Bacterial Cell Display as a Robust and Versatile Platform for Engineering Low-Affinity Ligands and Enzymes

Area of Science:

• Synthetic Biology
• Protein Engineering
• Biotechnology

Background:

• Directed evolution expands biological capabilities but relies on robust selection platforms.
• Bacterial cell display is effective for enzyme engineering but adoption is limited.
• Developing new selection platforms is costly and time-consuming.

Purpose of the Study:

• To develop a quantitative reporter (SNAP) for bacterial cell display.
• To improve the robustness and flexibility of bacterial cell display platforms.
• To enable efficient engineering of enzymes and DNA-binding proteins.

Main Methods:

• Development of SNAP as a quantitative reporter for bacterial cell display.
• Demonstration of SNAP's utility in troubleshooting and platform development.
• Application of bacterial cell display for functional characterization of DNA-binding proteins and enzymes.

Main Results:

• SNAP enables fast troubleshooting and systematic development of bacterial cell display.
• The platform successfully harnesses weak protein-nucleic acid interactions for cell labeling.
• Bacterial cell display proved robust and flexible for engineering ligands and enzymes.

Conclusions:

• Bacterial cell display, enhanced by SNAP, is a robust and flexible platform.
• This system facilitates the systematic engineering of proteins for XNA molecular biology.
• The developed platform accelerates the expansion of biological functions through directed evolution.