Skip to main content

Choose a Suitable Expression Host: A Survey of Available Protein Production Platforms

  • Chapter
  • First Online:
Advanced Technologies for Protein Complex Production and Characterization

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 896))

Abstract

Recombinant overexpression of a protein or a protein complex using any specific heterologous host can be an overwhelming challenge. The reasons may range from low yield and poor solubility of a single-subunit enzyme to the wrong stoichiometry or the incomplete assembly of a multiprotein complex. Whatever the reason, overcoming the difficulties will take the researcher into a journey through the seemingly countless options that exist for protein expression. While some choices stand to reason fairly straightforwardly (e.g., using Escherichia coli for the production of bacterial enzymes), most other choices do not need to be so self-revealing. Here, we attempt to portrait the canvas of available hosts for heterologous expression of many different protein classes and complexes and offer guidance as to which expression host may be more suitable to the problem at hand. The guidance in this chapter must be taken only as a rough indication which will have to be checked against the available literature and corroborated by experiment. It is not only expected but also welcome that, as more knowledge is gathered about the performance of hosts and protein types and new expression systems develop, the information in this chapter will have to be updated and refined.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 149.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 199.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 199.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Rosano GL, Ceccarelli EA (2014) Recombinant protein expression in Escherichia coli: advances and challenges. Front Microbiol 5:172

    PubMed  PubMed Central  Google Scholar 

  2. Strub MP, Hoh F, Sanchez JF, Strub JM, Bock A, Aumelas A, Dumas C (2003) Selenomethionine and selenocysteine double labeling strategy for crystallographic phasing. Structure 11(11):1359–1367

    Article  CAS  PubMed  Google Scholar 

  3. Cai M, Huang Y, Sakaguchi K, Clore GM, Gronenborn AM, Craigie R (1998) An efficient and cost-effective isotope labeling protocol for proteins expressed in Escherichia coli. J Biomol NMR 11(1):97–102

    Article  CAS  PubMed  Google Scholar 

  4. Hochkoeppler A (2013) Expanding the landscape of recombinant protein production in Escherichia coli. Biotechnol Lett 35(12):1971–1981

    Article  CAS  PubMed  Google Scholar 

  5. Bieniossek C, Nie Y, Frey D, Olieric N, Schaffitzel C, Collinson I, Romier C, Berger P, Richmond TJ, Steinmetz MO, Berger I (2009) Automated unrestricted multigene recombineering for multiprotein complex production. Nat Methods 6(6):447–450

    Article  CAS  PubMed  Google Scholar 

  6. Liebl W, Angelov A, Juergensen J, Chow J, Loeschcke A, Drepper T, Classen T, Pietruszka J, Ehrenreich A, Streit WR, Jaeger KE (2014) Alternative hosts for functional (meta)genome analysis. Appl Microbiol Biotechnol 98(19):8099–8109

    Article  CAS  PubMed  Google Scholar 

  7. Hwang PM, Pan JS, Sykes BD (2014) Targeted expression, purification, and cleavage of fusion proteins from inclusion bodies in Escherichia coli. FEBS Lett 588(2):247–252

    Article  CAS  PubMed  Google Scholar 

  8. Bill RM (2014) Playing catch-up with Escherichia coli: using yeast to increase success rates in recombinant protein production experiments. Front Microbiol 5:85

    Article  PubMed  PubMed Central  Google Scholar 

  9. Bieniossek C, Imasaki T, Takagi Y, Berger I (2012) MultiBac: expanding the research toolbox for multiprotein complexes. Trends Biochem Sci 37(2):49–57

    Article  CAS  PubMed  Google Scholar 

  10. Kriz A, Schmid K, Baumgartner N, Ziegler U, Berger I, Ballmer-Hofer K, Berger P (2010) A plasmid-based multigene expression system for mammalian cells. Nat Commun 1:120

    Article  PubMed  Google Scholar 

  11. An Y, Yumerefendi H, Mas PJ, Chesneau A, Hart DJ (2011) ORF-selector ESPRIT: a second generation library screen for soluble protein expression employing precise open reading frame selection. J Struct Biol 175(2):189–197

    Article  CAS  PubMed  Google Scholar 

  12. Yumerefendi H, Tarendeau F, Mas PJ, Hart DJ (2010) ESPRIT: an automated, library-based method for mapping and soluble expression of protein domains from challenging targets. J Struct Biol 172(1):66–74

    Article  CAS  PubMed  Google Scholar 

  13. Nie Y, Viola C, Bieniossek C, Trowitzsch S, Vijay-Achandran LS, Chaillet M, Garzoni F, Berger I (2009) Getting a grip on complexes. Curr Genomics 10(8):558–572

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Fernandez FJ, Vega MC (2013) Technologies to keep an eye on: alternative hosts for protein production in structural biology. Curr Opin Struct Biol 23(3):365–373

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was funded by the Spanish Ministry of Science and Innovation grant CTQ2015-66206-C2-2-R and the European Commission (Framework Programme 7) (FP7) project ComplexINC No. 279039 to M.C.V.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to M. Cristina Vega .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2016 Springer International Publishing Switzerland

About this chapter

Cite this chapter

Fernández, F.J., Vega, M.C. (2016). Choose a Suitable Expression Host: A Survey of Available Protein Production Platforms. In: Vega, M. (eds) Advanced Technologies for Protein Complex Production and Characterization. Advances in Experimental Medicine and Biology, vol 896. Springer, Cham. https://doi.org/10.1007/978-3-319-27216-0_2

Download citation

Publish with us

Policies and ethics