Exceeding the theoretical fermentation yield in mixotrophic Rubisco-based engineered Escherichia coli

Tseng, I-Ting; Chen, Ching-Hsun; Shen, Zhi-Xuan; Yang, Cheng‐Han; Li, Siyu

doi:10.1016/j.ymben.2018.04.018

Cited by 38 publications

(29 citation statements)

References 32 publications

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“…The GED shunt might have biotechnological applications on its own. Previous studies have demonstrated that co-assimilation of CO 2 can increase production yields from common feedstocks such as sugars 59 – 61 . This is attributed to the fact that the biosynthesis of certain compounds from sugars results in the production of excess reducing power, which can be utilized to fix CO 2 and thereby generate more product.…”

Section: Discussionmentioning

confidence: 99%

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Awakening a latent carbon fixation cycle in Escherichia coli

et al. 2020

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Carbon fixation is one of the most important biochemical processes. Most natural carbon fixation pathways are thought to have emerged from enzymes that originally performed other metabolic tasks. Can we recreate the emergence of a carbon fixation pathway in a heterotrophic host by recruiting only endogenous enzymes? In this study, we address this question by systematically analyzing possible carbon fixation pathways composed only of Escherichia coli native enzymes. We identify the GED (Gnd–Entner–Doudoroff) cycle as the simplest pathway that can operate with high thermodynamic driving force. This autocatalytic route is based on reductive carboxylation of ribulose 5-phosphate (Ru5P) by 6-phosphogluconate dehydrogenase (Gnd), followed by reactions of the Entner–Doudoroff pathway, gluconeogenesis, and the pentose phosphate pathway. We demonstrate the in vivo feasibility of this new-to-nature pathway by constructing E. coli gene deletion strains whose growth on pentose sugars depends on the GED shunt, a linear variant of the GED cycle which does not require the regeneration of Ru5P. Several metabolic adaptations, most importantly the increased production of NADPH, assist in establishing sufficiently high flux to sustain this growth. Our study exemplifies a trajectory for the emergence of carbon fixation in a heterotrophic organism and demonstrates a synthetic pathway of biotechnological interest.

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Section: Discussionmentioning

confidence: 99%

“…4 ). While the RuBP shunt—a linear version of the RuBP cycle, which channels Ru5P via Rubisco—can also increase the fermentative yield of some products 59 – 61 , the GED shunt always outperforms it due to a lower ATP requirement (Fig. 5 and Supplementary Fig.…”

Section: Discussionmentioning

confidence: 99%

Awakening a latent carbon fixation cycle in Escherichia coli

et al. 2020

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“…While heterotrophic organisms are not naturally able to assimilate carbon dioxide, an attractive and complementary strategy to those developed elsewhere to capture CO 2 is to equip these organisms with the two key Calvin cycle enzymes in order to capture CO 2 produced during sugar fermentation, and meanwhile use it as an electron acceptor for NADH reoxidation. This dual function has been developed in yeast and E. coli and was shown to increase C2-products such as ethanol (Papapetridis et al, 2018;Tseng et al, 2018). The high concentration of carbon dioxide that prevails in industrial fermentations and their usual anaerobic to micro-aeration conditions are factors that should support the proper functioning of this capture system since RuBisCO has low affinity to CO 2 and its carboxylation activity is antagonized by oxygen.…”

Section: Discussionmentioning

confidence: 99%

Synthetic Biology Applied to Carbon Conservative and Carbon Dioxide Recycling Pathways

François

Lachaux²,

Morin

2020

Front. Bioeng. Biotechnol.

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The global warming conjugated with our reliance to petrol derived processes and products have raised strong concern about the future of our planet, asking urgently to find sustainable substitute solutions to decrease this reliance and annihilate this climate change mainly due to excess of CO 2 emission. In this regard, the exploitation of microorganisms as microbial cell factories able to convert non-edible but renewable carbon sources into biofuels and commodity chemicals appears as an attractive solution. However, there is still a long way to go to make this solution economically viable and to introduce the use of microorganisms as one of the motor of the forthcoming bio-based economy. In this review, we address a scientific issue that must be challenged in order to improve the value of microbial organisms as cell factories. This issue is related to the capability of microbial systems to optimize carbon conservation during their metabolic processes. This initiative, which can be addressed nowadays using the advances in Synthetic Biology, should lead to an increase in products yield per carbon assimilated which is a key performance indice in biotechnological processes, as well as to indirectly contribute to a reduction of CO 2 emission.

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“…S5, and Methods). While the RuBP shunta linear version of the RuBP cycle, which channels Ru5P via Rubiscocan also increase fermentative yield of some products [60][61][62] , the GED shunt always outperforms it due to a lower ATP requirement ( Fig. 5 and Supplementary Fig.…”

Section: Discussionmentioning

confidence: 99%

Awakening a latent carbon fixation cycle inEscherichia coli

Satanowski

Dronsella

Noor

et al. 2020

Preprint

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Carbon fixation is one of the most important biochemical processes. Most natural carbon fixation pathways are thought to have emerged from enzymes that originally performed other metabolic tasks. Can we recreate the emergence of a carbon fixation pathway in a heterotrophic host by recruiting only endogenous enzymes? In this study, we address this question by systematically analyzing possible carbon fixation pathways composed only of Escherichia coli native enzymes. We identify the GED (Gnd-Entner-Doudoroff) cycle as the simplest pathway that can operate with high thermodynamic driving force. This autocatalytic route is based on reductive carboxylation of ribulose 5-phosphate (Ru5P) by 6-phosphogluconate dehydrogenase (Gnd), followed by reactions of the Entner-Doudoroff pathway, gluconeogenesis, and the pentose phosphate pathway. We demonstrate the in vivo feasibility of this new-to-nature pathway by constructing E. coli gene deletion strains whose growth on pentose sugars depends on the GED shunt, a linear variant of the GED cycle which does not require the regeneration of Ru5P. Several metabolic adaptations, most importantly the increased production of NADPH, assist in establishing sufficiently high flux to sustain this growth. Our study exemplifies a trajectory for the emergence of carbon fixation in a heterotrophic organism and demonstrates a synthetic pathway of biotechnological interest.

show abstract

Exceeding the theoretical fermentation yield in mixotrophic Rubisco-based engineered Escherichia coli

Cited by 38 publications

References 32 publications

Awakening a latent carbon fixation cycle in Escherichia coli

Awakening a latent carbon fixation cycle in Escherichia coli

Synthetic Biology Applied to Carbon Conservative and Carbon Dioxide Recycling Pathways

Awakening a latent carbon fixation cycle inEscherichia coli

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