Applied Microbiology and Biotechnology

, Volume 78, Issue 1, pp 139–145 | Cite as

Changing the N-terminal sequence protects recombinant Plasmodium falciparum circumsporozoite protein from degradation in Pichia pastoris

  • Qingfeng Zhang
  • Feng Ding
  • Xiangyang Xue
  • Xindong Xu
  • Weiqing Pan
Applied genetics and molecular biotechnology
First Online:
Received:
Revised:
Accepted:

Abstract

Proteolytic degradation is the primary obstacle in the use of the yeast Pichia pastoris for the expression of recombinant proteins. During the production of a recombinant Plasmodium falciparum circumsporozoite protein in this system, the (NANP) n repeats region at the N-terminus were completely proteolytically degraded. To remove the potential proteolytic site within the recombinant protein, different strategies were tried, including adjusting the cultivation conditions and mutating the sequence at the junction of the repeat domain and C-terminal region, but the degradation continued. However, modification of the N-terminal sequence by adding an epitope-based peptide to the N-terminus not only protected the repeat domain from cleavage by native proteases during longer induction in the yeast host and purification process, but also stabilized this recombinant protein emulsified with adjuvant ISA720 for at least 6 months. The results showed that proteolytic degradation of the recombinant circumsporozoite protein produced in P. pastoris was amino acid sequence (NANP)-specific, and that this effect was likely dependent on the conformation of the recombinant protein.

Keywords

Pichia pastoris Recombinant protein Proteolytic degradation Plasmodium falciparum Circumsporozoite protein 
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Notes

Acknowledgements

This work was supported by a grant from the National Basic Research Program (973 program) (no. 2007CB513100), the National Natural Science Foundation of China (no. 30430610) and a grant from the UNDP/World Bank/WHO Special Program for Research and Training in Tropical Diseases (TDR).

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Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Qingfeng Zhang
    • 1
  • Feng Ding
    • 1
  • Xiangyang Xue
    • 1
  • Xindong Xu
    • 1
  • Weiqing Pan
    • 1
  1. 1.Department of Pathogen Biology and State Key Laboratory of Medical ImmunologySecond Military Medical UniversityShanghaiChina

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