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Review

Review

Production of Biopharmaceuticals in E. coli: Current Scenario and Future Perspectives

Mohammed N. Baeshen 1, Ahmed M. Al-Hejin 1, Roop S. Bora 1, 2, Mohamed M. M. Ahmed 1, 3*, Hassan A. I. Ramadan 1, 4, Kulvinder S. Saini 1, 2, Nabih A. Baeshen 1 and Elrashdy M. Redwan 1, 5

1Department of Biological Sciences, Faculty of Science, King Abdulaziz University, Jeddah, 21589, Saudi Arabia, 2Department of Biotechnology, Eternal University, Baru Sahib-173 101, Himachal Pradesh, India, 3Nucleic Acids Research Department, Genetic Engineering and Biotechnology Research Institute (GEBRI), City for Scientific Research and Technology Applications, Alexandria 21934, Egypt, 4Cell Biology Department, Genetic Engineering and Biotechnology Division, National Research Centre, Dokki-Cairo 12311, Egypt, 5Protein Research Department, Genetic Engineering and Biotechnology Research Institute, City for Scientific Research and Applied Technology, New Borg AL-Arab, Alexandria 21934, Egypt

Received: December 30, 2014; Accepted: February 13, 2015

J. Microbiol. Biotechnol. 2015; 25(7): 953-962

Published July 28, 2015 https://doi.org/10.4014/jmb.1412.12079

Copyright © The Korean Society for Microbiology and Biotechnology.

Abstract

Escherichia coli is the most preferred microorganism to express heterologous proteins for therapeutic use, as around 30% of the approved therapeutic proteins are currently being produced using it as a host. Owing to its rapid growth, high yield of the product, costeffectiveness, and easy scale-up process, E. coli is an expression host of choice in the biotechnology industry for large-scale production of proteins, particularly non-glycosylated proteins, for therapeutic use. The availability of various E. coli expression vectors and strains, relatively easy protein folding mechanisms, and bioprocess technologies, makes it very attractive for industrial applications. However, the codon usage in E. coli and the absence of post-translational modifications, such as glycosylation, phosphorylation, and proteolytic processing, limit its use for the production of slightly complex recombinant biopharmaceuticals. Several new technological advancements in the E. coli expression system to meet the biotechnology industry requirements have been made, such as novel engineered strains, genetically modifying E. coli to possess capability to glycosylate heterologous proteins and express complex proteins, including full-length glycosylated antibodies. This review summarizes the recent advancements that may further expand the use of the E. coli expression system to produce more complex and also glycosylated proteins for therapeutic use in the future.

Keywords

E. coli, Optimized protein production, Biopharmaceuticals, Codon usage, Molecular chaperones

References

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    CrossRef
  2. Betiku E. 2006. Molecular chaperones involved in heterologous protein folding in Escherichia coli. Biotechnol. Mol. Biol. 1: 66-75.
  3. Brinkman U, Mattes RE, Buckel P. 1989. High-level expression of recombinant genes in Escherichia coli i s dependent on the availability of the dnaY gene product. Gene 85: 109-114.
    CrossRef
  4. Calderone TL, Stevens RD, Oas TG. 1996. High-level misincorporation of lysine for arginine at AGA codons in a fusion protein expressed in Escherichia coli. J. Mol. Biol. 262:407-412.
    Pubmed CrossRef
  5. Carrio MM, Villaverde A. 2003. Role of molecular chaperones in inclusion body formation. FEBS Lett. 537: 215-221.
    CrossRef
  6. Chen R. 2012. Bacterial expression systems for recombinant protein production: E. coli and beyond. Biotechnol. Adv. 30: 1102-1107.
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  7. Choi JH, Lee SY. 2004. Secretory and extracellular production of recombinant proteins using Escherichia coli. Appl. Microbiol. Biotechnol. 64: 625-635.
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  8. Cuccui J, Wren B. 2015. Hijacking bacterial glycosylation for the production of glycoconjugates, from vaccines to humanised glycoproteins. J. Pharm. Pharmacol. 67: 338-350.
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  9. Cui SS, Lin XZ, Shen JH. 2011. Effects of co-expression of molecular chaperones on heterologous soluble expression of the cold-active lipase Lip-948. Protein Expr. Purif. 77: 166-172.
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Article

Review

J. Microbiol. Biotechnol. 2015; 25(7): 953-962

Published online July 28, 2015 https://doi.org/10.4014/jmb.1412.12079

Copyright © The Korean Society for Microbiology and Biotechnology.

Production of Biopharmaceuticals in E. coli: Current Scenario and Future Perspectives

Mohammed N. Baeshen 1, Ahmed M. Al-Hejin 1, Roop S. Bora 1, 2, Mohamed M. M. Ahmed 1, 3*, Hassan A. I. Ramadan 1, 4, Kulvinder S. Saini 1, 2, Nabih A. Baeshen 1 and Elrashdy M. Redwan 1, 5

1Department of Biological Sciences, Faculty of Science, King Abdulaziz University, Jeddah, 21589, Saudi Arabia, 2Department of Biotechnology, Eternal University, Baru Sahib-173 101, Himachal Pradesh, India, 3Nucleic Acids Research Department, Genetic Engineering and Biotechnology Research Institute (GEBRI), City for Scientific Research and Technology Applications, Alexandria 21934, Egypt, 4Cell Biology Department, Genetic Engineering and Biotechnology Division, National Research Centre, Dokki-Cairo 12311, Egypt, 5Protein Research Department, Genetic Engineering and Biotechnology Research Institute, City for Scientific Research and Applied Technology, New Borg AL-Arab, Alexandria 21934, Egypt

Received: December 30, 2014; Accepted: February 13, 2015

Abstract

Escherichia coli is the most preferred microorganism to express heterologous proteins for
therapeutic use, as around 30% of the approved therapeutic proteins are currently being
produced using it as a host. Owing to its rapid growth, high yield of the product, costeffectiveness,
and easy scale-up process, E. coli is an expression host of choice in the
biotechnology industry for large-scale production of proteins, particularly non-glycosylated
proteins, for therapeutic use. The availability of various E. coli expression vectors and strains,
relatively easy protein folding mechanisms, and bioprocess technologies, makes it very
attractive for industrial applications. However, the codon usage in E. coli and the absence of
post-translational modifications, such as glycosylation, phosphorylation, and proteolytic
processing, limit its use for the production of slightly complex recombinant biopharmaceuticals.
Several new technological advancements in the E. coli expression system to meet the
biotechnology industry requirements have been made, such as novel engineered strains,
genetically modifying E. coli to possess capability to glycosylate heterologous proteins and
express complex proteins, including full-length glycosylated antibodies. This review summarizes
the recent advancements that may further expand the use of the E. coli expression system to
produce more complex and also glycosylated proteins for therapeutic use in the future.

Keywords: E. coli, Optimized protein production, Biopharmaceuticals, Codon usage, Molecular chaperones

References

  1. Arya R, Sabir JSM, Bora RS, Saini KS. 2015. Optimization of culture parameters and novel strategies to improve protein solubility. In Insoluble Proteins: Methods and Protocols. Methods in Molecular Biology series, edited by Dr. Elena Garcia Fruitos, Vol. 1258, Chapter 3. Pages 45-63; Springer Science and Business Media, New York, USA.
    CrossRef
  2. Betiku E. 2006. Molecular chaperones involved in heterologous protein folding in Escherichia coli. Biotechnol. Mol. Biol. 1: 66-75.
  3. Brinkman U, Mattes RE, Buckel P. 1989. High-level expression of recombinant genes in Escherichia coli i s dependent on the availability of the dnaY gene product. Gene 85: 109-114.
    CrossRef
  4. Calderone TL, Stevens RD, Oas TG. 1996. High-level misincorporation of lysine for arginine at AGA codons in a fusion protein expressed in Escherichia coli. J. Mol. Biol. 262:407-412.
    Pubmed CrossRef
  5. Carrio MM, Villaverde A. 2003. Role of molecular chaperones in inclusion body formation. FEBS Lett. 537: 215-221.
    CrossRef
  6. Chen R. 2012. Bacterial expression systems for recombinant protein production: E. coli and beyond. Biotechnol. Adv. 30: 1102-1107.
    Pubmed CrossRef
  7. Choi JH, Lee SY. 2004. Secretory and extracellular production of recombinant proteins using Escherichia coli. Appl. Microbiol. Biotechnol. 64: 625-635.
    Pubmed CrossRef
  8. Cuccui J, Wren B. 2015. Hijacking bacterial glycosylation for the production of glycoconjugates, from vaccines to humanised glycoproteins. J. Pharm. Pharmacol. 67: 338-350.
    Pubmed KoreaMed CrossRef
  9. Cui SS, Lin XZ, Shen JH. 2011. Effects of co-expression of molecular chaperones on heterologous soluble expression of the cold-active lipase Lip-948. Protein Expr. Purif. 77: 166-172.
    Pubmed CrossRef
  10. de Marco A. 2007. Protocol for preparing proteins with improved solubility by co-expressing with molecular chaperones in Escherichia coli. Nat. Protoc. 2: 2632-2639.
    Pubmed CrossRef
  11. De Smit MH, van Duin, J. 1990. Secondary structure of the ribosome binding site determines translational efficiency: a quantitative analysis. Proc. Natl. Acad. Sci. USA 87: 7668-7672.
    Pubmed KoreaMed CrossRef
  12. Dieci G, Bottarelli L, Ballabeni A, Ottonello S. 2000. tRNA assisted overproduction of eukaryotic ribosomal proteins. Protein Expr. Purif. 18: 346-354.
    Pubmed CrossRef
  13. El-Baky NA, Redwan EM. 2015. Therapeutic alpha-interferons protein: structure, production, and biosimilar. Prep. Biochem. Biotechnol. 45: 109-127.
    Pubmed CrossRef
  14. Etchegaray JP, Inouye M. 1999. Translational enhancement by an element example of molecular misreading in Alzheimer’s disease. Trends Neurosci. 21: 331-335.
  15. Fahnert B, Lilie H, Neubauer P. 2004. Inclusion bodies:formation and utilization. Adv. Biochem. Eng. Biotechnol. 89:93-142.
    Pubmed CrossRef
  16. Feng Y, Xu Q, Yang T, Sun E, Li J, Shi D, Wu D. 2014. A novel self-cleavage system for production of soluble recombinant protein in Escherichia coli. Protein Expr. Purif. 99: 64-69.
    Pubmed CrossRef
  17. Ferrer-Miralles N, Villaverde A. 2013. Bacterial cell factories for recombinant protein production; expanding the catalogue. Microb. Cell Fact. 12: 113.
    Pubmed KoreaMed CrossRef
  18. Fisher AC, Haitjema CH, Guarino C, Celik E, Endicott CE,Reading CA, et al. 2011. Production of secretory and extracellular N-linked glycoproteins in Escherichia coli. Appl. Environ. Microbiol. 77: 871-881.
    Pubmed KoreaMed CrossRef
  19. Folwarczna J, Moravec T, Plchova H, Hoffmeisterova H, Cerovska N. 2012. Efficient expression of human papillomavirus 16 E7 oncoprotein fused to C-terminus of tobacco mosaic virus (TMV) coat protein using molecular chaperones in Escherichia coli. Protein Expr. Purif. 85: 152-157.
    Pubmed CrossRef
  20. Guzzo J. 2012. Biotechnical applications of small heat shock proteins from bacteria. Int. J. Biochem. Cell Biol. 44: 1698-1705.
    Pubmed CrossRef
  21. Ihss en J , Kowarik M, D ilettos o S, T anner C, W acker M , Thöny-Meyer L. 2010. Production of glycoprotein vaccines in Escherichia coli. Microb. Cell Fact. 11: 9: 61.
  22. Iost I, Dreyfus M. 1994. mRNAs can be stabilized by DEADbox proteins. Nature 372: 193-196.
    Pubmed CrossRef
  23. Iost I, Bizebard T, Dreyfus M. 2013. Functions of DEAD-box proteins in bacteria: current knowledge and pending questions. Biochim. Biophys. Acta 1829: 866-877.
    Pubmed CrossRef
  24. Jensen EB, Carlsen S. 1990. Production of recombinant human growth hormone in Escherichia coli: expression of different precursors and physiological effects of glucose, acetate and salts. Biotechnol. Bioeng. 36: 1-11.
    Pubmed CrossRef
  25. Jenkins N. 2007. Modifications of therapeutic proteins:challenges and prospects. Cytotechnology 53: 121-125.
    Pubmed KoreaMed CrossRef
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