[Characterization and application of several lysis cassettes]

Area of Science:

• Synthetic biology
• Microbiology

Context:

• Lysis modules are crucial in synthetic biology for genetic circuit design.
• Phage-derived lysis cassettes offer a method for controlled bacterial lysis.
• Detailed characterization of these lysis cassettes is lacking.

Purpose:

• To characterize the functional behavior of five distinct lysis cassettes in *Escherichia coli* Top10 and *Pseudomonas aeruginosa* PAO1.
• To evaluate the efficacy of arabinose- and rhamnose-inducible systems for controlling lysis cassette expression.
• To engineer a novel strain for light-induced surface modification via controlled lysis.

Summary:

• Five lysis cassettes (S, A52G, C51S S76C, LKD, LUZ) were expressed inducibly in *E. coli* Top10, showing varied lysis behaviors based on growth stage, inducer concentration, and plasmid copy number.
• Challenges in constructing inducible lysis systems in *P. aeruginosa* PAO1 due to background expression were overcome by chromosomal integration of a rhamnose-inducible lysis cassette, with LUZ and LKD proving most effective.
• An engineered strain (Q16) combining an optogenetic module (BphS) and the LUZ lysis cassette achieved light-induced lysis and surface adhesion, demonstrating potential for surface modification.

Impact:

• Provides a comprehensive characterization of bacterial lysis cassettes, aiding synthetic biology applications.
• Identifies effective lysis cassettes and inducible systems for both *E. coli* and *P. aeruginosa*.
• Demonstrates a novel platform for light-controlled bacterial lysis and surface modification, with potential in biotechnology and materials science.