Reversible Interconversion of CO<sub>2</sub> and Formate by a Molybdenum-Containing Formate Dehydrogenase

Bassegoda, Arnau; Madden, Christopher; Wakerley, David W.; Reisner, Erwin; Hirst, Judy

doi:10.1021/ja508647u

Cited by 215 publications

(208 citation statements)

References 28 publications

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“…This kinetic behavior is not exclusive to the Dd FDH and initial lag phases were also described with, e.g., Syntrophobacter fumaroxidans W-FDH1. 14 The lag phase extent was found to be dependent on the enzyme and formate concentrations, with shorter lag phases being obtained with higher enzyme or formate concentrations (Fig. 2, curves a-e).…”

Section: Transients During Fdh-catalyzed Formate Oxidation (Initial Lmentioning

confidence: 88%

Reduction of Carbon Dioxide by a Molybdenum-Containing Formate Dehydrogenase: A Kinetic and Mechanistic Study

et al. 2016

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Carbon dioxide accumulation is a major concern for the ecosystems, but its abundance and low cost make it an interesting source for the production of chemical feedstocks and fuels. However, the thermodynamic and kinetic stability of the carbon dioxide molecule makes its activation a challenging task. Studying the chemistry used by nature to functionalize carbon dioxide should be helpful for the development of new efficient (bio)catalysts for atmospheric carbon dioxide utilization. In this work, the ability of Desulfovibrio desulfuricans formate dehydrogenase (Dd FDH) to reduce carbon dioxide was kinetically and mechanistically characterized. The Dd FDH is suggested to be purified in an inactive form that has to be activated through a reduction-dependent mechanism. A kinetic model of a hysteretic enzyme is proposed to interpret and predict the progress curves of the Dd FDH-catalyzed reactions (initial lag phase and subsequent faster phase). Once activated, Dd FDH is able to efficiently catalyze, not only the formate oxidation (k cat of 543 s–1, K m of 57.1 μM), but also the carbon dioxide reduction (k cat of 46.6 s–1, K m of 15.7 μM), in an overall reaction that is thermodynamically and kinetically reversible. Noteworthy, both Dd FDH-catalyzed formate oxidation and carbon dioxide reduction are completely inactivated by cyanide. Current FDH reaction mechanistic proposals are discussed and a different mechanism is here suggested: formate oxidation and carbon dioxide reduction are proposed to proceed through hydride transfer and the sulfo group of the oxidized and reduced molybdenum center, Mo6+S and Mo4+-SH, are suggested to be the direct hydride acceptor and donor, respectively.

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Section: Transients During Fdh-catalyzed Formate Oxidation (Initial Lmentioning

confidence: 88%

Reduction of Carbon Dioxide by a Molybdenum-Containing Formate Dehydrogenase: A Kinetic and Mechanistic Study

et al. 2016

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“…Mo-FDH from E. coli is the best characterized metal dependent FDH, and easy to produce in a stable, reduced form yielding significant advantages for the EFC field. 10,13 Therefore, future work will focus on developing materials strategies for improving the performance of the Mo-FDH bioanodes for fuel cell applications.…”

Section: Discussionmentioning

confidence: 99%

“…Work by Bassegoda et al 13 and Robinson et al 15 reported the reversible interconversion of CO 2 and formate without using any mediator with the FDH adsorbed on graphite-epoxy rotating disk working electrode. While the direct electrochemical approach adopted by the authors is well-suited to study finer thermodynamic and kinetic properties of metalloenzymes (i.e., FDH), mediated electrochemical approaches facilitated by redox polymers (BPV-LPEI) are better suited for application in devices where increased enzyme loading and current densities can be realized at an electrode surface.…”

Section: -29mentioning

confidence: 99%

Molybdenum-Dependent Formate Dehydrogenase for Formate Bioelectrocatalysis in a Formate/O₂Enzymatic Fuel Cell

Şahin

Cai

Milton

et al. 2018

J. Electrochem. Soc.

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Formate/formic acid has been an intriguing fuel for fuel cells and biofuel cells over the last two decades. The common challenge with formate/formic acid biofuel cells has been the use of NAD-dependent formate dehydrogenase, which requires a diffusive cofactor and problematic cofactor regeneration systems. In this paper, we explore the bioelectro-oxidation of formate using Mo-containing formate dehydrogenase from E. coli (Mo-FDH) for development of a new HCOO − /O 2 enzymatic fuel cell (EFC). Mo-FDH was coupled with a benzylpropylviologen-based linear polyethylenimine (BPV-LPEI) redox polymer to fabricate a formate bioanode and laccase incorporated with anthracene-modified multi-walled carbon nanotubes (Ac-MWCNTs) to promote direct electron transfer was used for the O 2 biocathode. The resulting Mo-FDH/laccase EFC has an open-circuit potential of 1.28 ± 0.05 V, with corresponding maximum current and power densities of 17 ± 7 μA cm −2 and 18 ± 6 μW cm −2 , respectively. This FDH contains a molybdenum-coordinated selenocysteine (SeCys) residue, essential for catalytic activity, and two molybdopterin guanine dinucleotides (MGDs), with just one [4Fe-4S] cluster for electron transfer to and from the active site.13,14 Bassegoda et al. 13and Robinson et al. 15 showed that Mo-FDH from E.coli adsorbed on the surface of a graphite-epoxy rotating disk working electrode can catalyze the reversible interconversion of CO 2 and formate with direct electron transfer at ∼ −0.4 V vs. SHE (pH 6.8).16 However, the current densities of DET electrodes are lower than what is necessary for biofuel cell applications.In this study, we constructed a MET-type bioanode with Mo-FDH by employing a viologen-based redox polymer (benzylpropylviologen-modified linear poly(ethylenimine), BPV-LPEI) which is able to facilitate the bioelectrocatalytic oxidation of formate at low potentials. While a DET approach could theoretically offer improved open-circuit potentials of a resulting EFC, the current densities obtained from DET-based bioelectrodes is almost always dwarfed by those obtained from appropriately designed MET bioanodes, due to a combination of increased enzyme loading, the ability of redox polymers to undergo "self-exchange" (creating an expanded 3D network of electronically-connected yet immobilized enzyme) and increased interfacial electron transfer rates (k ET ) which are commonly poor for DET-based bioelectrodes.17,18 Therefore, we investigated a redox polymer with low oxidation potentials to ensure near theoretical OCPs for the EFC. Several different mechanisms for FDH-catalyzed formate oxidation have been proposed. 5,9,14,19,20 It was shown by Maia et al. that FDH requires an activation step by its reduction using viologen or an artificial reducer.9 Robinson et al. have proposed a reductive activation to create a stabilized form of the SeCys dissociated state and it was mentioned that this state is also possible upon incubation of the protein with reducing agents (such as sodium dithionite (DT)). 15,19 Thus, we evaluated the activat...

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“…Two classes of FDHs have been discovered thus far: metal-independent nicotinamide adenine dinucleotide (NADH)-based FDHs [1,2] and metal-dependent molybdenum (Mo)-based or tungsten (W)-based FDHs [3][4][5]. We had earlier created several NADPH-dependent type FDHs and combined them with photosystem I (PSI), ferredoxin, and ferredoxin-NADP + -reductase to establish an artificial light-driven carbon dioxide reduction system (Fig.…”

Section: Introductionmentioning

confidence: 99%

“…The dimer of FdoG and FdoH is directed toward the cytoplasmic side, and with FdoH anchored to the periplasmic membrane and the FdoI subunit by a C-terminal hydrophobic domain [16]. Among these three metal-dependent FDHs from E. coli, EcFDH-H has been well-studied due to its structural simplicity and solubility [3], but it is intractable for practical applications due to its extremely high oxygen-sensitivity.…”

Section: Introductionmentioning

confidence: 99%

Expression systems for soluble metal-dependent formate dehydrogenase

Ihara

Kawano

Fujiwara

et al. 2015

Journal of Photochemistry and Photobiology A: Chemistry

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Molybdenum or tungsten-dependent formate dehydrogenases (FDH) can reduce carbon dioxide (CO 2 ) to formate under ordinary conditions, and therefore, are considered promising catalysts for CO 2 fixation. However, to our knowledge, no study on the modification of metal-dependent FDHs has been published, likely because of a lack of convenient expression systems for the recombinant enzymes. We attempted to establish an methodology for the preparation and modification of soluble oxygen-tolerant metaldependent FDHs, for the following three strategies: (1) Escherichia coli FDH is converted from a membrane bound protein into a soluble protein by deleting the C-terminal membrane-anchor of small subunit (FdoH) and the whole membrane subunit (FdoI) and expressed homologously in E. coli; (2) originally soluble FDHs from Desulfovibrio are expressed heterologously in E. coli; and (3) Desulfovibrio FDHs are genetically engineered by the homologous gene recombination method and expressed homologously in Desulfovibrio. We successfully established the expression systems for (1) and (3), and succeeded in purification of the soluble FDHs with a tag by using affinity columns.

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Reversible Interconversion of CO₂ and Formate by a Molybdenum-Containing Formate Dehydrogenase

Cited by 215 publications

References 28 publications

Reduction of Carbon Dioxide by a Molybdenum-Containing Formate Dehydrogenase: A Kinetic and Mechanistic Study

Reduction of Carbon Dioxide by a Molybdenum-Containing Formate Dehydrogenase: A Kinetic and Mechanistic Study

Molybdenum-Dependent Formate Dehydrogenase for Formate Bioelectrocatalysis in a Formate/O₂Enzymatic Fuel Cell

Expression systems for soluble metal-dependent formate dehydrogenase

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Reversible Interconversion of CO2 and Formate by a Molybdenum-Containing Formate Dehydrogenase

Cited by 215 publications

References 28 publications

Reduction of Carbon Dioxide by a Molybdenum-Containing Formate Dehydrogenase: A Kinetic and Mechanistic Study

Reduction of Carbon Dioxide by a Molybdenum-Containing Formate Dehydrogenase: A Kinetic and Mechanistic Study

Molybdenum-Dependent Formate Dehydrogenase for Formate Bioelectrocatalysis in a Formate/O2Enzymatic Fuel Cell

Expression systems for soluble metal-dependent formate dehydrogenase

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Reversible Interconversion of CO₂ and Formate by a Molybdenum-Containing Formate Dehydrogenase

Molybdenum-Dependent Formate Dehydrogenase for Formate Bioelectrocatalysis in a Formate/O₂Enzymatic Fuel Cell