Proteomics and metabolic burden analysis to understand the impact of recombinant protein production in E. coli

Sci Rep. 2024 May 28;14(1):12271. doi: 10.1038/s41598-024-63148-y.

Abstract

The impact of recombinant protein production (RPP) on host cells and the metabolic burden associated with it undermine the efficiency of the production system. This study utilized proteomics to investigate the dynamics of parent and recombinant cells induced at different time points for RPP. The results revealed significant changes in both transcriptional and translational machinery that may have impacted the metabolic burden, growth rate of the culture and the RPP. The timing of protein synthesis induction also played a critical role in the fate of the recombinant protein within the host cell, affecting protein and product yield. The study identified significant differences in the expression of proteins involved in fatty acid and lipid biosynthesis pathways between two E. coli host strains (M15 and DH5⍺), with the E. coli M15 strain demonstrating superior expression characteristics for the recombinant protein. Overall, these findings contribute to the knowledge base for rational strain engineering for optimized recombinant protein production.

Keywords: Acyl-(acyl carrier protein) reductase (AAR); Gene expression; Metabolic burden; Post-induction growth phases; Proteomics; Recombinant protein production.

MeSH terms

  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism
  • Escherichia coli* / genetics
  • Escherichia coli* / metabolism
  • Fatty Acids / biosynthesis
  • Fatty Acids / metabolism
  • Protein Biosynthesis
  • Proteomics* / methods
  • Recombinant Proteins* / genetics
  • Recombinant Proteins* / metabolism

Substances

  • Recombinant Proteins
  • Escherichia coli Proteins
  • Fatty Acids