De Novo Autogenic Engineered Living Functional Materials

Area of Science:

• Synthetic Biology
• Biomaterials Science
• Protein Engineering

Background:

• Autogenic engineered living materials (ELMs) facilitate in situ production of native extracellular matrix (ECM).
• Current autogenic ELMs have limitations in scope and functionality.
• A need exists for versatile platforms to create advanced autogenic ELMs.

Purpose of the Study:

• To present a versatile platform for de novo autogenic functional ELMs.
• To leverage protein mining, computational modeling, and synthetic biology for ELM development.
• To engineer novel biomaterials with enhanced mechanical and protective properties.

Main Methods:

• Analyzed 33,564 CsgA-like homologs to identify ECM protein nanofiber candidates.
• Utilized AlphaFold2 and molecular dynamics simulations to assess structural stability of β-solenoid proteins.
• Reprogrammed Escherichia coli curli machinery for CsgA-like ELM biosynthesis in non-model bacteria.

Main Results:

• Identified and engineered novel CsgA-like ELMs with up to 9-fold increased molecular weight and expanded β-sheet repeats.
• Fabricated macroscopic biomaterials with a 3-fold increase in storage modulus.
• Demonstrated UV-C attenuation, extending embedded cell survival by 5-fold, and achieved 3D printability and selective binding.

Conclusions:

• Established a powerful framework for discovering and designing autogenic ELMs.
• Advanced the field of engineered living materials with enhanced functionality and programmability.
• Paved the way for next-generation biomaterials harnessing natural biomolecular systems.