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Applied Microbiology and Biotechnology

, Volume 76, Issue 3, pp 633–642 | Cite as

Comparative analysis of twin-arginine (Tat)-dependent protein secretion of a heterologous model protein (GFP) in three different Gram-positive bacteria

  • Daniel Meissner
  • Angela Vollstedt
  • Jan Maarten van Dijl
  • Roland FreudlEmail author
Applied Genetics and Molecular Biotechnology

Abstract

In contrast to the general protein secretion (Sec) system, the twin-arginine translocation (Tat) export pathway allows the translocation of proteins across the bacterial plasma membrane in a fully folded conformation. Due to this feature, the Tat pathway provides an attractive alternative to the secretory production of heterologous proteins via the Sec system. In this study, the potential for Tat-dependent heterologous protein secretion was compared in the three Gram-positive bacteria Staphylococcus carnosus, Bacillus subtilis, and Corynebacterium glutamicum using green fluorescent protein (GFP) as a model protein. In all three microorganisms, fusion of a Tat signal peptide to GFP resulted in its Tat-dependent translocation across the corresponding cytoplasmic membranes. However, striking differences with respect to the final localization and folding status of the exported GFP were observed. In S. carnosus, GFP was trapped entirely in the cell wall and not released into the supernatant. In B. subtilis, GFP was secreted into the supernatant, however, in an inactive form. In contrast, C. glutamicum effectively secreted active GFP. Our results clearly demonstrate that a comparative evaluation of different Gram-positive host microorganisms is a crucial step on the way to an efficient Tat-mediated secretory production process for a desired heterologous target protein.

Keywords

Twin arginine translocation Protein secretion Green fluorescent protein Gram-positive 

Notes

Acknowledgments

We are very grateful to H. Sahm for his strong support of our work on bacterial protein secretion during now almost 20 years. We thank A. Bida for excellent technical assistance. R. Freudl and J.M. van Dijl were supported in part by European Union Grant LSHG-CT-2004-005257.

Supplementary material

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Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Daniel Meissner
    • 1
  • Angela Vollstedt
    • 1
  • Jan Maarten van Dijl
    • 2
  • Roland Freudl
    • 1
    Email author
  1. 1.Institut für Biotechnologie 1Forschungszentrum Jülich GmbHJülichGermany
  2. 2.Department of Medical MicrobiologyUniversity Medical Center Groningen and University of GroningenGroningenThe Netherlands

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