Molecular and Cellular Biochemistry

, Volume 307, Issue 1–2, pp 249–264 | Cite as

Production of active eukaryotic proteins through bacterial expression systems: a review of the existing biotechnology strategies

  • Sudhir Sahdev
  • Sunil K. Khattar
  • Kulvinder Singh Saini
Article

Abstract

Among the various expression systems employed for the over-production of proteins, bacteria still remains the favorite choice of a Protein Biochemist. However, even today, due to the lack of post-translational modification machinery in bacteria, recombinant eukaryotic protein production poses an immense challenge, which invariably leads to the production of biologically in-active protein in this host. A number of techniques are cited in the literature, which describe the conversion of inactive protein, expressed as an insoluble fraction, into a soluble and active form. Overall, we have divided these methods into three major groups: Group-I, where the factors influencing the formation of insoluble fraction are modified through a stringent control of the cellular milieu, thereby leading to the expression of recombinant protein as soluble moiety; Group-II, where protein is refolded from the inclusion bodies and thereby target protein modification is avoided; Group-III, where the target protein is engineered to achieve soluble expression through fusion protein technology. Even within the same family of proteins (e.g., tyrosine kinases), optimization of standard operating protocol (SOP) may still be required for each protein’s over-production at a pilot-scale in Escherichia coli. However, once standardized, this procedure can be made amenable to the industrial production for that particular protein with minimum alterations.

Keywords

Post-translational modifications Disulfide bond Phosphorylation Glycosylation Inclusion body refolding Size exclusion chromatography Hydrophobic interactions 

Notes

Acknowledgments

This work was funded by Ranbaxy Research Laboratories, India. We thankfully acknowledge our colleagues within the New Drug Discovery Research Group, Department of Biotechnology Team at Ranbaxy, particularly Dr. R.S. Bora, Dr. R. Kant, Mr. Roop Trivedi for their assistance during review compilation and Mr. Prabhakar Tiwari, Ms. Shalini Saini for generating the unpublished data.

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Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Sudhir Sahdev
    • 1
  • Sunil K. Khattar
    • 1
  • Kulvinder Singh Saini
    • 1
  1. 1.Department of Biotechnology & Bioinformatics, New Drug Discovery ResearchRanbaxy Research Laboratories-R&D-3GurgaonIndia

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