Self-Assembly of Nanofilaments in Cyanobacteria for Protein Co-localization
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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.
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Synthetic scaffolds and self-assembling protein structures can be used to co-localise sequential pathway enzymes, increase local substrate concentration and improve metabolic channelling, as demonstrated by nanofilament-based protein co-localisation systems in cyanobacteria.
“In addition, synthetic scaffolds and self-assembling protein structures can be used to co-localise sequential pathway enzymes, increase local substrate concentration and improve metabolic channelling, as demonstrated by nanofilament-based protein co-localisation systems in cyanobacteria [169].”
From Cyanobacteria as a Photosynthetic Chassis for Metabolic Pathway Engineering with Heterologous Gene Expression (2026) · cites this paper as [169]
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Experimental — accuracy not yet measuredIn cyanobacteria, the spatial arrangement of photosynthetic membranes, carboxysomes, and central metabolic reactions creates an organised intracellular environment.
“In cyanobacteria, the spatial arrangement of photosynthetic membranes, carboxysomes, and central metabolic reactions creates an organised intracellular environment [169].”
From Cyanobacteria as a Photosynthetic Chassis for Metabolic Pathway Engineering with Heterologous Gene Expression (2026) · cites this paper as [169]
The Results and Discussion section (page unavailable) describes thylakoid membranes aligned along the plasma membrane and carboxysomes aligned in central cytoplasmic regions, supporting an organised intracellular arrangement. The paper does not describe a spatial arrangement of central metabolic reactions as part of that environment. (LLM, conf=0.84)
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References 62 · Cited by 17
- Authors
- Zedler, J. A. Z., Schirmacher, A. M., Russo, D. A., Hodgson, L., Gundersen, E., Matthes, A., Frank, S., Verkade, P., & Jensen, P. E.
- Affiliations
- Friedrich Schiller University Jena (DE)
- University of Bristol (GB)
- University of Copenhagen (DK)
- University College London (GB)
- Corresponding author
- Zedler, Julie A. Z.
- ORCID
- Venue
- ACS Nano
- Publisher
- American Chemical Society (ACS)
- Published
- 2023-12-08
- Volume
- 17
- Issue
- 24
- Pages
- 25279-25290
- ISSN
- 1936-0851 (ISSN-L)
- 1936-086X
- PMC
- PMC10754207
- OpenAlex
- W4389497640
- Cited by
- 17
- References
- 62
- Citations by year
- 2026: 5 · 2025: 7 · 2024: 5
- FWCI
- 1.51
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- ACS Nano (publishedVersion, cc-by-nc-nd)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Research at the University of Copenhagen (University of Copenhagen) (submittedVersion, cc-by-nc-nd)
- UCL Discovery (University College London) (submittedVersion, cc-by-nc-nd)
- Explore Bristol Research (submittedVersion, cc-by)
- Funders
- Novo Nordisk Fonden (NNF19OC0057634)
- European Commission (745959)
- Carlsbergfondet (CF17-0657)
- Alexander von Humboldt-Stiftung
- Research Councils UK (BB/M002969/1)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Structural Characterization
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Keywords
- cyanobacteria, Synechocystis sp. PCC 6803, self-assembly, protein colocalization, heterologous pathway expression, nanobiotechnology
- MeSH terms
- Bacterial Proteins, Photosynthesis, Protein Transport, Synechocystis
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