Kinase FRET biosensors for rapid fluorescent detection of acrylamide

Overview

This solution is a biosensing platform that uses fluorescence resonance energy transfer (FRET) in yeast cells to detect acrylamide, a known contaminant of concern in drinking water and certain cooked foods. The approach combines phosphoproteomic screening with engineered kinase-based FRET biosensors to produce a rapid, quantitative fluorescent readout when acrylamide is present in a water sample. The goal is a low-limit, dose-dependent detection method that responds within seconds to minutes, enabling faster and more accessible acrylamide monitoring compared with conventional laboratory techniques.

Technical specifications

• FRET-based detection in yeast: Yeast cells are genetically modified to express FRET biosensors whose sensory domain contains phosphorylation substrates specific to acrylamide-responsive kinases. When acrylamide is present, kinase activity changes phosphorylation state, altering FRET efficiency and producing a measurable fluorescent signal.
• Phosphoproteomics-guided design: Quantitative phosphoproteomic analysis of yeast cells exposed to acrylamide is used to identify the kinases most sensitive to acrylamide and their downstream substrates, improving biosensor specificity and sensitivity.
• Rapid response: Because kinase-driven phosphorylation events occur on timescales of seconds to minutes, the biosensor is designed to generate near real-time fluorescent responses.
• Quantitative performance targets: Dose-response curves, detection limit, and response time will be evaluated to characterize sensitivity and dynamic range.
• Application areas: Water quality monitoring, food safety testing for acrylamide in processed foods, and broader environmental contaminant screening where kinase-pathway disruption can serve as a sensitive indicator.

Technology readiness level

The technology is at an early proof-of-concept stage. Preliminary data from the co-PI has demonstrated that in vivo protein phosphorylation by kinases responds to changes in the extracellular environment, supporting the biological basis for the biosensor design. The proposed 6-month project will screen acrylamide-sensitive kinases via phosphoproteomics, engineer yeast strains expressing kinase-specific FRET biosensors, and evaluate dose-response, detection limit, and response time. The deliverable is a proof-of-concept demonstration of phosphoproteomics-guided FRET biosensor design for acrylamide detection, with further optimization and validation needed before commercial deployment.