Cargando…

The Impact of IPTG Induction on Plasmid Stability and Heterologous Protein Expression by Escherichia coli Biofilms

This work assesses the effect of chemical induction with isopropyl β-D-1-thiogalactopyranoside (IPTG) on the expression of enhanced green fluorescent protein (eGFP) by planktonic and biofilm cells of Escherichia coli JM109(DE3) transformed with a plasmid containing a T7 promoter. It was shown that i...

Descripción completa

Detalles Bibliográficos
Autores principales: Gomes, Luciana, Monteiro, Gabriel, Mergulhão, Filipe
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7013871/
https://www.ncbi.nlm.nih.gov/pubmed/31963160
http://dx.doi.org/10.3390/ijms21020576
_version_ 1783496498263097344
author Gomes, Luciana
Monteiro, Gabriel
Mergulhão, Filipe
author_facet Gomes, Luciana
Monteiro, Gabriel
Mergulhão, Filipe
author_sort Gomes, Luciana
collection PubMed
description This work assesses the effect of chemical induction with isopropyl β-D-1-thiogalactopyranoside (IPTG) on the expression of enhanced green fluorescent protein (eGFP) by planktonic and biofilm cells of Escherichia coli JM109(DE3) transformed with a plasmid containing a T7 promoter. It was shown that induction negatively affected the growth and viability of planktonic cultures, and eGFP production did not increase. Heterologous protein production was not limited by gene dosage or by transcriptional activity. Results suggest that plasmid maintenance at high copy number imposes a metabolic burden that precludes high level expression of the heterologous protein. In biofilm cells, the inducer avoided the overall decrease in the amount of expressed eGFP, although this was not correlated with the gene dosage. Higher specific production levels were always attained with biofilm cells and it seems that while induction of biofilm cells shifts their metabolism towards the maintenance of heterologous protein concentration, in planktonic cells the cellular resources are directed towards plasmid replication and growth.
format Online
Article
Text
id pubmed-7013871
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-70138712020-03-09 The Impact of IPTG Induction on Plasmid Stability and Heterologous Protein Expression by Escherichia coli Biofilms Gomes, Luciana Monteiro, Gabriel Mergulhão, Filipe Int J Mol Sci Article This work assesses the effect of chemical induction with isopropyl β-D-1-thiogalactopyranoside (IPTG) on the expression of enhanced green fluorescent protein (eGFP) by planktonic and biofilm cells of Escherichia coli JM109(DE3) transformed with a plasmid containing a T7 promoter. It was shown that induction negatively affected the growth and viability of planktonic cultures, and eGFP production did not increase. Heterologous protein production was not limited by gene dosage or by transcriptional activity. Results suggest that plasmid maintenance at high copy number imposes a metabolic burden that precludes high level expression of the heterologous protein. In biofilm cells, the inducer avoided the overall decrease in the amount of expressed eGFP, although this was not correlated with the gene dosage. Higher specific production levels were always attained with biofilm cells and it seems that while induction of biofilm cells shifts their metabolism towards the maintenance of heterologous protein concentration, in planktonic cells the cellular resources are directed towards plasmid replication and growth. MDPI 2020-01-16 /pmc/articles/PMC7013871/ /pubmed/31963160 http://dx.doi.org/10.3390/ijms21020576 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Gomes, Luciana
Monteiro, Gabriel
Mergulhão, Filipe
The Impact of IPTG Induction on Plasmid Stability and Heterologous Protein Expression by Escherichia coli Biofilms
title The Impact of IPTG Induction on Plasmid Stability and Heterologous Protein Expression by Escherichia coli Biofilms
title_full The Impact of IPTG Induction on Plasmid Stability and Heterologous Protein Expression by Escherichia coli Biofilms
title_fullStr The Impact of IPTG Induction on Plasmid Stability and Heterologous Protein Expression by Escherichia coli Biofilms
title_full_unstemmed The Impact of IPTG Induction on Plasmid Stability and Heterologous Protein Expression by Escherichia coli Biofilms
title_short The Impact of IPTG Induction on Plasmid Stability and Heterologous Protein Expression by Escherichia coli Biofilms
title_sort impact of iptg induction on plasmid stability and heterologous protein expression by escherichia coli biofilms
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7013871/
https://www.ncbi.nlm.nih.gov/pubmed/31963160
http://dx.doi.org/10.3390/ijms21020576
work_keys_str_mv AT gomesluciana theimpactofiptginductiononplasmidstabilityandheterologousproteinexpressionbyescherichiacolibiofilms
AT monteirogabriel theimpactofiptginductiononplasmidstabilityandheterologousproteinexpressionbyescherichiacolibiofilms
AT mergulhaofilipe theimpactofiptginductiononplasmidstabilityandheterologousproteinexpressionbyescherichiacolibiofilms
AT gomesluciana impactofiptginductiononplasmidstabilityandheterologousproteinexpressionbyescherichiacolibiofilms
AT monteirogabriel impactofiptginductiononplasmidstabilityandheterologousproteinexpressionbyescherichiacolibiofilms
AT mergulhaofilipe impactofiptginductiononplasmidstabilityandheterologousproteinexpressionbyescherichiacolibiofilms