Cargando…

Development and application of a transcriptional sensor for detection of heterologous acrylic acid production in E. coli

BACKGROUND: Acrylic acid (AA) is a widely used commodity chemical derived from non-renewable fossil fuel sources. Alternative microbial-based production methodologies are being developed with the aim of providing “green” acrylic acid. These initiatives will benefit from component sensing tools that...

Descripción completa

Detalles Bibliográficos
Autores principales: Raghavan, Sarada S., Chee, Sharon, Li, Juntao, Poschmann, Jeremie, Nagarajan, Niranjan, Jia Wei, Siau, Verma, Chandra S., Ghadessy, Farid J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6699081/
https://www.ncbi.nlm.nih.gov/pubmed/31426802
http://dx.doi.org/10.1186/s12934-019-1185-y
_version_ 1783444658575114240
author Raghavan, Sarada S.
Chee, Sharon
Li, Juntao
Poschmann, Jeremie
Nagarajan, Niranjan
Jia Wei, Siau
Verma, Chandra S.
Ghadessy, Farid J.
author_facet Raghavan, Sarada S.
Chee, Sharon
Li, Juntao
Poschmann, Jeremie
Nagarajan, Niranjan
Jia Wei, Siau
Verma, Chandra S.
Ghadessy, Farid J.
author_sort Raghavan, Sarada S.
collection PubMed
description BACKGROUND: Acrylic acid (AA) is a widely used commodity chemical derived from non-renewable fossil fuel sources. Alternative microbial-based production methodologies are being developed with the aim of providing “green” acrylic acid. These initiatives will benefit from component sensing tools that facilitate rapid and easy detection of in vivo AA production. RESULTS: We developed a novel transcriptional sensor facilitating in vivo detection of acrylic acid (AA). RNAseq analysis of Escherichia coli exposed to sub-lethal doses of acrylic acid identified a selectively responsive promoter (P(yhcN)) that was cloned upstream of the eGFP gene. In the presence of AA, eGFP expression in E. coli cells harbouring the sensing construct was readily observable by fluorescence read-out. Low concentrations of AA (500 μM) could be detected whilst the closely related lactic and 3-hydroxy propionic acids failed to activate the sensor. We further used the developed AA-biosensor for in vivo FACS-based screening and identification of amidase mutants with improved catalytic properties for deamination of acrylamide to acrylic acid. CONCLUSIONS: The transcriptional AA sensor developed in this study will benefit strain, enzyme and pathway engineering initiatives targeting the efficient formation of bio-acrylic acid. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12934-019-1185-y) contains supplementary material, which is available to authorized users.
format Online
Article
Text
id pubmed-6699081
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-66990812019-08-26 Development and application of a transcriptional sensor for detection of heterologous acrylic acid production in E. coli Raghavan, Sarada S. Chee, Sharon Li, Juntao Poschmann, Jeremie Nagarajan, Niranjan Jia Wei, Siau Verma, Chandra S. Ghadessy, Farid J. Microb Cell Fact Research BACKGROUND: Acrylic acid (AA) is a widely used commodity chemical derived from non-renewable fossil fuel sources. Alternative microbial-based production methodologies are being developed with the aim of providing “green” acrylic acid. These initiatives will benefit from component sensing tools that facilitate rapid and easy detection of in vivo AA production. RESULTS: We developed a novel transcriptional sensor facilitating in vivo detection of acrylic acid (AA). RNAseq analysis of Escherichia coli exposed to sub-lethal doses of acrylic acid identified a selectively responsive promoter (P(yhcN)) that was cloned upstream of the eGFP gene. In the presence of AA, eGFP expression in E. coli cells harbouring the sensing construct was readily observable by fluorescence read-out. Low concentrations of AA (500 μM) could be detected whilst the closely related lactic and 3-hydroxy propionic acids failed to activate the sensor. We further used the developed AA-biosensor for in vivo FACS-based screening and identification of amidase mutants with improved catalytic properties for deamination of acrylamide to acrylic acid. CONCLUSIONS: The transcriptional AA sensor developed in this study will benefit strain, enzyme and pathway engineering initiatives targeting the efficient formation of bio-acrylic acid. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12934-019-1185-y) contains supplementary material, which is available to authorized users. BioMed Central 2019-08-19 /pmc/articles/PMC6699081/ /pubmed/31426802 http://dx.doi.org/10.1186/s12934-019-1185-y Text en © The Author(s) 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Raghavan, Sarada S.
Chee, Sharon
Li, Juntao
Poschmann, Jeremie
Nagarajan, Niranjan
Jia Wei, Siau
Verma, Chandra S.
Ghadessy, Farid J.
Development and application of a transcriptional sensor for detection of heterologous acrylic acid production in E. coli
title Development and application of a transcriptional sensor for detection of heterologous acrylic acid production in E. coli
title_full Development and application of a transcriptional sensor for detection of heterologous acrylic acid production in E. coli
title_fullStr Development and application of a transcriptional sensor for detection of heterologous acrylic acid production in E. coli
title_full_unstemmed Development and application of a transcriptional sensor for detection of heterologous acrylic acid production in E. coli
title_short Development and application of a transcriptional sensor for detection of heterologous acrylic acid production in E. coli
title_sort development and application of a transcriptional sensor for detection of heterologous acrylic acid production in e. coli
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6699081/
https://www.ncbi.nlm.nih.gov/pubmed/31426802
http://dx.doi.org/10.1186/s12934-019-1185-y
work_keys_str_mv AT raghavansaradas developmentandapplicationofatranscriptionalsensorfordetectionofheterologousacrylicacidproductioninecoli
AT cheesharon developmentandapplicationofatranscriptionalsensorfordetectionofheterologousacrylicacidproductioninecoli
AT lijuntao developmentandapplicationofatranscriptionalsensorfordetectionofheterologousacrylicacidproductioninecoli
AT poschmannjeremie developmentandapplicationofatranscriptionalsensorfordetectionofheterologousacrylicacidproductioninecoli
AT nagarajanniranjan developmentandapplicationofatranscriptionalsensorfordetectionofheterologousacrylicacidproductioninecoli
AT jiaweisiau developmentandapplicationofatranscriptionalsensorfordetectionofheterologousacrylicacidproductioninecoli
AT vermachandras developmentandapplicationofatranscriptionalsensorfordetectionofheterologousacrylicacidproductioninecoli
AT ghadessyfaridj developmentandapplicationofatranscriptionalsensorfordetectionofheterologousacrylicacidproductioninecoli