Enhanced translocation of recombinant proteins via the Tat pathway with chaperones in Escherichia coli

Ya Fang Lee, Hsin Yi Hsieh, Danielle Tullman-Ercek, Tang Kang Chiang, Raymond J. Turner, Sung Chyr Lin*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

The twin-arginine translocation (Tat) pathway is capable of translocating folded proteins into the periplasm of Gram-negative bacteria and thus holds great potential for the expression of recombinant proteins in Escherichia coli. Nevertheless, this promise has been hampered by the low translocation efficiency. In this study, we demonstrate that the co-expression of DmsD, a system specific cytoplasmic chaperone similar to TorD, in conjunction with the DmsA signal peptide can enhance the translocation of the GFP fusion protein by 28.2%. We further show the presence of cross-activity between DmsD and TorD for the DmsA and TorA leader-fusions. The co-expression of DmsD and TorD enhances the translocation of ssTorA-GFP fusion and ssDmsA-GFP fusion by 28.6% and 46.6%, respectively. It was also observed that the co-expression of DmsD led to a reduction in the formation of GFP inclusion bodies, whereas the co-expression of TorD primarily led to a reduction in proteolysis by the Clp system. It is concluded that DmsD and TorD enhance protein translocation via the Tat pathway by providing activity against protein aggregation and/or proteolysis.

Original languageEnglish (US)
Pages (from-to)540-546
Number of pages7
JournalJournal of the Taiwan Institute of Chemical Engineers
Volume41
Issue number5
DOIs
StatePublished - Sep 2010

Funding

We are grateful to Drs F. Sargent and G. Georgiou for the precious plasmids and antiserum used in this study. This work is supported by a grant, NSC95-2214-E-005-029 , from the National Science Council, Taiwan . RJT is supported by CIHR. A grant to Dr. SC Lin from the ATU plan, Ministry of Education, Taiwan is appreciated.

Keywords

  • DmsD
  • Green fluorescence protein
  • Secretion
  • TorD
  • Twin-arginine pathway

ASJC Scopus subject areas

  • General Chemistry
  • General Chemical Engineering

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