Using Light-Activated Enzymes for Modulating Intracellular c-di-GMP Levels in Bacteria

Area of Science:

• Bacterial Physiology
• Microbial Signaling
• Synthetic Biology

Background:

• The second messenger cyclic di-GMP (c-di-GMP) is crucial for regulating diverse bacterial behaviors.
• Understanding and controlling c-di-GMP levels are key to deciphering bacterial physiology.

Purpose of the Study:

• To develop light-inducible tools for precise manipulation of intracellular c-di-GMP concentrations.
• To demonstrate the application of these tools in controlling bacterial phenotypes.

Main Methods:

• Engineered a red light-activated diguanylate cyclase (synthesizes c-di-GMP).
• Engineered a blue light-activated phosphodiesterase (degrades c-di-GMP).
• Applied these enzymes to manipulate c-di-GMP levels in *E. coli* and *Azospirillum brasilense*.

Main Results:

• Demonstrated light-dependent regulation of bacterial motility and biofilm formation in *E. coli*.
• Showcased light-controlled chemotactic behavior in *Azospirillum brasilense*.
• Validated the enzymes' efficacy in modulating c-di-GMP dependent processes.

Conclusions:

• Light-activated enzymes provide a powerful method for spatiotemporal control of bacterial c-di-GMP signaling.
• These tools enable precise regulation of bacterial phenotypes like motility and biofilm formation.
• The developed system holds potential for studying fast biological processes and single-cell bacterial behavior.