Comparative analysis of twin-arginine (Tat)-dependent protein secretion of a heterologous model protein (GFP) in three different Gram-positive bacteria |
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Authors: | Daniel Meissner Angela Vollstedt Jan Maarten van Dijl Roland Freudl |
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Institution: | (1) Institut für Biotechnologie 1, Forschungszentrum Jülich GmbH, 52425 Jülich, Germany;(2) Department of Medical Microbiology, University Medical Center Groningen and University of Groningen, Hanzeplein 1, P.O.Box 30001, 9700 RB Groningen, The Netherlands |
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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.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.
This paper is dedicated to Hermann Sahm on the occasion of his 65th birthday. |
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Keywords: | Twin arginine translocation Protein secretion Green fluorescent protein Gram-positive |
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