Enhancing DNT Detection by a Bacterial Bioreporter: Directed Evolution of the Transcriptional Activator YhaJ

Overview

Journal Front Bioeng Biotechnol

Date 2022 Mar 3

PMID 35237579

Authors

Tal Elad

Benjamin Shemer

Shilat Simanowitz

Yossef Kabessa

Yosef Mizrachi

Azriel Gold

Etai Shpigel

Aharon J Agranat

Shimshon Belkin

Affiliations

Soon will be listed here.

Abstract

Detection of buried landmines is a dangerous and complicated task that consumes large financial resources and poses significant risks to the personnel involved. A potential alternative to conventional detection methodologies is the use of microbial bioreporters, capable of emitting an optical signal upon exposure to explosives, thus revealing to a remote detector the location of buried explosive devices. We have previously reported the design, construction, and optimization of an -based bioreporter for the detection of 2,4,6-trinitrotoluene (TNT) and its accompanying impurity 2,4-dinitrotoluene (DNT). Here we describe the further enhancement of this bioreporter by the directed evolution of YhaJ, the transcriptional activator of the gene promoter, the sensing element of the bioreporter's molecular circuit. This process resulted in a 37-fold reduction of the detection threshold, as well as significant enhancements to signal intensity and response time, rendering this sensor strain more suitable for detecting the minute concentrations of DNT in the soil above buried landmines. The capability of this enhanced bioreporter to detect DNT buried in sand is demonstrated.

Citing Articles

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High-affinity promotor binding of YhaJ mediates a low signal leakage for effective DNT detection.

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Structural basis of transcription factor YhaJ for DNT detection.

Kim M, Kang R, Jeon T, Ryu S iScience. 2023; 26(10):107984.

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