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Amino acid misincorporation during high-level expression of mouse epidermal growth factor in Escherichia coli

C A Scorer1, M J Carrier, R F Rosenberger

  • 1Genetics Division, National Institute for Medical Research, London, UK.

Nucleic Acids Research
|July 11, 1991
PubMed

Insights

High-level protein production in Escherichia coli causes frequent translational errors, with an error rate 10x higher than normal. This impacts the clinical use of E. coli-derived therapeutic proteins.

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Protein Synthesis

Background:

  • Escherichia coli is a common host for producing foreign proteins.
  • High-level expression of heterologous proteins can potentially disrupt cellular processes.

Purpose of the Study:

  • To investigate translational errors during high-level foreign protein production in E. coli.
  • To quantify amino acid misincorporation in mouse epidermal growth factor (mEGF).

Main Methods:

  • Produced mEGF as a TrpE fusion protein in E. coli.
  • Analyzed phenylalanine content in purified protein to measure missense errors, as mEGF DNA lacks phenylalanine codons.

Main Results:

  • Identified an amino acid misincorporation error frequency of approximately 1 in 40 for phenylalanine codons.
  • This error rate is at least ten times higher than typical E. coli protein synthesis.
  • Hypothesized that limited charged tRNAs and GTP contribute to errors.

Conclusions:

  • High-level heterologous protein expression in E. coli leads to significant translational fidelity loss.
  • The observed high error rate has critical implications for the safety and efficacy of therapeutic proteins produced in E. coli.

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