Abstract
Engineered living materials (ELMs) are a fast-growing area of research that combine approaches in synthetic biology and material science. Here, we engineer B. subtilis to become a living component of a silica material composed of self-assembling protein scaffolds for functionalization and cross-linking of cells. B. subtilis is engineered to display SpyTags on polar flagella for cell attachment to SpyCatcher modified secreted scaffolds. We engineer endospore limited B. subtilis cells to become a structural component of the material with spores for long-term storage of genetic programming. Silica biomineralization peptides are screened and scaffolds designed for silica polymerization to fabricate biocomposite materials with enhanced mechanical properties. 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. subtilis strains. We believe that this work will serve as a framework for the future design of resilient ELMs.
Highlights
Engineered living materials (ELMs) are a fast-growing area of research that combine approaches in synthetic biology and material science
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
We show that B. subtilis can be engineered to display SpyTags on clusters of polar flagella for cell attachment and cross-linking of EutM scaffolds that constitute the protein matrix of our ELM
Summary
Engineered living materials (ELMs) are a fast-growing area of research that combine approaches in synthetic biology and material science. We engineer B. subtilis to become a living component of a silica material composed of self-assembling protein scaffolds for functionalization and cross-linking of cells. 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] 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. We sought to broaden the ELM landscape by engineering a resilient ELM biocomposite that uses the sporeforming bacteria Bacillus subtilis as its living component for the secretion of self-assembling protein scaffolds for cell cross-linking and silica biomineralization. We believe that this work will serve as a framework for the future design of resilient ELMs as functional, self-healing materials for use as coatings and plasters that can respond to external stimuli due to the functions provided by the engineered cells in such materials
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