In Vivo Assimilation of One-Carbon via a Synthetic Reductive Glycine Pathway in Escherichia coli

Yishai, Oren; Bouzon, Madeleine; Döring, Volker; Bar‐Even, Arren

doi:10.1021/acssynbio.8b00131

Cited by 137 publications

(104 citation statements)

References 38 publications

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“…By using NCD‐linked FDH* and ME*, we demonstrated successful reductive carboxylation of pyruvate driven by formate and NCD in vitro and in vivo. Recently, formate has been applied to support cell growth of E. coli and Saccharomyces cerevisiae , or to provide reducing power for autotrophic growth on CO 2 by engineered E. coli . With FDH* and other NCD‐linked enzymes, it may provide fascinating opportunities to selectively deliver reducing power, together with CO 2 if an NCD‐linked enzyme such as ME* is involved, for valuable metabolite production, which fits well with the idea of the formate bio‐economy .…”

Section: Methodsmentioning

confidence: 99%

Non‐natural Cofactor and Formate‐Driven Reductive Carboxylation of Pyruvate

et al. 2020

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A non‐natural cofactor and formate driven system for reductive carboxylation of pyruvate is presented. A formate dehydrogenase (FDH) mutant, FDH*, that favors a non‐natural redox cofactor, nicotinamide cytosine dinucleotide (NCD), for generation of a dedicated reducing equivalent at the expense of formate were acquired. By coupling FDH* and NCD‐dependent malic enzyme (ME*), the successful utilization of formate is demonstrated as both CO2 source and electron donor for reductive carboxylation of pyruvate with a perfect stoichiometry between formate and malate. When 13C‐isotope‐labeled formate was used in in vitro trials, up to 53 % of malate had labeled carbon atom. Upon expression of FDH* and ME* in the model host E. coli, the engineered strain produced more malate in the presence of formate and NCD. This work provides an alternative and atom‐economic strategy for CO2 fixation where formate is used in lieu of CO2 and offers dedicated reducing power.

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Section: Methodsmentioning

confidence: 99%

Non‐natural Cofactor and Formate‐Driven Reductive Carboxylation of Pyruvate

et al. 2020

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“…This means that most carbon incorporated into metabolism has to come through enzymatic routes and does not fully leverage the advantages of electrochemical reduction of CO2 to formate. However, recent advances in computational design of synthetic metabolic pathways have yielded several designs that do not rely upon any enzymatic fixation of CO2 [127,128]. The most promising are shown in Table 5.…”

Section: Formate and Formic Acidmentioning

confidence: 99%

“…Representative set of pathways for processing partially reduced carbon. References [127,128,153,[171][172][173][174][175] The electrode overpotential is estimated by subtracting the estimated applied electrode potential from the assumed electron acceptor potential at pH 7 (Mtr EET Complex, E = -0.1 V vs SHE; H2, E = -0.42 V vs. SHE; Formate, E = -0.43 V vs. SHE). References [43,45,63,[139][140][141][142][143][144] Table 5.…”

Section: Figures and Tablesmentioning

confidence: 99%

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Electrical Energy Storage with Engineered Biological Systems

Salimijazi

Parra²,

Barstow

2019

Preprint

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The availability of renewable energy technologies is increasing dramatically across the globe thanks to their growing maturity. However, large scale electrical energy storage and retrieval will almost certainly be a required in order to raise the penetration of renewable sources into the grid. No present energy storage technology has the perfect combination of high power and energy density, low financial and environmental cost, lack of site restrictions, long cycle and calendar lifespan, easy materials availability, and fast response time. Engineered electroactive microbes could address many of the limitations of current energy storage technologies by enabling rewired carbon fixation, a process that spatially separates reactions that are normally carried out together in a photosynthetic cell and replaces the least efficient with non-biological equivalents. If successful, this could allow storage of renewable electricity through electrochemical or enzymatic fixation of carbon dioxide and subsequent storage as carbon-based energy storage molecules including hydrocarbon and non-volatile polymers at high efficiency. In this article we compile performance data on biological and non-biological component choices for rewired carbon fixation systems and identify pressing research and engineering challenges.

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“…The GCS reaction cycle is fully reversible and can produce glycine from 5,10-methylenetetrahydrofolate, CO 2 , NH 3 and NADH (Kawasaki et al , 1966, Freudenberg and Andreesen, 1989). Hence, the GCS is being used in synthetic biology as a key component of the ‘reductive glycine pathway’ of formate and CO 2 assimilation in tailor-made microbial organisms for sustainable bioproduction (discussed in Bar-Even, 2016, Yishai et al , 2018).…”

Section: Introductionmentioning

confidence: 99%

Stoichiometry of two plant glycine decarboxylase complexes and comparison with a cyanobacterial glycine cleavage system

Wittmiß

Mikkat

Hagemann

et al. 2020

Preprint

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ABSTRACTThe multienzyme glycine cleavage system (GCS) converts glycine and tetrahydrofolate to the one-carbon compound 5,10-methylenetetrahydrofolate, which is of vital importance for most if not all organisms. Photorespiring plant mitochondria contain very high levels of GCS proteins organised as a fragile glycine decarboxylase complex (GDC). The aim of this study is to provide mass spectrometry-based stoichiometric data for the plant leaf GDC and examine whether complex formation could be a general property of the GCS in photosynthesizing organisms. The molar ratios of the leaf GDC component proteins are 1L2-4P2-8T-26H and 1L2-4P2-8T-20H for pea and Arabidopsis, respectively, as determined by mass spectrometry. The minimum mass of the plant leaf GDC ranges from 1,550-1,650 kDa, which is larger than previously assumed. The Arabidopsis GDC contains four times more of the isoforms GCS-P1 and GCS-L1 in comparison with GCS-P2 and GCS-L2, respectively, whereas the H-isoproteins GCS-H1 and GCS-H3 are fully redundant as indicated by their about equal amounts. Isoform GCS-H2 is not present in leaf mitochondria. In the cyanobacterium Synechocystis sp. PCC 6803, GCS proteins are present at low concentration but above the complex formation threshold reported for pea leaf GDC. Indeed, formation of a cyanobacterial GDC from the individual recombinant GCS proteins in vitro could be demonstrated. Presence and metabolic significance of a Synechocystis GDC in vivo remain to be examined but could involve multimers of the GCS H-protein that dynamically crosslink the three GCS enzyme proteins, facilitating glycine metabolism by the formation of multienzyme metabolic complexes.

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In Vivo Assimilation of One-Carbon via a Synthetic Reductive Glycine Pathway in Escherichia coli

Cited by 137 publications

References 38 publications

Non‐natural Cofactor and Formate‐Driven Reductive Carboxylation of Pyruvate

Non‐natural Cofactor and Formate‐Driven Reductive Carboxylation of Pyruvate

Electrical Energy Storage with Engineered Biological Systems

Stoichiometry of two plant glycine decarboxylase complexes and comparison with a cyanobacterial glycine cleavage system

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