Incorporation of Functional Rubisco Activases into Engineered Carboxysomes to Enhance Carbon Fixation.

Chen, Taiyu, Fang, Yi, Jiang, Qiuyao, Dykes, Gregory F, Lin, Yongjun ORCID: 0000-0002-3818-4816, Price, G Dean, Long, Benedict M ORCID: 0000-0002-4616-2967 and Liu, Lu-Ning
(2021) Incorporation of Functional Rubisco Activases into Engineered Carboxysomes to Enhance Carbon Fixation. ACS synthetic biology.

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The carboxysome is a versatile paradigm of prokaryotic organelles and is a proteinaceous self-assembling microcompartment that plays essential roles in carbon fixation in all cyanobacteria and some chemoautotrophs. The carboxysome encapsulates the central CO<sub>2</sub>-fixing enzyme, ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), using a polyhedral protein shell that is selectively permeable to specific metabolites in favor of Rubisco carboxylation. There is tremendous interest in repurposing carboxysomes to boost carbon fixation in heterologous organisms. Here, we develop the design and engineering of α-carboxysomes by coexpressing the Rubisco activase components CbbQ and CbbO with α-carboxysomes in <i>Escherichia coli</i>. Our results show that CbbQ and CbbO could assemble into the reconstituted α-carboxysome as intrinsic components. Incorporation of both CbbQ and CbbO within the carboxysome promotes activation of Rubisco and enhances the CO<sub>2</sub>-fixation activities of recombinant carboxysomes. We also show that the structural composition of these carboxysomes could be modified in different expression systems, representing the plasticity of the carboxysome architecture. In translational terms, our study informs strategies for engineering and modulating carboxysomes in diverse biotechnological applications.

Item Type: Article
Divisions: Faculty of Health and Life Sciences
Faculty of Health and Life Sciences > Institute of Systems, Molecular and Integrative Biology
Depositing User: Symplectic Admin
Date Deposited: 20 Oct 2021 08:42
Last Modified: 18 Jan 2023 21:26
DOI: 10.1021/acssynbio.1c00311

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