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Tracing explosives in soil with transcriptional regulators of Pseudomonas putida evolved for responding to nitrotoluenes
Although different biological approaches for detection of anti‐personnel mines and other unexploded ordnance (UXO) have been entertained, none of them has been rigorously documented thus far in the scientific literature. The industrial 2,4,6 trinitrotoluene (TNT) habitually employed in the manufactu...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Blackwell Publishing Ltd
2008
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3815885/ https://www.ncbi.nlm.nih.gov/pubmed/21261843 http://dx.doi.org/10.1111/j.1751-7915.2008.00027.x |
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author | Garmendia, Junkal De Las Heras, Aitor Galvão, Teca Calcagno De Lorenzo, Víctor |
author_facet | Garmendia, Junkal De Las Heras, Aitor Galvão, Teca Calcagno De Lorenzo, Víctor |
author_sort | Garmendia, Junkal |
collection | PubMed |
description | Although different biological approaches for detection of anti‐personnel mines and other unexploded ordnance (UXO) have been entertained, none of them has been rigorously documented thus far in the scientific literature. The industrial 2,4,6 trinitrotoluene (TNT) habitually employed in the manufacturing of mines is at all times tainted with a small but significant proportion of the more volatile 2,4 dinitrotoluene (2,4 DNT) and other nitroaromatic compounds. By using mutation‐prone PCR and DNA sequence shuffling we have evolved in vitro and selected in vivo variants of the effector recognition domain of the toluene‐responsive XylR regulator of the soil bacterium Pseudomonas putida that responds to mono‐, bi‐ and trinitro substituted toluenes. Re‐introduction of such variants in P. putida settled the transcriptional activity of the cognate promoters (Po and Pu) as a function of the presence of nitrotoluenes in the medium. When strains bearing transcriptional fusions to reporters with an optical output (luxAB, GFP) were spread on soil spotted with nitrotoluenes, the signal triggered by promoter activation allowed localization of the target compounds on the soil surface. Our data provide a proof of concept that non‐natural transcription factors evolved to respond to nitroaromatics can be engineered in soil bacteria and inoculated on a target site to pinpoint the presence of explosives. This approach thus opens new ways to tackle this gigantic humanitarian problem. |
format | Online Article Text |
id | pubmed-3815885 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2008 |
publisher | Blackwell Publishing Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-38158852014-02-12 Tracing explosives in soil with transcriptional regulators of Pseudomonas putida evolved for responding to nitrotoluenes Garmendia, Junkal De Las Heras, Aitor Galvão, Teca Calcagno De Lorenzo, Víctor Microb Biotechnol Research Articles Although different biological approaches for detection of anti‐personnel mines and other unexploded ordnance (UXO) have been entertained, none of them has been rigorously documented thus far in the scientific literature. The industrial 2,4,6 trinitrotoluene (TNT) habitually employed in the manufacturing of mines is at all times tainted with a small but significant proportion of the more volatile 2,4 dinitrotoluene (2,4 DNT) and other nitroaromatic compounds. By using mutation‐prone PCR and DNA sequence shuffling we have evolved in vitro and selected in vivo variants of the effector recognition domain of the toluene‐responsive XylR regulator of the soil bacterium Pseudomonas putida that responds to mono‐, bi‐ and trinitro substituted toluenes. Re‐introduction of such variants in P. putida settled the transcriptional activity of the cognate promoters (Po and Pu) as a function of the presence of nitrotoluenes in the medium. When strains bearing transcriptional fusions to reporters with an optical output (luxAB, GFP) were spread on soil spotted with nitrotoluenes, the signal triggered by promoter activation allowed localization of the target compounds on the soil surface. Our data provide a proof of concept that non‐natural transcription factors evolved to respond to nitroaromatics can be engineered in soil bacteria and inoculated on a target site to pinpoint the presence of explosives. This approach thus opens new ways to tackle this gigantic humanitarian problem. Blackwell Publishing Ltd 2008-05 2008-02-19 /pmc/articles/PMC3815885/ /pubmed/21261843 http://dx.doi.org/10.1111/j.1751-7915.2008.00027.x Text en Copyright © 2008 The Authors. Journal compilation © 2008 Society for Applied Microbiology and Blackwell Publishing Ltd. |
spellingShingle | Research Articles Garmendia, Junkal De Las Heras, Aitor Galvão, Teca Calcagno De Lorenzo, Víctor Tracing explosives in soil with transcriptional regulators of Pseudomonas putida evolved for responding to nitrotoluenes |
title | Tracing explosives in soil with transcriptional regulators of Pseudomonas putida evolved for responding to nitrotoluenes |
title_full | Tracing explosives in soil with transcriptional regulators of Pseudomonas putida evolved for responding to nitrotoluenes |
title_fullStr | Tracing explosives in soil with transcriptional regulators of Pseudomonas putida evolved for responding to nitrotoluenes |
title_full_unstemmed | Tracing explosives in soil with transcriptional regulators of Pseudomonas putida evolved for responding to nitrotoluenes |
title_short | Tracing explosives in soil with transcriptional regulators of Pseudomonas putida evolved for responding to nitrotoluenes |
title_sort | tracing explosives in soil with transcriptional regulators of pseudomonas putida evolved for responding to nitrotoluenes |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3815885/ https://www.ncbi.nlm.nih.gov/pubmed/21261843 http://dx.doi.org/10.1111/j.1751-7915.2008.00027.x |
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