Heterologous Expression of Pseudomonas putida Methyl-Accepting Chemotaxis Proteins Yields Escherichia coli Cells Chemotactic to Aromatic Compounds.

Détails

ID Serval
serval:BIB_3E2FA14125EB
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Heterologous Expression of Pseudomonas putida Methyl-Accepting Chemotaxis Proteins Yields Escherichia coli Cells Chemotactic to Aromatic Compounds.
Périodique
Applied and Environmental Microbiology
Auteur(s)
Roggo C., Clerc E.E., Hadadi N., Carraro N., Stocker R., van der Meer J.R.
ISSN
1098-5336 (Electronic)
ISSN-L
0099-2240
Statut éditorial
Publié
Date de publication
2018
Peer-reviewed
Oui
Volume
84
Numéro
18
Pages
UNSP e01362-18
Langue
anglais
Résumé
<i>Escherichia coli</i> , commonly used in chemotaxis studies, is attracted mostly by amino acids, sugars, and peptides. We envisioned modifying the chemotaxis specificity of <i>E. coli</i> by expressing heterologous chemoreceptors from <i>Pseudomonas putida</i> enabling attraction either to toluene or benzoate. The <i>mcpT</i> gene encoding the type 40-helical bundle (40H) methyl-accepting chemoreceptor for toluene from <i>Pseudomonas putida</i> MT53 and the <i>pcaY</i> gene for the type 40H receptor for benzoate and related molecules from <i>P. putida</i> F1 were expressed from the <i>trg</i> promoter on a plasmid in motile wild-type <i>E. coli</i> MG1655. <i>E. coli</i> cells expressing McpT accumulated in chemoattraction assays to sources with 60 to 200 μM toluene, although less strongly than the response to 100 μM serine, but statistically significantly stronger than that to sources without any added attractant. An McpT-mCherry fusion protein was detectably expressed in <i>E. coli</i> and yielded weak but distinguishable membranes and polar foci in 1% of cells. <i>E. coli</i> cells expressing PcaY showed weak attraction to 0.1 to 1 mM benzoate, but 50 to 70% of cells localized the PcaY-mCherry fusion to their membrane. We conclude that implementing heterologous receptors in the <i>E. coli</i> chemotaxis network is possible and, upon improvement of the compatibility of the type 40H chemoreceptors, may bear interest for biosensing. <b>IMPORTANCE</b> Bacterial chemotaxis might be harnessed for the development of rapid biosensors, in which chemical availability is deduced from cell accumulation to chemoattractants over time. Chemotaxis of <i>Escherichia coli</i> has been well studied, but the bacterium is not attracted to chemicals of environmental concern, such as aromatic solvents. We show here that heterologous chemoreceptors for aromatic compounds from <i>Pseudomonas putida</i> at least partly functionally complement the <i>E. coli</i> chemotaxis network, yielding cells attracted to toluene or benzoate. Complementation was still inferior to native chemoattractants, like serine, but our study demonstrates the potential for obtaining selective sensing for aromatic compounds in <i>E. coli</i> .
Mots-clé
biosensing, chemotaxis
Pubmed
Web of science
Open Access
Oui
Création de la notice
31/07/2018 16:22
Dernière modification de la notice
20/08/2019 14:34
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