Engineering carbon fixation in E. coli : from heterologous RuBisCO expression to the Calvin–Benson–Bassham cycle

Antonovsky, Niv; Gleizer, Shmuel; Milo, Ron

doi:10.1016/j.copbio.2017.06.006

Cited by 45 publications

(32 citation statements)

References 62 publications

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“…Together with an appropriate energy supply, introducing RuBisCO and PRK would allow heterotrophic microorganisms to x CO 2 into carbohydrates. 12 Re-xing the CO 2 released during the carbon metabolism is a promising strategy to increase the carbon usage efficiency in heterotrophic microorganisms. Therefore, studying the CBB cycle and increasing its efficiency will potentially have benecial effects on the efficiencies of both autotrophic and heterotrophic cell factories.…”

Section: The Calvin-benson-bassham (Cbb) Cyclementioning

confidence: 99%

Engineering photoautotrophic carbon fixation for enhanced growth and productivity

Liang¹,

Lindberg²,

Lindblad³

2018

Sustainable Energy Fuels

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Section: The Calvin-benson-bassham (Cbb) Cyclementioning

confidence: 99%

Engineering photoautotrophic carbon fixation for enhanced growth and productivity

Liang¹,

Lindberg²,

Lindblad³

2018

Sustainable Energy Fuels

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“…To avoid the constraints a photosynthetic environment may pose on the catalytic evolution of Rubisco, modern laboratory evolution applications have made particular use of Rubisco-dependent E. coli (RDE) screens (12,21,22). An elegant advance has been the extensive rewiring of E. coli metabolism to incorporate a non-native CBB cycle where cell survival can be made dependent on CO 2 -fixation by Rubisco (23).…”

mentioning

confidence: 99%

An improved Escherichia coli screen for Rubisco identifies a protein–protein interface that can enhance CO2-fixation kinetics

Wilson

Avila

Conlan³

et al. 2018

Journal of Biological Chemistry

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An overarching goal of photosynthesis research is to identify how components of the process can be improved to benefit crop productivity, global food security, and renewable energy storage. Improving carbon fixation has mostly focused on enhancing the CO fixing enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). This grand challenge has mostly proved ineffective because of catalytic mechanism constraints and required chaperone complementarity that hinder Rubisco biogenesis in alternative hosts. Here we refashion metabolism by expressing a phosphoribulokinase-neomycin phosphotransferase fusion protein to produce a high-fidelity, high-throughput Rubisco-directed evolution (RDE2) screen that negates false-positive selection. Successive evolution rounds using the plant-like-Rubisco from the cyanobacterium BP1 identified two large subunit and six small subunit mutations that improved carboxylation rate, efficiency, and specificity. Structural analysis revealed the amino acids clustered in an unexplored subunit interface of the holoenzyme. To study its effect on plant growth, the-Rubisco was transformed into tobacco by chloroplast transformation. As previously seen for PCC6301 Rubisco, the specialized folding and assembly requirements of-Rubisco hinder its heterologous expression in leaf chloroplasts. Our findings suggest that the ongoing efforts to improve crop photosynthesis by integrating components of a cyanobacteria CO-concentrating mechanism will necessitate co-introduction of the ancillary molecular components required for Rubisco biogenesis.

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“…Synthetic photosynthesis has made exciting progress recently via the incorporation of non-native Calvin-Benson-like carbon assimilation in Escherichia coli . The highlight was the production of a bacterium that is capable of making sugars and other life-preserving metabolites from atmospheric CO 2 88 , 89 . Although at this stage both energy and reducing power are required through the oxidation of an external organic acid in an isolated metabolic module and thus no net carbon gain is achieved, this discovery clearly proves the potential of synthetic biology to optimize pathways of biotechnological significance and may even lead to new avenues for optimizing CO 2 fixation in plants 89 .…”

Section: Synthetic Biology For Co 2 Fixation In Nomentioning

confidence: 99%

“…The highlight was the production of a bacterium that is capable of making sugars and other life-preserving metabolites from atmospheric CO 2 88 , 89 . Although at this stage both energy and reducing power are required through the oxidation of an external organic acid in an isolated metabolic module and thus no net carbon gain is achieved, this discovery clearly proves the potential of synthetic biology to optimize pathways of biotechnological significance and may even lead to new avenues for optimizing CO 2 fixation in plants 89 . Along such lines is the successful development of a synthetic carbon fixation pathway that functions efficiently in vitro but faces significant challenges for it to be compatible in a biological context 90 .…”

Section: Synthetic Biology For Co 2 Fixation In Nomentioning

confidence: 99%

Engineering photosynthesis: progress and perspectives

Orr

Pereira

et al. 2017

F1000Res

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Photosynthesis is the basis of primary productivity on the planet. Crop breeding has sustained steady improvements in yield to keep pace with population growth increases. Yet these advances have not resulted from improving the photosynthetic process per se but rather of altering the way carbon is partitioned within the plant. Mounting evidence suggests that the rate at which crop yields can be boosted by traditional plant breeding approaches is wavering, and they may reach a “yield ceiling” in the foreseeable future. Further increases in yield will likely depend on the targeted manipulation of plant metabolism. Improving photosynthesis poses one such route, with simulations indicating it could have a significant transformative influence on enhancing crop productivity. Here, we summarize recent advances of alternative approaches for the manipulation and enhancement of photosynthesis and their possible application for crop improvement.

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Engineering carbon fixation in E. coli : from heterologous RuBisCO expression to the Calvin–Benson–Bassham cycle

Cited by 45 publications

References 62 publications

Engineering photoautotrophic carbon fixation for enhanced growth and productivity

Engineering photoautotrophic carbon fixation for enhanced growth and productivity

An improved Escherichia coli screen for Rubisco identifies a protein–protein interface that can enhance CO2-fixation kinetics

Engineering photosynthesis: progress and perspectives

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