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Applied Microbiology and Biotechnology

, Volume 91, Issue 1, pp 133–141 | Cite as

Hydrophobin signal sequence mediates efficient secretion of recombinant proteins in Pichia pastoris

  • Kirsten Kottmeier
  • Kai Ostermann
  • Thomas Bley
  • Gerhard Rödel
Applied Genetics and Molecular Biotechnology

Abstract

Pichia pastoris is an important eukaryotic organism for the expression, processing, and secretion of recombinant proteins. Here, the secretion of enhanced green fluorescent protein (EGFP) in P. pastoris by using three novel secretion signals originating from the HFBI and HFBII class 2 hydrophobins of Trichoderma reesei was investigated. EGFP was fused to the carboxyl terminus of hydrophobin secretion signals and expressed under the control of the constitutive GAP promoter. In every case, recombinant EGFP entered the secretory pathway of P. pastoris. SDS-polyacrylamide gel electrophoresis, Western blot analysis of the cells' supernatant, and fluorescence measurements on single-cell level via flow cytometry confirmed the efficient secretion of EGFP mediated by the novel secretion sequences. In conclusion, the data clearly show that the secretion sequences derived from HFBI and HFBII of T. reesei have the potential to achieve an efficient secretion of heterologous proteins in P. pastoris. Due to the small size of the hydrophobin-derived secretion signals, their coding sequence can be easily introduced to the gene of interest by PCR.

Keywords

Secretion Hydrophobin Pichia pastoris Signal sequence Recombinant proteins EGFP 

Notes

Acknowledgment

We gratefully thank K. Zarschler for valuable discussion and critical reading of the manuscript.

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Kirsten Kottmeier
    • 1
    • 2
  • Kai Ostermann
    • 1
  • Thomas Bley
    • 2
  • Gerhard Rödel
    • 1
  1. 1.Institute of GeneticsDresden University of TechnologyDresdenGermany
  2. 2.Institute of Food Technology and Bioprocess EngineeringDresden University of TechnologyDresdenGermany

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