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Comparison of periplasmic and intracellular expression of Arabidopsis thionin proproteins in E. coli

Authors
  • Abbas, Amjad1
  • Plattner, Stephan1
  • Shah, Kausar Hussain1
  • Bohlmann, Holger1
  • 1 University of Natural Resources and Life Sciences, Vienna, Division of Plant Protection, Department of Crop Sciences, UFT Tulln, Konrad Lorenz Str. 24, Tulln, 3430, Austria , Tulln (Austria)
Type
Published Article
Journal
Biotechnology Letters
Publisher
Springer-Verlag
Publication Date
Mar 21, 2013
Volume
35
Issue
7
Pages
1085–1091
Identifiers
DOI: 10.1007/s10529-013-1180-z
Source
Springer Nature
Keywords
License
Green

Abstract

Thionins are antimicrobial plant peptides produced as preproproteins consisting of a signal peptide, the thionin domain, and a so-called acidic domain. Only thionin itself has been isolated from plants. To study the processing of the precursor, it has to be produced in a heterologous system. Since both domains contain several cysteines and, due to the known antimicrobial activity of the thionin, we tested the expression of all four Arabidopsis proproteins as fusion proteins. Periplasmic expression as fusion with maltose binding protein was not successful but cytoplasmic expression as His-tagged TRX fusion proteins with a TEV recognition sequence resulted in proteins of correct size. Use of the SHuffle strain C3030 further improved the expression. Fusion proteins inhibited growth of Escherichia coli. They could be cleaved by TEV protease, releasing authentic proproteins without any additional amino acid at the N-terminus.

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