Engineering the S-Layer of Caulobacter crescentus as a Foundation for Stable, High-Density, 2D Living Materials
| Autor: | Paul D. Ashby, Sneha Jani, Kathleen R. Ryan, Behzad Rad, Dong Li, Bruce E. Cohen, Victor R. Mann, Marimikel Charrier, Caroline M. Ajo-Franklin, Lisa Yun |
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| Rok vydání: | 2018 |
| Předmět: |
DNA
Bacterial engineered living materials 0106 biological sciences Materials science Caulobacter Biomedical Engineering quantum dots Bioengineering Nanotechnology Paracrystalline Cyanobacteria Regenerative Medicine 01 natural sciences Biochemistry Genetics and Molecular Biology (miscellaneous) Article law.invention Protein filament Medicinal and Biomolecular Chemistry 03 medical and health sciences Synthetic biology RsaA Underpinning research Confocal microscopy law 010608 biotechnology Caulobacter crescentus 030304 developmental biology Gene Editing 0303 health sciences biology Cell Membrane Bacterial biomaterial Biomaterial DNA General Medicine biology.organism_classification 1.3 Chemical and physical sciences Biochemistry and Cell Biology S-layer |
| Zdroj: | ACS synthetic biology, vol 8, iss 1 Charrier, Marimikel; Li, Dong; Mann, Victor R; Yun, Lisa; Jani, Sneha; Rad, Behzad; et al.(2019). Engineering the S-Layer of Caulobacter crescentus as a Foundation for Stable, High-Density, 2D Living Materials. ACS SYNTHETIC BIOLOGY, 8(1), 181-190. doi: 10.1021/acssynbio.8b00448. Lawrence Berkeley National Laboratory: Retrieved from: http://www.escholarship.org/uc/item/7bs7v7pm |
| ISSN: | 2161-5063 |
| DOI: | 10.1021/acssynbio.8b00448 |
| Popis: | Materials synthesized by organisms, such as bones and wood, combine the ability to self-repair with remarkable mechanical properties. This multifunctionality arises from the presence of living cells within the material and hierarchical assembly of different components across nanometer to micron scales. While creating engineered analogues of these natural materials is of growing interest, our ability to hierarchically order materials using living cells largely relies on engineered 1D protein filaments. Here, we lay the foundation for bottom-up assembly of engineered living material composites in 2D along the cell body using a synthetic biology approach. We engineer the paracrystalline surface-layer (S-layer) of Caulobacter crescentus to display SpyTag peptides that form irreversible isopeptide bonds to SpyCatcher-modified proteins, nanocrystals, and biopolymers on the extracellular surface. Using flow cytometry and confocal microscopy, we show that attachment of these materials to the cell surface is uniform, specific, and covalent, and its density can be controlled on the basis of the insertionlocationwithin the S-layer protein, RsaA. Moreover, we leverage the irreversible nature of this attachment to demonstrate via SDS-PAGE that the engineered S-layer can display a high density of materials, reaching 1 attachment site per 288 nm2. Finally, we show that ligation of quantum dots to the cell surface does not impair cell viability, and this composite material remains intact over a period of 2 weeks. Taken together, this work provides a platform for self-organization of soft and hard nanomaterials on a cell surface with precise control over 2D density, composition, and stability of the resulting composite, and is a key step toward building hierarchically ordered engineered living materials with emergent properties. |
| Databáze: | OpenAIRE |
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