Skip to main content
Log in

pHsh vectors, a novel expression system of Escherichia coli for the large-scale production of recombinant enzymes

  • Original Research Paper
  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

Gene expression system Hsh is developed to increase enzyme production and to decrease the cost in the induction of gene expression in Escherichia coli. The vectors of Hsh system were constructed by combining a synthesized heat-shock promoter with a synthesized terminator and an origin of replication derived from pUC19 in which the expression of foreign genes was regulated by an alternative sigma factor, σ32 of E. coli. In comparison, the Hsh promoter gave a 2.4-fold higher production for xynIII gene encoding a xylanase than existing heat-shock inducible promoter p L, 1.2-fold and 3-fold production for xar gene encoding a arabinosidase than trc and T7 promoter, respectively. The flow-in-heat technique created a rapid rise in temperature for effective induction of gene expression in bioreactor scale.

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

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Amann E, Brosius J, Ptashne M (1983) Vectors bearing a hybrid trp-lac promoter useful for regulated expression of cloned genes in Escherichia coli. Gene 25:167–178

    Article  CAS  PubMed  Google Scholar 

  • Arsene F, Tomoyasu T, Bukau B (2000) The heat-shock response of Escherichia coli. Int J Food Microbiol 55:3–9

    Article  CAS  PubMed  Google Scholar 

  • Bernard HU, Helinski DR (1979) Use of the lambda phage promoter P L to promote gene expression in hybrid plasmid cloning vehicles. Methods Enzymol 68:482–492

    Article  CAS  PubMed  Google Scholar 

  • Binz T, Canevascini G (1996) Xylanases from the Dutch elm disease pathogens Ophiostoma ulmi and Ophiostoma novo-ulmi. Physiol Mol Plant Pathol 49:159–175

    Article  CAS  Google Scholar 

  • Calendar R, Erickson JW, Halling C, Nolte A (1988) Deletion and insertion mutations in the rpoH gene of Escherichia coli that produce functional sigma 32. J Bacteriol 150:815–825

    Google Scholar 

  • Cowing DW, Bardwell JC, Craig EA, Woolford C, Hendrix RW, Gross CA (1985) Consensus sequence for Escherichia coli heat-shock gene promoters. Proc Natl Acad Sci USA 82:2679–2683

    Article  CAS  PubMed  Google Scholar 

  • Gold L (1990) Expression of heterologous proteins in Escherichia coli. Methods Enzymol 185:11–14

    Article  CAS  PubMed  Google Scholar 

  • Miller JH (1992) A short course in bacterial genetics (Handbook). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA

    Google Scholar 

  • Remaut E, Stanssens P, Fiers W (1981) Plasmid vectors for high-efficiency expression controlled by the P L promoter of coliphage lambda. Gene 15:81–93

    Article  CAS  PubMed  Google Scholar 

  • Shao W, Pei J, Li X (2005) The method for over-expression of the xylanase B from Thermotoga maritima in Escherichia coli. China Patent, ZL 03131524.0

  • Studier FW, Moffatt BA (1986) Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol 189:113–130

    Article  CAS  PubMed  Google Scholar 

  • Tilly K, Spence J, Georgopoulos C (1989) Modulation of stability of the Escherichia coli heat-shock regulatory factor sigma. J Bacteriol 171:1585–1589

    CAS  PubMed  Google Scholar 

  • Wu H, Pei J, Wu G, Shao W (2008) Overexpression of GH10 endoxylanase XynB from T. maritima in E. coli by a novel vector with potential for industrial application. Enzyme Microb Technol 42:230–234

    Article  CAS  Google Scholar 

  • Xue Y, Wu A, Zeng H, Shao W (2006) High-level expression of an α-l-arabinofuranosidase from Thermotoga maritima in Escherichia coli for the production of xylobiose from xylan. Biotechnol Lett 28:351–356

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from the sub-project of ‘973’ in China (2004CB719600), and National Natural Science Foundation of China (30770061).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Weilan Shao.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material (DOCX 17 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wu, H., Pei, J., Jiang, Y. et al. pHsh vectors, a novel expression system of Escherichia coli for the large-scale production of recombinant enzymes. Biotechnol Lett 32, 795–801 (2010). https://doi.org/10.1007/s10529-010-0223-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10529-010-0223-y

Keywords

Navigation