![]() In addition, because spores contain the genetic information that was programmed into engineered vegetative cells, living materials may be autonomously fabricated at the sites of use from stored spores. This was recently demonstrated by bioprinting of Bacillus spores in an agarose matrix 6. subtilis cells will therefore be able to enter a dormant spore state in our ELM under unfavorable environmental conditions, allowing it to persist until favorable conditions induce germination and cell revival. Importantly, it forms spores that remain viable for a long time and allow the bacteria to survive extreme conditions 29, 30, 31. subtilis is an industrially used GRAS (Generally Recognized as Safe) bacteria known to have excellent protein secretion capabilities. Here, we sought to broaden the ELM landscape by engineering a resilient ELM biocomposite that uses the spore-forming bacteria Bacillus subtilis as its living component for the secretion of self-assembling protein scaffolds for cell cross-linking and silica biomineralization.ī. In addition, ELM fabrication is currently limited to common chassis organisms that lack the resilience and long-term viability capabilities to withstand conditions outside of the laboratory. Among these examples, the secretion of a polypeptide-based scaffolding system or matrix offers greater control over material assembly and functionalization due to the genetic programmability of polypeptide structures and functions.ĭespite the advances in ELM design, the diversity of different ELM types capable of autonomous self-fabrication and regeneration is still small. Other types of extracellular matrices for ELM fabrication were created from secreted bacterial cellulose to embed microbial cells 26, 27 or from elastin-like polypeptides to attach Caulobacter cells via their protein S-layers 28. True ELMs have been created by engineering Escherichia coli to produce an extracellular matrix from curli fibers 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25. Engineering of ‘truly’ living materials where the living component actively facilitates material fabrication and organization is much more challenging. The fabrication of most ELMs, however, has so far largely relied on physical methods for incorporating a living component in an external material 4, 5, 6, 7. Engineered living materials (ELMs) are therefore a new and fast-growing area of research that combines approaches in synthetic biology and material sciences 1, 2, 3, 4. The design of cells capable of producing self-organizing, biocomposite materials has the potential to enable the fabrication of new types of functional living materials with the ability of self-fabrication and self-repair. We believe that this work will serve as a framework for the future design of resilient ELMs. We show that the resulting ELM can be regenerated from a piece of cell containing silica material and that new functions can be incorporated by co-cultivation of engineered B. Silica biomineralization peptides are screened and scaffolds designed for silica polymerization to fabricate biocomposite materials with enhanced mechanical properties. subtilis cells to become a structural component of the material with spores for long-term storage of genetic programming. subtilis is engineered to display SpyTags on polar flagella for cell attachment to Sp圜atcher modified secreted scaffolds. subtilis to become a living component of a silica material composed of self-assembling protein scaffolds for functionalization and cross-linking of cells. Engineered living materials (ELMs) are a fast-growing area of research that combine approaches in synthetic biology and material science. ![]()
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