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An efficient design strategy for a whole-cell biosensor based on engineered ribosome binding sequences.

Qing Yu¹, Yan Li, Anzhou Ma

¹Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing, China.

Analytical and Bioanalytical Chemistry

|September 28, 2011

Summary

Engineered ribosome binding sequences (RBS) significantly enhance whole-cell biosensor performance. Optimized RBS boosted signal intensity, lowered detection limits, and accelerated response times for BTEX detection.

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Area of Science:

Microbiology
Molecular Biology
Environmental Science

Background:

Ribosome binding sequences (RBS) in prokaryotes are crucial for mRNA translation and stability.
Whole-cell biosensors are valuable tools for environmental monitoring.
Optimizing RBS is a potential strategy to improve biosensor efficacy.

Purpose of the Study:

To investigate the impact of engineered ribosome binding sequences (RBS) on the sensitivity and signal intensity of environmental whole-cell biosensors.
To compare the performance of biosensors with different RBS designs for detecting benzene, toluene, ethylbenzene, and xylenes (BTEX).

Main Methods:

Construction of three Escherichia coli-based whole-cell biosensors with identical Pu promoter and xylR regulator but distinct engineered RBS.
Evaluation of biosensor performance using time- and dose-dependent luminescence induction assays with 2-chlorotoluene.
Analysis of signal intensity, limit of detection (LOD), and response speed.

Main Results:

Engineered RBS (BTEX-SE and BTEX-SD) resulted in 10-35 times higher luminescence signal intensity compared to the standard RBS (BTEX-W).
The LOD for BTEX-SE and BTEX-SD biosensors was significantly lower (20-25 μmol L(-1)) than for BTEX-W (120 μmol L(-1)).
BTEX-SE and BTEX-SD biosensors exhibited a three-fold faster response time to analytes.

Conclusions:

Rationally designed RBS in the 5' untranslated region (5' UTR) of reporter genes can substantially enhance whole-cell biosensor performance.
Engineered RBS offer a promising strategy for improving sensitivity, signal intensity, and response speed in environmental biosensing applications.
This approach holds potential for more effective detection of BTEX compounds and other environmental pollutants.