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Detection of Bile Acids in Complex Matrices Using a Transcription Factor-Based Biosensor.

Kathryn Beabout^1,2, Amy M Ehrenworth Breedon^1,2, Steven M Blum³

¹711th Human Performance Wing, Air Force Research Laboratory, Wright-Patterson Air Force Base, Ohio 45433, United States.

ACS Biomaterials Science & Engineering

|December 8, 2022

Summary

Researchers developed a highly sensitive bile acid sensor using cell-free expression systems. This engineered sensor can detect bile acids in various samples, including wastewater and serum, for health monitoring and environmental safety.

Keywords:

bile acids biosensor cell free protein synthesis deoxycholic acid synthetic biology

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Area of Science:

Biotechnology
Environmental Science
Molecular Biology

Background:

Bile acids are crucial for digestion and human health, present throughout the gastrointestinal tract and excreted in feces.
Their presence in feces makes bile acids potential biomarkers for health monitoring and detecting fecal contamination in water sources.

Purpose of the Study:

To engineer and optimize a sensitive biosensor for detecting bile acids.
To enhance the sensor's utility for real-world applications, including environmental monitoring and health diagnostics.

Main Methods:

Engineered a bile acid sensor in *Escherichia coli* by expressing the transcription factor BreR.
Optimized the sensor through promoter library screening and transitioned expression to a cell-free expression (CFE) system.
Adapted the CFE sensor for detecting bile acids in complex matrices like fecal water, wastewater, and serum, and developed a paper-based assay.

Main Results:

The CFE system significantly increased deoxycholic acid sensitivity by approximately three orders of magnitude compared to the initial engineered sensor.
The optimized sensor demonstrated utility in detecting bile acids across diverse sample types, including environmental and biological fluids.
A paper-based assay format was successfully developed, enhancing the sensor's field applicability.

Conclusions:

The engineered BreR-based biosensor, particularly in its CFE format, offers a highly sensitive and versatile tool for bile acid detection.
This technology holds promise for improved health monitoring and environmental surveillance, enabling early detection of fecal contamination.
The development of a portable paper-based assay further expands the potential for on-site, real-time bile acid analysis.