Protein Engineering of a Genetically Encodable Biosensor for Wastewater Detection of Profen NSAIDs

Area of Science:

• Environmental Science
• Biotechnology
• Chemical Engineering

Background:

• Non-steroidal anti-inflammatory drugs (NSAIDs) are widespread environmental contaminants.
• The "profen" class of NSAIDs poses risks to aquatic ecosystems due to incomplete wastewater treatment.
• Current monitoring methods lack high-frequency testing capabilities at point sources.

Purpose of the Study:

• To engineer a genetically encodable, protein-based biosensor for detecting specific NSAIDs in wastewater effluent.
• To develop a scalable and cost-effective alternative for real-time pharmaceutical pollution monitoring.

Main Methods:

• Computational protein design, deep mutational scanning, and yeast screening were used to repurpose the PYR1 receptor.
• The engineered sensor, PYR1^NSAID, was tested for its detection limits and specificity.
• A split Nanoluc luminescence assay was employed to demonstrate dose-responsive activity in simulated wastewater.

Main Results:

• The PYR1^NSAID biosensor exhibits nanomolar detection limits for ketoprofen and pranoprofen.
• It shows µM sensitivity to other NSAIDs including ibuprofen, fenoprofen, tolmetin, and diclofenac.
• The sensor demonstrated dose-responsive activity in complex wastewater matrices.

Conclusions:

• PYR1^NSAID represents a significant advancement in biosensing technology for pharmaceutical pollutants.
• This protein-based sensor offers a promising foundation for real-time monitoring of NSAIDs in environmental samples.
• The development paves the way for more effective management of pharmaceutical pollution in aquatic ecosystems.