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

Int J Mol Sci. 2020 Jan 16;21(2):576. doi: 10.3390/ijms21020576.

Abstract

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.

Keywords: Escherichia coli; IPTG; biofilm; heterologous protein expression; plasmid copy number; plasmid stability.

MeSH terms

  • Biofilms / growth & development*
  • Escherichia coli / growth & development*
  • Escherichia coli / metabolism*
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Isopropyl Thiogalactoside / metabolism
  • Plasmids / genetics
  • Plasmids / metabolism
  • Promoter Regions, Genetic / genetics

Substances

  • Escherichia coli Proteins
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Isopropyl Thiogalactoside