Sun-Young Kang^1,2,4, Anaya Pokhrel^1,2,4, Sara Bratsch^1,2,4, Joey J. Benson³, Seung-Oh Seo^1,2, Maureen B. Quin^1,2, Alptekin Aksan^2,3 & Claudia Schmidt-Dannert^1,2,*

¹Department of Biochemistry, Molecular Biology & Biochemistry, University of Minnesota, Minneapolis, MN 55455, USA. ²BioTechnology Institute, University of Minnesota, St. Paul, MN 55108, USA. ³Department of Mechanical Engineering, University of Minnesota, Minneapolis, MN 55455, USA. ⁴These authors contributed equally: Sun-Young Kang, Anaya Pokhrel, Sara Bratsch.

^*Corresponding author.

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.