Accelerating the design of pili-enabled living materials using an integrative technological workflow

Area of Science:

• Synthetic Biology
• Biomaterials Engineering
• Bioinformatics

Background:

• Engineered Living Materials (ELMs) offer self-healing and evolvable functionalities.
• Development of ELMs is limited by the lack of nonpathogenic bacterial chassis and programmable biopolymers.
• Need for advanced tools to design and produce novel biopolymers for ELMs.

Purpose of the Study:

• To establish a technological workflow for designing ELMs by integrating bioinformatics, structural biology, and synthetic biology.
• To develop a bioinformatics tool for identifying novel biopolymers and producing bacteria.
• To engineer programmable pili for ELM applications and demonstrate biomass conversion.

Main Methods:

• Development of Bacteria Biopolymer Sniffer (BBSniffer) software for biopolymer mining.
• Identification and characterization of the covalently linked pili (CLP) gene cluster in Corynebacterium glutamicum.
• Genetic manipulation, structural biology, and synthetic biology approaches for CLP assembly and ELM engineering.

Main Results:

• BBSniffer successfully identified the CLP biosynthetic gene cluster from a pathogenic pilus.
• The molecular mechanism of CLP assembly was elucidated, enabling programmable pili.
• Engineered ELMs demonstrated the conversion of cellulosic biomass into lycopene.

Conclusions:

• A novel technological workflow facilitates the rational design of ELMs.
• Programmable pili derived from engineered bacteria represent a new platform for ELM development.
• This approach enables the creation of functional living materials with bioconversion capabilities.