Improved detection and performance of surface expression from the AIDA-I autotransporter
2013 (English)Licentiate thesis, comprehensive summary (Other academic)
Surface expression of recombinant proteins has attracted a lot of attention due to its potential in applications such as enzyme production, vaccine delivery and bioremediation. Autotransporters have been used for surface expression of a variety of proteins, but the expression systems reported in literature have typically been inflexible and incapable of detecting proteolysis, thereby limiting surface expression yield.
In this thesis, a modular surface expression system, utilizing dual tag detection, was therefore created. It was based on the adhesin involved in diffuse adherence (AIDA-I) autotransporter, and was here used to express the model proteins SefA and H:gm on the cell surface of Escherichia coli. Due to the dual tag detection system, proteolysed H:gm could be successfully verified on the cell surface. By optimizing cultivation conditions, surface expression yield of SefA was increased by 300 %, and proteolysis reduced by 33 %. While proteolysis could not be eliminated completely, the work presented in this thesis is a major step towards a general system for surface expression of a wide range of proteins in varied applications.
Place, publisher, year, edition, pages
Stockholm: KTH Royal Institute of Technology, 2013. , 53 p.
TRITA-BIO-Report, ISSN 1654-2312 ; 2013:7
surface expression, autotransport, AIDA-I, Escherichia coli, proteolysis, detection tag
IdentifiersURN: urn:nbn:se:kth:diva-121561ISBN: 978-91-7501-708-2OAI: oai:DiVA.org:kth-121561DiVA: diva2:619110
2013-05-31, AlbaNova, sal FB55, Roslagstullsbacken 21, Stockholm, 10:00 (English)
Rova, Ulrika, Professor, bitrEzcurra, Ines, Universitetslektor
Larsson, Gen, Professor
QC 201305062013-05-062013-05-022014-12-11Bibliographically approved
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