Expression and purification of a nanostructure-forming peptide

Hartmann, B., Kaar, W., Falconer, R., Zeng, B. and Middelberg, A.P.J. (2008) Expression and purification of a nanostructure-forming peptide. Journal of Biotechnology, 135 1: 85-91. doi:10.1016/j.jbiotec.2008.03.003


Author Hartmann, B.
Kaar, W.
Falconer, R.
Zeng, B.
Middelberg, A.P.J.
Title Expression and purification of a nanostructure-forming peptide
Journal name Journal of Biotechnology   Check publisher's open access policy
ISSN 0168-1656
1873-4863
Publication date 2008-05-20
Year available 2008
Sub-type Article (original research)
DOI 10.1016/j.jbiotec.2008.03.003
Volume 135
Issue 1
Start page 85
End page 91
Total pages 7
Editor A. Puehler
Place of publication Amsterdam, Netherlands
Publisher Elsevier Science
Collection year 2009
Language eng
Subject C1
860899 Human Pharmaceutical Products not elsewhere classified
100302 Bioprocessing, Bioproduction and Bioproducts
100799 Nanotechnology not elsewhere classified
Abstract Peptides have recently attracted interest as building blocks for the assembly of novel functional materials including switchable surfactants, nanocoatings, hydrogels and aqueous vesicles. We expressed a betasheet forming peptide that has been widely studied in self-assembly processing, P-11-2, as a monomer, dimer, tetramer and nonamer fused to an insoluble expression partner, ketosteroid isomerase, using minimal media. Expression was followed by whole cell extraction and isolation of the fusion protein to greater than 90% purity via a single immobilised metal affinity chromatography (IMAC) step. Peptides were chemically cleaved from each other and from the fusion partner, followed by acetone precipitation of the contaminating protein fragments. Pure peptide was recovered by reversed-phase HPLC. The expression level of the fusion protein decreased as the peptide concatamer number increased, as did the efficiency of the chemical cleavage, making the single-peptide process the most efficient overall. Applying this laboratory process to the single-peptide fusion protein nevertheless resulted in a pure peptide yield of greater than 30% of the expressed peptide. Crown Copyright (C) 2008 Published by Elsevier B.V. All rights reserved.
Keyword Peptide production
Concatamer
Chemical cleavage
Inclusion body
Expression
Q-Index Code C1
Q-Index Status Confirmed Code

 
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Created: Thu, 16 Apr 2009, 15:02:10 EST by Amanda Lee on behalf of Aust Institute for Bioengineering & Nanotechnology