Skip to main content

A Generic Protocol for Intracellular Expression of Recombinant Proteins in Bacillus subtilis

  • Protocol
  • First Online:
Book cover Heterologous Gene Expression in E.coli

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1586))

Abstract

Bacillus subtilis (B. subtilis) is a potential and attractive host for the production of recombinant proteins. Different expression systems for B. subtilis have been developed recently, and various target proteins have been recombinantly synthesized and purified using this host. In this chapter, we introduce a generic protocol to express a recombinant protein in B. subtilis. It includes protocols for (1) using our typical expression vector (plasmid pHT254) to introduce a target gene, (2) transformation of the target vector into B. subtilis, and (3) evaluation of the actual expression of a recombinant protein.

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

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 109.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 219.99
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. Adrio JL, Demain AL (2010) Recombinant organisms for production of industrial products. Bioeng Bugs 1:116–131

    Article  PubMed  Google Scholar 

  2. Demain AL, Vaishnav P (2009) Production of recombinant proteins by microbes and higher organisms. Biotechnol Adv 27:297–306

    Article  CAS  PubMed  Google Scholar 

  3. Sahdev S, Khattar SK, Saini KS (2008) Production of active eukaryotic proteins through bacterial expression systems: a review of the existing biotechnology strategies. Mol Cell Biochem 307:249–264

    Article  CAS  PubMed  Google Scholar 

  4. Schallmey M, Singh A, Ward OP (2004) Developments in the use of Bacillus species for industrial production. Can J Microbiol 50:1–17

    Article  CAS  PubMed  Google Scholar 

  5. Schumann W (2007) Production of recombinant proteins in Bacillus subtilis. Adv Appl Microbiol 62:137–189

    Article  CAS  PubMed  Google Scholar 

  6. Tjalsma H, Antelmann H, Jongbloed JDH et al (2004) Proteomics of protein secretion by Bacillus subtilis: separating the “secrets” of the secretome. Microbiol Mol Biol Rev 68:207–233

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Nguyen HD, Nguyen QA, Ferreira RC et al (2005) Construction of plasmid-based expression vectors for Bacillus subtilis exhibiting full structural stability. Plasmid 54:241–248

    Article  CAS  PubMed  Google Scholar 

  8. Phan TTP, Nguyen HD, Schumann W (2006) Novel plasmid-based expression vectors for intra- and extracellular production of recombinant proteins in Bacillus subtilis. Protein Expr Purif 46:189–195

    Article  CAS  PubMed  Google Scholar 

  9. Nguyen HD, Phan TTP, Schumann W (2007) Expression vectors for the rapid purification of recombinant proteins in Bacillus subtilis. Curr Microbiol 55:89–93

    Article  CAS  PubMed  Google Scholar 

  10. Phan TTP, Nguyen HD, Schumann W (2012) Development of a strong intracellular expression system for Bacillus subtilis by optimizing promoter elements. J Biotechnol 157:167–172

    Article  CAS  PubMed  Google Scholar 

  11. Phan TTP, Nguyen HD, Schumann W (2013) Construction of a 5′-controllable stabilizing element (CoSE) for over-production of heterologous proteins at high levels in Bacillus subtilis. J Biotechnol 168:32–39

    Article  CAS  PubMed  Google Scholar 

  12. Phan TTP, Tran LT, Schumann W et al (2015) Development of Pgrac100-based expression vectors allowing high protein production levels in Bacillus subtilis and relatively low basal expression in Escherichia coli. Microb Cell Fact 14:72

    Article  PubMed  PubMed Central  Google Scholar 

  13. Hirata H, Fukazawa T, Negoro S et al (1986) Structure of a beta-galactosidase gene of Bacillus stearothermophilus. J Bacteriol 166:722–727

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Saito H, Shibata T, Ando T (1979) Mapping of genes determining nonpermissiveness and host-specific restriction to bacteriophages in Bacillus subtilis Marburg. Mol Gen Genet 170:117–122

    Article  CAS  PubMed  Google Scholar 

  15. Inoue H, Nojima H, Okayama H (1990) High efficiency transformation of Escherichia coli with plasmids. Gene 96:23–28

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This research was funded by the Vietnam National Foundation for Science and Technology Development (Nafosted) under grant number 106-NN.02-2015.24. The research was also supported by TWAS under research grants 14-201 RG/BIO/AS_G.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Hoang Nguyen .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2017 Springer Science+Business Media LLC

About this protocol

Cite this protocol

Phan, T., Huynh, P., Truong, T., Nguyen, H. (2017). A Generic Protocol for Intracellular Expression of Recombinant Proteins in Bacillus subtilis . In: Burgess-Brown, N. (eds) Heterologous Gene Expression in E.coli. Methods in Molecular Biology, vol 1586. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6887-9_21

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-6887-9_21

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6885-5

  • Online ISBN: 978-1-4939-6887-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics