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Self-Assembly of Nanofilaments in Cyanobacteria for Protein Co-localization

Zedler, Julie AZ; Schirmacher, Alexandra M; Russo, David A; Hodgson, Lorna; Gundersen, Emil; Matthes, Annemarie; Frank, Stefanie; ... Jensen, Poul Erik; + view all (2023) Self-Assembly of Nanofilaments in Cyanobacteria for Protein Co-localization. ACS Nano 10.1021/acsnano.3c08600. (In press).

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Abstract

Cyanobacteria offer great potential as alternative biotechnological hosts due to their photoautotrophic capacities. However, in comparison to established heterotrophic hosts, several key aspects, such as product titers, are still lagging behind. Nanobiotechnology is an emerging field with great potential to improve existing hosts, but so far, it has barely been explored in microbial photosynthetic systems. Here, we report the establishment of large proteinaceous nanofilaments in the unicellular model cyanobacterium Synechocystis sp. PCC 6803 and the fast-growing cyanobacterial strain Synechococcus elongatus UTEX 2973. Transmission electron microscopy and electron tomography demonstrated that expression of pduA*, encoding a modified bacterial microcompartment shell protein, led to the generation of bundles of longitudinally aligned nanofilaments in S. elongatus UTEX 2973 and shorter filamentous structures in Synechocystis sp. PCC 6803. Comparative proteomics showed that PduA* was at least 50 times more abundant than the second most abundant protein in the cell and that nanofilament assembly had only a minor impact on cellular metabolism. Finally, as a proof-of-concept for co-localization with the filaments, we targeted a fluorescent reporter protein, mCitrine, to PduA* by fusion with an encapsulation peptide that natively interacts with PduA. The establishment of nanofilaments in cyanobacterial cells is an important step toward cellular organization of heterologous pathways and the establishment of cyanobacteria as next-generation hosts.

Type: Article
Title: Self-Assembly of Nanofilaments in Cyanobacteria for Protein Co-localization
Location: United States
DOI: 10.1021/acsnano.3c08600
Publisher version: https://doi.org/10.1021/acsnano.3c08600
Language: English
Additional information: This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions.
Keywords: Bacteria, Carbohydrates, Chemical structure, Fluorescence, Peptides and proteins
UCL classification: UCL
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering
URI: https://discovery.ucl.ac.uk/id/eprint/10184352
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