De novo Autogenic Engineered Living Functional Materials

Autogenic engineered living materials (ELMs) involve in situ production and engineering of native extracellular matrix (ECM). However, the existing autogenic ELMs have limited scope and functionalities. Herein, we report a platform for de novo autogenic functional ELMs. By protein mining, we have discovered CsgA-like 33,564 homologs that can find potential utility as de novo ECM of protein nanofibers. By employing AlphaFold2 and molecular dynamics simulations, we shed insights into the CsgA-like β-solenoid protein structures and stability. By hacking Escherichia coli curli machinery, we demonstrate the production of de novo autogenic ELMs from CsgA-like proteins (≤9-times the molecular weight and β-sheet repeat units) of extremophilic non-model bacteria. Additionally, we biomanufacture macroscopic materials with tunable mechanical properties (enhancing storage modulus by 3-times) and programmable functionalities (3D printability, binding to nanoparticles/antibodies). This work showcases a versatile platform to discover, rationally design, and harness the sophisticated functionalities of natural systems for futuristic autogenic ELMs.