Dissection of an old protein reveals a novel application: domain D of Staphylococcus aureus Protein A (sSpAD) as a secretion - tag

Microbial Cell Factories

Research

Rainer Schneider¹  Bernhard Auer¹  Thomas Heel¹  Michael Paal¹ 

[1] Austrian Center of Industrial Biotechnology (ACIB), TU Graz, Petersgasse 14, A-801, Graz, Austria;Institute of Biochemistry, University of Innsbruck, Peter-Mayr-Strasse 1a, A-6020, Innsbruck, Austria;Center for Molecular Biosciences (CMBI), University of Innsbruck, Peter-Mayr-Strasse 1a, A-6020, Innsbruck, Austria;

关键词: Shake Flask Cultivation;    Maltose Binding Protein;    Secretion Capacity;    Recombinant Fusion Protein;    Human Superoxide Dismutase;   

DOI  :  10.1186/1475-2859-9-92

 received in 2010-08-26, accepted in 2010-11-23,  发布年份 2010

来源: Springer

【 摘 要 】

BackgroundEscherichia coli as a frequently utilized host organism for recombinant protein production offers different cellular locations with distinct qualities. The periplasmic space is often favored for the production of complex proteins due to enhanced disulfide bond formation, increased target product stability and simplified downstream processing. To direct proteins to the periplasmic space rather small proteinaceus tags that can be used for affinity purification would be advantageous.ResultsWe discovered that domain D of the Staphylococcus aureus protein A was sufficient for the secretion of various target proteins into the periplasmic space of E. coli. Our experiments indicated the Sec pathway as the mode of secretion, although N-terminal processing was not observed. Furthermore, the solubility of recombinant fusion proteins was improved for proteins prone to aggregation.The tag allowed a straightforward affinity purification of recombinant fusion protein via an IgG column, which was exemplified for the target protein human superoxide dismutase 1 (SOD).ConclusionsIn this work we present a new secretion tag that combines several advantages for the production of recombinant proteins in E. coli. Domain D of S. aureus protein A protects the protein of interest against N-terminal degradation, increases target protein solubility and enables a straight-forward purification of the recombinant protein using of IgG columns.

【 授权许可】

Unknown   
© Heel et al; licensee BioMed Central Ltd. 2010. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.