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.

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Silica gel microspheres are ideal materials for bioencapsulation due to their mechanical properties, biocompatibility, and stability. Encapsulated cells are isolated from the environment and protected from predators, changes in pH, and osmotic stress. However methods for the production of silica gel microspheres suitable for bioencapsulation are not well established. This paper describes a method for the production of monodisperse silicon alkoxide cross-linked silica nanoparticle (SNP) gel microspheres for bioencapsulation in which silica gel precursor is extruded from a needle into a cross-flowing stream of mineral oil. Microspheres produced ranged from 1.3 to 2.9 mm in diameter with coefficients of variation ranging from 2 to 6%. Microsphere size was mainly controlled by the flowrate of the cross-flowing oil and smaller microspheres generally had larger coefficients of variation. The method described in this paper can be optimised to produce silica gel microspheres with a diverse range of compositions and properties.

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 was engineered to display SpyTags on polar flagella for cell attachment and cross-linking of SpyCatcher modified secreted scaffolds. Through deletion of the autolysis LytC, endospore limited B. subtilis cells become a structural component of the material with spores for long-term storage of genetic programming. Known silica biomineralization peptides were 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 silica material and that new functions can be readily 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 as functional, self-healing materials for use as responsive coatings and plasters.