Carboxysome encapsulation of the CO2-fixing enzyme Rubisco in tobacco chloroplasts

Benedict M. Long (), Wei Yih Hee, Robert E. Sharwood, Benjamin D. Rae, Sarah Kaines, Yi-Leen Lim, Nghiem D. Nguyen, Baxter Massey, Soumi Bala, Susanne von Caemmerer, Murray R. Badger and G. Dean Price
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Benedict M. Long: The Australian National University
Wei Yih Hee: The Australian National University
Robert E. Sharwood: The Australian National University
Benjamin D. Rae: The Australian National University
Sarah Kaines: The Australian National University
Yi-Leen Lim: The Australian National University
Nghiem D. Nguyen: The Australian National University
Baxter Massey: The Australian National University
Soumi Bala: The Australian National University
Susanne von Caemmerer: The Australian National University
Murray R. Badger: The Australian National University
G. Dean Price: The Australian National University

Nature Communications, 2018, vol. 9, issue 1, 1-14

Abstract: Abstract A long-term strategy to enhance global crop photosynthesis and yield involves the introduction of cyanobacterial CO2-concentrating mechanisms (CCMs) into plant chloroplasts. Cyanobacterial CCMs enable relatively rapid CO2 fixation by elevating intracellular inorganic carbon as bicarbonate, then concentrating it as CO2 around the enzyme Rubisco in specialized protein micro-compartments called carboxysomes. To date, chloroplastic expression of carboxysomes has been elusive, requiring coordinated expression of almost a dozen proteins. Here we successfully produce simplified carboxysomes, isometric with those of the source organism Cyanobium, within tobacco chloroplasts. We replace the endogenous Rubisco large subunit gene with cyanobacterial Form-1A Rubisco large and small subunit genes, along with genes for two key α-carboxysome structural proteins. This minimal gene set produces carboxysomes, which encapsulate the introduced Rubisco and enable autotrophic growth at elevated CO2. This result demonstrates the formation of α-carboxysomes from a reduced gene set, informing the step-wise construction of fully functional α-carboxysomes in chloroplasts.

Date: 2018
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DOI: 10.1038/s41467-018-06044-0

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