Engineering Green-light-responsive Heterologous Gene Expression in Pseudomonas.

Area of Science:

• Synthetic Biology
• Microbial Engineering
• Optogenetics

Background:

• Precise control of bacterial gene expression is crucial for engineered applications.
• Chemical inducers can leave residual compounds, complicating biological systems.
• Light-inducible systems offer a cleaner alternative for controlling cellular functions.

Purpose of the Study:

• To engineer biofilm formation in Pseudomonas putida using optogenetics.
• To adapt the cyanobacterial CcaSR two-component system for light-controlled gene expression.
• To provide a method for precise ON/OFF switching of bacterial functions without chemical additives.

Main Methods:

• Utilized the CcaSR two-component system for light-mediated transcriptional control.
• Placed the expression of diguanylate cyclase PleD under CcaSR system regulation.
• Applied optogenetic control to induce biofilm formation in Pseudomonas putida.

Main Results:

• Successfully induced biofilm formation using light as the sole trigger.
• Demonstrated precise ON/OFF control of gene expression via the CcaSR system.
• Achieved regulation without the need for chemical inducers.

Conclusions:

• Optogenetic control via the CcaSR system is effective for inducing biofilm formation.
• This light-based method provides a cleaner and more cost-effective approach compared to chemical induction.
• Enables enhanced exploitation of beneficial biofilm features through precise genetic regulation.