An unprecedented function for a tungsten-containing oxidoreductase

Mathew, Liju; Haja, Dominik K.; Pritchett, Clayton; McCormick, Winston; Zeineddine, Robbie; Fontenot, Leo; Rivera, Mario; Glushka, John; Adams, Michael W. W.; Lanzilotta, William N.

doi:10.1007/s00775-022-01965-0

Cited by 5 publications

(11 citation statements)

References 38 publications

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“…These residues are conserved and appear similarly used as in AOR Pf, whereas they are not conserved in FOR, which exhibits some changes in the W-co binding motifs. No further metal ions were identified in close proximity to any of the pterins in AOR Aa , in contrast to the available structures AOR Pf , FOR Pf , and WOR5 that show sodium (8), calcium (9), or two additional magnesium ions (10), respectively.…”

Section: Aorb Binds a W Cofactorcontrasting

confidence: 75%

A bacterial tungsten-containing aldehyde oxidoreductase forms an enzymatic decorated protein nanowire

et al. 2023

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Aldehyde oxidoreductases (AORs) are tungsten enzymes catalyzing the oxidation of many different aldehydes to the corresponding carboxylic acids. In contrast to other known AORs, the enzyme from the denitrifying betaproteobacterium Aromatoleum aromaticum (AOR Aa ) consists of three different subunits (AorABC) and uses nicotinamide adenine dinucleotide (NAD) as an electron acceptor. Here, we reveal that the enzyme forms filaments of repeating AorAB protomers that are capped by a single NAD-binding AorC subunit, based on solving its structure via cryo–electron microscopy. The polyferredoxin-like subunit AorA oligomerizes to an electron-conducting nanowire that is decorated with enzymatically active and W-cofactor (W-co) containing AorB subunits. Our structure further reveals the binding mode of the native substrate benzoate in the AorB active site. This, together with quantum mechanics:molecular mechanics (QM:MM)–based modeling for the coordination of the W-co, enables formulation of a hypothetical catalytic mechanism that paves the way to further engineering for applications in synthetic biology and biotechnology.

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Section: Aorb Binds a W Cofactorcontrasting

confidence: 75%

A bacterial tungsten-containing aldehyde oxidoreductase forms an enzymatic decorated protein nanowire

et al. 2023

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“…These residues are conserved and appear similarly employed as in AOR Pf whereas they are not conserved in FOR, which exhibits some changes in the W-co binding motifs. No further metal ions were identified in close proximity to any of the pterins in AOR Aa , in contrast to the available structures AOR Pf , FOR Pf and WOR5 that show sodium ( 8 ), calcium ( 9 ) or two additional magnesium ions ( 10 ), respectively.…”

Section: Resultscontrasting

confidence: 70%

“…These residues are conserved and appear similarly employed as in AORPf, whereas they are not conserved in FOR, which exhibits some changes in the W-co binding motifs. No further metal ions were identified in close proximity to any of the pterins in AORAa, in contrast to the available structures AORPf, FORPf and WOR5 that show sodium (8), calcium (9) or two additional magnesium ions (10), respectively. The highly conserved residues His 484 and Glu 331 (79% and 82% conserved throughout the AOR family, respectively) are located close to the tungsten and therefore are possible candidates for being catalytically active residues.…”

Section: Cofactor Geometry In Aorbcontrasting

confidence: 71%

“…The short distances below 15 Å between all redox cofactors are expected to support rapid electron transfer via quantum tunnelling within microseconds. The presence of AorA or similar FeS-cluster-containing subunits has only been observed in a few clades of the AOR family, such as the AOR Aa , GOR, WOR5, and Bam clades, while the other characterized members (including AOR Pf and FOR) do not contain such a subunit ( 1, 10, 21 ). Moreover, none of the other known tungsten enzymes with such a subunit form oligomers.…”

Section: Resultsmentioning

confidence: 99%

“…Although they can perform such highly sophisticated redox reactions, our structural and mechanistic understanding of AOR enzymes has been sparse. Currently, the only structurally characterised tungsten aldehyde oxidoreductases come from the hyperthermophilic archeon Pyrococcus furiosus: namely AOR ( sensu stricto ) ( 8 ), formaldehyde oxidoreductase (FOR) ( 9 ), and tungsten (W)-containing oxidoreductase 5 (WOR5) ( 10 ). Those structures revealed the fold of the AOR catalytic subunit and general structure of the metallopterin and tungsten cofactor.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

A bacterial tungsten-containing aldehyde oxidoreductase forms an enzymatic decorated protein nanowire

Winiarska

Ramírez-Amador

Hege

et al. 2023

Preprint

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Aldehyde oxidoreductases (AOR) are tungsten enzymes catalysing the oxidation of many different aldehydes to the corresponding carboxylic acids. In contrast to other known AORs, the enzyme from the denitrifying betaproteobacteriumAromatoleum aromaticum(AORAa) consists of three different subunits (AorABC) and utilizes NAD as electron acceptor. Here we reveal that the enzyme forms filaments of repeating AorAB protomers which are capped by a single NAD-binding AorC subunit, based on solving its structure via cryo-electron microscopy. The polyferredoxin-like subunit AorA oligomerizes to an electron-conducting nanowire that is decorated with enzymatically active and W-cofactor (W-co) containing AorB subunits. Our structure further reveals the binding mode of the native substrate benzoate in the AorB active site. This, together with QM:MM-based modelling for the coordination of the W-co, enables formulation of catalytic mechanism hypothesis that paves the way for further engineering of AOR for applications in synthetic biology and biotechnology.

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Carbon monoxide-driven bioethanol production operates via a tungsten-dependent catalyst

Lemaire,

Belhamri,

Schevchenko

et al. 2024

Preprint

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Microbial alcohol production from waste gasses is a game changer for sustainable carbon cycling and remediation. While the biotechnological process employingClostridium autoethanogenumto transform syngas (H2/CO2/CO) is blooming, the reactions involved in ethanol biosynthesis remain to be demonstrated. Here, we experimentally showed that ethanol production initiates via a tungsten-dependent aldehyde:ferredoxin oxidoreductase (AFOR), which reduces acetate to acetaldehyde. Such an unfavourable reaction has often been considered unsuitable for a biological process. To answer this riddle, we demonstrated that the thermodynamic pull of CO-oxidation and ethanol synthesis is crucial for triggering acetate reduction. The experimental setup performed with native CO-dehydrogenase and AFOR highlighted the key role of ferredoxin in stimulating the activity of both metalloenzymes and electron shuttling. The crystal structure of holo AFOR refined to 1.59-Å resolution, together with its biochemical characterisation, provides new insights into the reaction mechanism and the specificities of this enzyme fundamental to sustainable biofuel production.

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An unprecedented function for a tungsten-containing oxidoreductase

Cited by 5 publications

References 38 publications

A bacterial tungsten-containing aldehyde oxidoreductase forms an enzymatic decorated protein nanowire

A bacterial tungsten-containing aldehyde oxidoreductase forms an enzymatic decorated protein nanowire

A bacterial tungsten-containing aldehyde oxidoreductase forms an enzymatic decorated protein nanowire

Carbon monoxide-driven bioethanol production operates via a tungsten-dependent catalyst

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