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Applied Microbiology and Biotechnology

, Volume 98, Issue 15, pp 6701–6713 | Cite as

Enhancing recombinant protein production with an Escherichia coli host strain lacking insertion sequences

  • Myung Keun Park
  • Sang Hee Lee
  • Kyung Seok Yang
  • Suk-Chae Jung
  • Jun Hyoung LeeEmail author
  • Sun Chang KimEmail author
Applied genetics and molecular biotechnology

Abstract

The genomic stability and integrity of host strains are critical for the production of recombinant proteins in biotechnology. Bacterial genomes contain numerous jumping genetic elements, the insertion sequences (ISs) that cause a variety of genetic rearrangements, resulting in adverse effects such as genome and recombinant plasmid instability. To minimize the harmful effects of ISs on the expression of recombinant proteins in Escherichia coli, we developed an IS-free, minimized E. coli strain (MS56) in which about 23 % of the genome, including all ISs and many unnecessary genes, was removed. Here, we compared the expression profiles of recombinant proteins such as tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and bone morphogenetic protein-2 (BMP2) in MG1655 and MS56. Hopping of ISs (IS1, IS3, or IS5) into the TRAIL and BMP2 genes occurred at the rate of ~10−8/gene/h in MG1655 whereas such events were not observed in MS56. Even though IS hopping occurred very rarely (10−8/gene/h), cells containing the IS-inserted TRAIL and BMP2 plasmids became dominant (~52 % of the total population) 28 h after fermentation began due to their growth advantage over cells containing intact plasmids, significantly reducing recombinant protein production in batch fermentation. Our findings clearly indicate that IS hopping is detrimental to the industrial production of recombinant proteins, emphasizing the importance of the development of IS-free host strains.

Keywords

Insertion sequence IS hopping E. coli Genomic stability 

Notes

Acknowledgments

This work was supported by grants from the Intelligent Synthetic Biology Center of Global Frontier Project funded by the Ministry of Science, ICT and Future Planning (2011–0031955) and the Next-Generation BioGreen 21 Program (SSAC; PJ008110). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Supplementary material

253_2014_5739_MOESM1_ESM.pdf (57 kb)
ESM 1 (PDF 56 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Myung Keun Park
    • 1
  • Sang Hee Lee
    • 1
  • Kyung Seok Yang
    • 1
  • Suk-Chae Jung
    • 1
  • Jun Hyoung Lee
    • 2
    Email author
  • Sun Chang Kim
    • 1
    Email author
  1. 1.Department of Biological SciencesKAISTDaejeonSouth Korea
  2. 2.KAIST Institute for BiocenturyKAISTDaejeonSouth Korea

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