2017Co-Authors: Dimitri Niks, Ashok Mulchandani, Russ HilleAbstract:The ability of the FdsABG Formate Dehydrogenase from Cupriavidus necator (formerly known as Ralstonia eutropha) to catalyze the reverse of the physiological reaction, the reduction of CO2 to Formate utilizing NADH as electron donor, has been investigated. Contrary to previous studies of this enzyme, we demonstrate that it is in fact effective in catalyzing the reverse reaction with a kcat of 11 ± 0.4 s−1. We also quantify the stoichiometric accumulation of formic acid as the product of the reaction and demonstrate that the observed kinetic parameters for catalysis in the forward and reverse reactions are thermodynamically consistent, complying with the expected Haldane relationships. Finally, we demonstrate the reaction conditions necessary for gauging the ability of a given Formate Dehydrogenase or other CO2-utilizing enzyme to catalyze the reverse direction to avoid false negative results. In conjunction with our earlier studies on the reaction mechanism of this enzyme and on the basis of the present work, we conclude that all molybdenum- and tungsten-containing Formate Dehydrogenases and related enzymes likely operate via a simple hydride transfer mechanism and are effective in catalyzing the reversible interconversion of CO2 and Formate under the appropriate experimental conditions.
V I Tishkov - One of the best experts on this subject based on the ideXlab platform.
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advantages of Formate Dehydrogenase reaction for efficient nad quantification in biological samples
2020Co-Authors: Artem V Artiukhov, V I Tishkov, A A Pometun, Sofia A Zubanova, Victoria I BunikAbstract:The medical significance of NAD+-dependent metabolic regulation acquires increasing attention, demanding rapid and clinically feasible quantification of NAD+ in complex biological samples. Here we describe the usage of Formate Dehydrogenase for a straightforward and highly specific fluorometric assay of NAD+ in tissue extracts, not requiring chromatographic separation of nucleotides. The assay employs the irreversible reaction of Formate oxidation coupled to NAD+ reduction, catalyzed by the enzyme which has high affinity and specificity to NAD+, and is stable under a variety of conditions. The assay reliably quantifies NAD+ in the methanol extracts of the rat brain cortex and mitochondria.
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Influence of His_6 Sequence on the Properties of Formate Dehydrogenase from Bacterium Pseudomonas sp. 101
2020Co-Authors: A A Pometun, I. V. Uporov, S S Savin, P. D. Parshin, N. P. Galanicheva, D. L. Atroshenko, V I TishkovAbstract:NAD(P)^+-dependent Formate Dehydrogenase (FDH, EC 1.2.1.2.) is actively used in processes of chiral synthesis by oxidoreductases with systems of reduced cofactor regeneration. The efficient use of FDH in such systems requires simple and fast enzyme purification. Metal-chelate affinity chromatography is widely used for such purposes. The method requires the presence of at least six His residues at N- or C-terminus of protein. The addition of extra His residues can affect enzyme properties. The computer modeling of the structure of FDH from bacterium Pseudomonas sp. 101 with different positions of His_6 sequence showed that the optimal case is His-tag at N-terminus. Three types of PseFDH with His_6 were prepared: wild-type NAD^+-dependent enzyme and two mutant NADP^+-specific forms. New PseFDHs were obtained as homogeneous preparations through a one-step purification procedure. The comparison of PseFDHs with and without His-tag showed that they have similar kinetic properties.
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improvement of the soy Formate Dehydrogenase properties by rational design
2015Co-Authors: I S Kargov, V I Tishkov, A A Alekseeva, S S Savin, S Y U KleimenovAbstract:Previous experiments on substitution of the residue Phe290 to Asp, Asn and Ser in NAD(+)-dependent Formate Dehydrogenase from soya Glycine max (SoyFDH) showed important role of the residue in enzyme thermal stability and catalytic properties (Alekseeva et al. Prot. Eng. Des. Sel., 2012a; 25: :781-88). In this work, we continued site-directed mutagenesis experiments of the Phe290 and the residue was changed to Ala, Thr, Tyr, Glu and Gln. All amino acid changes resulted in increase of catalytic constant from 2.9 to 3.5-4.7 s(-1). The substitution Phe290Ala led to KM (NAD+) decrease from 13.3 to 8.6 μM, and substitutions Phe290Tyr and Phe290Glu resulted in decrease and increase of KM (HCOO-) from 1.5 to 0.9 and -2.9 mM, respectively. The highest improvement of catalytic properties was observed for SoyFDH Phe290Ala which showed 2-fold higher catalytic efficiency with both substrates. Stability of mutants was examined by study of thermal inactivation kinetics and differential scanning calorimetry (DSC). All five amino acids provided increase of thermal stability of mutant SoyFDH in comparison with wild-type enzyme. Mutant SoyFDH Phe290Glu showed the highest improvement-the stabilization effect was 43 at 56°C. The DSC data agree with results of thermal inactivation kinetics. Substitutions Phe290Tyr, Phe290Thr, Phe290Gln and Phe290Glu provided Tm value increase 0.6°-6.6°. SoyFDH Phe290Glu and previously prepared SoyFDH Phe290Asp show similar thermal stability as enzymes from Candida boidinii and Mycobacterium vaccae N10 and have higher catalytic efficiency with NAD(+) compared with all described FDHs. Therefore, these mutants are very perspective enzymes for coenzyme regeneration in processes of chiral synthesis with Dehydrogenases.
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nad dependent Formate Dehydrogenase from plants
2011Co-Authors: A A Alekseeva, S S Savin, V I TishkovAbstract:NAD + -dependent Formate Dehydrogenase (FDH, EC 1.2.1.2) widely occurs in nature. FDH consists of two identical subunits and contains neither prosthetic groups nor metal ions. This type of FDH was found in different microorganisms (including pathogenic ones), such as bacteria, yeasts, fungi, and plants. As opposed to microbiological FDHs functioning in cytoplasm, plant FDHs localize in mitochondria. Formate Dehydrogenase activity was first discovered as early as in 1921 in plant; however, until the past decade FDHs from plants had been considerably less studied than the enzymes from microorganisms. This review summarizes the recent results on studying the physiological role, properties, structure, and protein engineering of plant Formate dehy- drogenases.
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protein engineering of Formate Dehydrogenase
2006Co-Authors: V I Tishkov, V PopovAbstract:NAD + -dependent Formate Dehydrogenase (FDH, EC 1.2.1.2) is one of the best enzymes for the purpose of NADH regeneration in Dehydrogenase-based synthesis of optically active compounds. Low operational stability and high production cost of native FDHs limit their applicationincommercial productionofchiralcompounds.ThereviewsummarizestheresultsonengineeringofbacterialandyeastFDHsaimedat improving their chemical and thermal stability, catalytic activity, switch in coenzyme specificity from NAD + to NADP + and overexpression in
Yutaka Amao - One of the best experts on this subject based on the ideXlab platform.
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can Formate Dehydrogenase from candida boidinii catalytically reduce carbon dioxide bicarbonate or carbonate to Formate
2020Co-Authors: Ryohei Sato, Yutaka AmaoAbstract:Formate Dehydrogenase from Candida boidinii (CbFDH) reversibly catalyzes Formate to carbon dioxide with the redox couple NAD+/NADH. There have been many studies on CbFDH-catalyzed oxidation of Formate to carbon dioxide in the presence of NAD+. On the other hand, there are few studies on the detailed mechanism of the carbon dioxide reduction to Formate catalyzed by CbFDH in the presence of NADH. In addition, it is not clear whether CbFDH can reduce carbon dioxide, bicarbonate or carbonate to Formate. In this study, Formate production with CbFDH in the presence of NADH was investigated in solutions with different ratios of carbon dioxide, bicarbonate and carbonate. The reaction rate of Formate production with CbFDH increased in proportion to the concentration of carbon dioxide in the reaction solution. On the other hand, the Formate production with CbFDH was suppressed in proportion to the concentration of bicarbonate or carbonate in the reaction solution. Thus, CbFDH was found to catalytically reduce only carbon dioxide to Formate among the three types of carbonate species.
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Formate Dehydrogenase for co2 utilization and its application
2018Co-Authors: Yutaka AmaoAbstract:Abstract Carbon dioxide, CO2 reduction and utilization for organic compounds synthesis are the potential technologies in environmental science and technology. In order to establish efficient CO2 utilization technologies, an effective catalyst for CO2 reduction and utilization is necessary. Among various catalysts, the biocatalyst is one of promising catalysts because it has excellent selectivity for the reaction and substrate. In this review, focusing on biocatalyst “Formate Dehydrogenase FDH” catalyzing CO2 reduction to formic acid, representative examples of properties, types, structure of active-site of FDH and, reaction mechanism of formic acid oxidation and CO2 reduction with FDH are outlined. A genetic engineering modified FDH and FDH immobilized various support for improving CO2 reduction catalytic activity also are introduced. Moreover, chemical and electrochemical system of CO2 reduction to formic acid with FDH, aqueous homogenous system of visible-light driven CO2 reduction to formic acid with FDH and device for visible-light driven CO2 reduction to formic acid with FDH are also introduced as an application of FDH.
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a novel electron carrier molecule based on a viologen derivative for visible light driven co2 reduction to formic acid with the system of zinc porphyrin and Formate Dehydrogenase
2017Co-Authors: Shusaku Ikeyama, Yutaka AmaoAbstract:To develop a visible-light driven conversion of CO2 to formic acid system consisting of water-soluble zinc porphyrin, an electron carrier and Formate Dehydrogenase (FDH) in the presence of triethanolamine (TEOA) as an electron donor, 1,1′-diaminoethyl-4,4′-bipyridinium salt (DAV) as a novel electron carrier with high affinity for FDH was applied.
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discovery of the reduced form of methylviologen activating Formate Dehydrogenase in the catalytic conversion of carbon dioxide to formic acid
2015Co-Authors: Yutaka Amao, Shusaku IkeyamaAbstract:The kinetic properties of the conversion of CO2 to formic acid with Formate Dehydrogenase (FDH) using dithionite-reduced methylviologen as an artificial coenzyme were studied. The catalytic product...
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photochemical synthesis of formic acid from co2 with Formate Dehydrogenase and water soluble zinc porphyrin
2004Co-Authors: Rie Miyatani, Yutaka AmaoAbstract:Abstract Photochemical synthesis of formic acid was investigated from hydrogen carbonate ion with Formate Dehydrogenase (FDH) from Saccharomyces cerevisiae and methylviologen (MV2+) photoreduction by the visible light photosensitisation of zinc tetrakis(4-methylpyridyl) porphyrin (ZnTMPyP) in the presence of triethanolamine (TEOA) as an electron donating reagent. Irradiation of a solution containing TEOA, ZnTMPyP, MV2+, sodium hydrogen carbonate and FDH with visible light resulted in formic acid synthesis. The concentration of formic acid was 62 μM after 3 h irradiation under the condition of 30 μM MV2+, 20 units FDH and pH 8.
Erwin Reisner - One of the best experts on this subject based on the ideXlab platform.
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interfacing Formate Dehydrogenase with metal oxides for the reversible electrocatalysis and solar driven reduction of carbon dioxide
2019Co-Authors: Melanie Miller, Ines Ac Pereira, William E Robinson, Ana Rita Oliveira, Nina Heidary, Nikolay Kornienko, Julien Warnan, Erwin ReisnerAbstract:The integration of enzymes with synthetic materials allows efficient electrocatalysis and production of solar fuels. Here, we couple Formate Dehydrogenase (FDH) from Desulfovibrio vulgaris Hildenborough (DvH) to metal oxides for catalytic CO2 reduction and report an in-depth study of the resulting enzyme-material interface. Protein film voltammetry (PFV) demonstrates the stable binding of FDH on metal-oxide electrodes and reveals the reversible and selective reduction of CO2 to Formate. Quartz crystal microbalance (QCM) and attenuated total reflection infrared (ATR-IR) spectroscopy confirm a high binding affinity for FDH to the TiO2 surface. Adsorption of FDH on dye-sensitized TiO2 allows for visible-light-driven CO2 reduction to Formate in the absence of a soluble redox mediator with a turnover frequency (TOF) of 11±1 s-1 . The strong coupling of the enzyme to the semiconductor gives rise to a new benchmark in the selective photoreduction of aqueous CO2 to Formate.
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photoreduction of co2 with a Formate Dehydrogenase driven by photosystem ii using a semi artificial z scheme architecture
2018Co-Authors: Katarzyna P Sokol, Ines Ac Pereira, William E Robinson, Julien Warnan, Ana R Oliveira, Marc M Nowaczyk, Adrian Ruff, Erwin ReisnerAbstract:Solar-driven coupling of water oxidation with CO2 reduction sustains life on our planet and is of high priority in contemporary energy research. Here, we report a photoelectrochemical tandem device that performs photocatalytic reduction of CO2 to Formate. We employ a semi-artificial design, which wires a W-dependent Formate Dehydrogenase (FDH) cathode to a photoanode containing the photosynthetic water oxidation enzyme, Photosystem II, via a synthetic dye with complementary light absorption. From a biological perspective, the system achieves a metabolically inaccessible pathway of light-driven CO2 fixation to Formate. From a synthetic point of view, it represents a proof-of-principle system utilizing precious-metal-free catalysts for selective CO2-to-Formate conversion using water as an electron donor. This hybrid platform demonstrates the translatability and versatility of coupling abiotic and biotic components to create challenging models for solar fuel and chemical synthesis.
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oxidation state dependent binding properties of the active site in a mo containing Formate Dehydrogenase
2017Co-Authors: William E Robinson, Arnau Bassegoda, Erwin Reisner, Judy HirstAbstract:Molybdenum-containing Formate Dehydrogenase H from Escherichia coli (EcFDH-H) is a powerful model system for studies of the reversible reduction of CO2 to Formate. However, the mechanism of FDH catalysis is currently under debate, and whether the primary Mo coordination sphere remains saturated or one of the ligands dissociates to allow direct substrate binding during turnover is disputed. Herein, we describe how oxidation-state-dependent changes at the active site alter its inhibitor binding properties. Using protein film electrochemistry, we show that Formate oxidation by EcFDH-H is inhibited strongly and competitively by N3–, OCN–, SCN–, NO2–, and NO3–, whereas CO2 reduction is inhibited only weakly and not competitively. During catalysis, the Mo center cycles between the formal Mo(VI)═S and Mo(IV)—SH states, and by modeling chronoamperometry data recorded at different potentials and substrate and inhibitor concentrations, we demonstrate that both Formate oxidation and CO2 reduction are inhibited by se...
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reversible interconversion of co2 and Formate by a molybdenum containing Formate Dehydrogenase
2014Co-Authors: Arnau Bassegoda, Christopher Madden, David W Wakerley, Erwin Reisner, Judy HirstAbstract:CO2 and Formate are rapidly, selectively, and efficiently interconverted by tungsten-containing Formate Dehydrogenases that surpass current synthetic catalysts. However, their mechanism of catalysis is unknown, and no tractable system is available for study. Here, we describe the catalytic properties of the molybdenum-containing Formate Dehydrogenase H from the model organism Escherichia coli (EcFDH-H). We use protein film voltammetry to demonstrate that EcFDH-H is a highly active, reversible electrocatalyst. In each voltammogram a single point of zero net current denotes the CO2 reduction potential that varies with pH according to the Nernst equation. By quantifying Formate production we show that electrocatalytic CO2 reduction is specific. Our results reveal the capabilities of a Mo-containing catalyst for reversible CO2 reduction and establish EcFDH-H as an attractive model system for mechanistic investigations and a template for the development of synthetic catalysts.