Genome-wide identification of tolerance mechanisms towards p-coumaric acid in Pseudomonas putida

Patricia Calero, Sheila I. Jensen, Klara Bojanovič, Rebecca Lennen, Anna Koza, Alex Toftgaard Nielsen*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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Abstract

The soil bacterium Pseudomonas putida KT2440 has gained increasing biotechnological interest due to its ability to tolerate different types of stress. Here, the tolerance of P. putida KT2440 towards eleven toxic chemical compounds was investigated. P. putida was found to be significantly more tolerant towards three of the eleven compounds when compared to Escherichia coli. Increased tolerance was for example found towards p-coumaric acid, an interesting precursor for polymerization with a significant industrial relevance. The tolerance mechanism was therefore investigated using the genome-wide approach, Tn-seq. Libraries containing a large number of miniTn5-Km transposon insertion mutants were grown in the presence and absence of p-coumaric acid, and the enrichment or depletion of mutants was quantified by high-throughput sequencing. Several genes, including the ABC transporter Ttg2ABC and the cytochrome c maturation system (ccm), were identified to play an important role in the tolerance towards p-coumaric acid of this bacterium. Most of the identified genes were involved in membrane stability, suggesting that tolerance towards p-coumaric acid is related to transport and membrane integrity. This article is protected by copyright. All rights reserved.
Original languageEnglish
JournalBiotechnology and Bioengineering
Volume115
Issue number3
Pages (from-to)762-774
ISSN0006-3592
DOIs
Publication statusPublished - 2018

Bibliographical note

This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

Keywords

  • Pseudomonas putida
  • Tn-seq
  • p-Coumaric acid
  • Tolerance

Cite this

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title = "Genome-wide identification of tolerance mechanisms towards p-coumaric acid in Pseudomonas putida",
abstract = "The soil bacterium Pseudomonas putida KT2440 has gained increasing biotechnological interest due to its ability to tolerate different types of stress. Here, the tolerance of P. putida KT2440 towards eleven toxic chemical compounds was investigated. P. putida was found to be significantly more tolerant towards three of the eleven compounds when compared to Escherichia coli. Increased tolerance was for example found towards p-coumaric acid, an interesting precursor for polymerization with a significant industrial relevance. The tolerance mechanism was therefore investigated using the genome-wide approach, Tn-seq. Libraries containing a large number of miniTn5-Km transposon insertion mutants were grown in the presence and absence of p-coumaric acid, and the enrichment or depletion of mutants was quantified by high-throughput sequencing. Several genes, including the ABC transporter Ttg2ABC and the cytochrome c maturation system (ccm), were identified to play an important role in the tolerance towards p-coumaric acid of this bacterium. Most of the identified genes were involved in membrane stability, suggesting that tolerance towards p-coumaric acid is related to transport and membrane integrity. This article is protected by copyright. All rights reserved.",
keywords = "Pseudomonas putida, Tn-seq, p-Coumaric acid, Tolerance",
author = "Patricia Calero and Jensen, {Sheila I.} and Klara Bojanovič and Rebecca Lennen and Anna Koza and Nielsen, {Alex Toftgaard}",
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Genome-wide identification of tolerance mechanisms towards p-coumaric acid in Pseudomonas putida. / Calero, Patricia; Jensen, Sheila I.; Bojanovič, Klara; Lennen, Rebecca; Koza, Anna; Nielsen, Alex Toftgaard.

In: Biotechnology and Bioengineering, Vol. 115, No. 3, 2018, p. 762-774.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - Genome-wide identification of tolerance mechanisms towards p-coumaric acid in Pseudomonas putida

AU - Calero, Patricia

AU - Jensen, Sheila I.

AU - Bojanovič, Klara

AU - Lennen, Rebecca

AU - Koza, Anna

AU - Nielsen, Alex Toftgaard

N1 - This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

PY - 2018

Y1 - 2018

N2 - The soil bacterium Pseudomonas putida KT2440 has gained increasing biotechnological interest due to its ability to tolerate different types of stress. Here, the tolerance of P. putida KT2440 towards eleven toxic chemical compounds was investigated. P. putida was found to be significantly more tolerant towards three of the eleven compounds when compared to Escherichia coli. Increased tolerance was for example found towards p-coumaric acid, an interesting precursor for polymerization with a significant industrial relevance. The tolerance mechanism was therefore investigated using the genome-wide approach, Tn-seq. Libraries containing a large number of miniTn5-Km transposon insertion mutants were grown in the presence and absence of p-coumaric acid, and the enrichment or depletion of mutants was quantified by high-throughput sequencing. Several genes, including the ABC transporter Ttg2ABC and the cytochrome c maturation system (ccm), were identified to play an important role in the tolerance towards p-coumaric acid of this bacterium. Most of the identified genes were involved in membrane stability, suggesting that tolerance towards p-coumaric acid is related to transport and membrane integrity. This article is protected by copyright. All rights reserved.

AB - The soil bacterium Pseudomonas putida KT2440 has gained increasing biotechnological interest due to its ability to tolerate different types of stress. Here, the tolerance of P. putida KT2440 towards eleven toxic chemical compounds was investigated. P. putida was found to be significantly more tolerant towards three of the eleven compounds when compared to Escherichia coli. Increased tolerance was for example found towards p-coumaric acid, an interesting precursor for polymerization with a significant industrial relevance. The tolerance mechanism was therefore investigated using the genome-wide approach, Tn-seq. Libraries containing a large number of miniTn5-Km transposon insertion mutants were grown in the presence and absence of p-coumaric acid, and the enrichment or depletion of mutants was quantified by high-throughput sequencing. Several genes, including the ABC transporter Ttg2ABC and the cytochrome c maturation system (ccm), were identified to play an important role in the tolerance towards p-coumaric acid of this bacterium. Most of the identified genes were involved in membrane stability, suggesting that tolerance towards p-coumaric acid is related to transport and membrane integrity. This article is protected by copyright. All rights reserved.

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