A synthetic pathway for the fixation of carbon dioxide in vitro

Schwander, Thomas; Borzyskowski, Lennart Schada von; Burgener, Simon; Cortina, Niña Socorro; Erb, Tobias J.

doi:10.1126/science.aah5237

Cited by 563 publications

(559 citation statements)

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“…The recently presented CETCH cycle of Schwander et al . is pioneering in being the first synthetic CO 2 fixation cycle. The cycle, however, does not include PFOR but uses crotonyl‐CoA carboxylase/reductase as CO 2 ‐fixing enzyme to yield glyoxylate as end product.…”

Section: Resultsmentioning

confidence: 99%

New light on ancient enzymes – in vitro CO₂ Fixation by Pyruvate Synthase of Desulfovibrio africanus and Sulfolobus acidocaldarius

et al. 2019

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Two variants of the enzyme family pyruvate:ferredoxin oxidoreductase (PFOR), derived from the anaerobic sulfate‐reducing bacterium Desulfovibrio africanus and the extremophilic crenarchaeon Sulfolobus acidocaldarius, respectively, were evaluated for their capacity to fixate CO2 in vitro. PFOR reversibly catalyzes the conversion of acetyl‐CoA and CO2 to pyruvate using ferredoxin as redox partner. The oxidative decarboxylation of pyruvate is thermodynamically strongly favored, and most previous studies only considered the oxidative direction of the enzyme. To assay the pyruvate synthase function of PFOR during reductive carboxylation of acetyl‐CoA is more challenging and requires to maintain the reaction far from equilibrium. For this purpose, a biochemical assay was established where low‐potential electrons were introduced by photochemical reduction of EDTA/deazaflavin and the generated pyruvate was trapped by chemical derivatization with semicarbazide. The product of CO2 fixation could be detected as pyruvate semicarbazone by HPLC‐MS. In a combinatorial approach, both PFORs were tested with ferredoxins from different sources. The pyruvate semicarbazone product could be detected with low‐potential ferredoxins of the green sulfur bacterium Chlorobium tepidum and of S. acidocaldarius whereas CO2 fixation was not supported by the native ferredoxin of D. africanus. Methylviologen as an artificial electron carrier also allowed CO2 fixation. For both enzymes, the results are the first demonstration of CO2 fixation in vitro. Both enzymes exhibited high stability in the presence of oxygen during purification and storage. In conclusion, the employed PFOR enzymes in combination with non‐native ferredoxin cofactors might be promising candidates for further incorporation in biocatalytic CO2 conversion. Enzymes EC1.2.7.1. Pyruvate:Ferredoxin Oxidoreductase

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Section: Resultsmentioning

confidence: 99%

New light on ancient enzymes – in vitro CO₂ Fixation by Pyruvate Synthase of Desulfovibrio africanus and Sulfolobus acidocaldarius

et al. 2019

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“…Recent examples include replicating systems, [4,5] transient self-assembly, [6][7][8] active colloids, [9] and many others. [15][16][17] However,the development of enzymatic reaction networks [18][19][20][21] has been slow.Incontrast to most DNA-based networks, [15,16] where interactions can be "programmed" based on base-pair interactions,t he reaction kinetics of enzymatic reactions cannot be predicted apriori. [15][16][17] However,the development of enzymatic reaction networks [18][19][20][21] has been slow.Incontrast to most DNA-based networks, [15,16] where interactions can be "programmed" based on base-pair interactions,t he reaction kinetics of enzymatic reactions cannot be predicted apriori.…”

Section: Chemicalreactionnetworkdrivemanyofthekeyprocessesmentioning

confidence: 99%

Modular Design of Small Enzymatic Reaction Networks Based on Reversible and Cleavable Inhibitors

Regueiro¹,

Jakštaitė²,

Hollander³

et al. 2019

Angew Chem Int Ed

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Systems chemistry aims to mimic the functional behavior of living systems by constructing chemical reaction networks with well-defined dynamic properties.E nzymes can play ak ey role in such networks,b ut there is currently no general and scalable route to the design and construction of enzymatic reaction networks.H ere,w ei ntroduce reversible, cleavable peptide inhibitors that can link proteolytic enzymatic activity into simple network motifs.Asaproof-of-principle,we show auto-activation topologies producing sigmoidal responses in enzymatic activity,e xplore cross-talk in minimal systems,d esign as imple enzymatic cascade,a nd introduce non-inhibiting phosphorylated peptides that can be activated using aphosphatase.

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“…References are color‐coded green; purple; and yellow . B) Simplified representation of the first synthetic in vitro pathway for CO 2 fixation (CETCH cycle).…”

Section: Case Studies Of Pathway Design and Engineeringmentioning

confidence: 99%

Expanding Metabolic Capabilities Using Novel Pathway Designs: Computational Tools and Case Studies

Saa

Cortés

López

et al. 2019

Biotechnology Journal

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Design and selection of efficient metabolic pathways is critical for the success of metabolic engineering endeavors. Convenient pathways should not only produce the target metabolite in high yields but also are required to be thermodynamically feasible under production conditions, and to prefer efficient enzymes. To support the design and selection of such pathways, different computational approaches have been proposed for exploring the feasible pathway space under many of the above constraints. In this review, an overview of recent constraint‐based optimization frameworks for metabolic pathway prediction, as well as relevant pathway engineering case studies that highlight the importance of rational metabolic designs is presented. Despite the availability and suitability of in silico design tools for metabolic pathway engineering, scarce—although increasing—application of computational outcomes is found. Finally, challenges and limitations hindering the broad adoption and successful application of these tools in metabolic engineering projects are discussed.

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A synthetic pathway for the fixation of carbon dioxide in vitro

Cited by 563 publications

References 59 publications

New light on ancient enzymes – in vitro CO₂ Fixation by Pyruvate Synthase of Desulfovibrio africanus and Sulfolobus acidocaldarius

New light on ancient enzymes – in vitro CO₂ Fixation by Pyruvate Synthase of Desulfovibrio africanus and Sulfolobus acidocaldarius

Modular Design of Small Enzymatic Reaction Networks Based on Reversible and Cleavable Inhibitors

Expanding Metabolic Capabilities Using Novel Pathway Designs: Computational Tools and Case Studies

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A synthetic pathway for the fixation of carbon dioxide in vitro

Cited by 563 publications

References 59 publications

New light on ancient enzymes – in vitro CO2 Fixation by Pyruvate Synthase of Desulfovibrio africanus and Sulfolobus acidocaldarius

New light on ancient enzymes – in vitro CO2 Fixation by Pyruvate Synthase of Desulfovibrio africanus and Sulfolobus acidocaldarius

Modular Design of Small Enzymatic Reaction Networks Based on Reversible and Cleavable Inhibitors

Expanding Metabolic Capabilities Using Novel Pathway Designs: Computational Tools and Case Studies

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New light on ancient enzymes – in vitro CO₂ Fixation by Pyruvate Synthase of Desulfovibrio africanus and Sulfolobus acidocaldarius

New light on ancient enzymes – in vitro CO₂ Fixation by Pyruvate Synthase of Desulfovibrio africanus and Sulfolobus acidocaldarius