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

• Main Authors: Luciana Gomes, Gabriel Monteiro, Filipe Mergulhão
• Format: Article
• Language: English
• Published: MDPI AG 2020-01-01
• Series: International Journal of Molecular Sciences
• Subjects: biofilm; <i>escherichia coli</i>; heterologous protein expression; plasmid stability; plasmid copy number; iptg
• Online Access: https://www.mdpi.com/1422-0067/21/2/576

This work assesses the effect of chemical induction with isopropyl &#946;-D-1-thiogalactopyranoside (IPTG) on the expression of enhanced green fluorescent protein (eGFP) by planktonic and biofilm cells of <i>Escherichia coli</i> 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.