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Improved detection ...
Improved detection and performance of surface expression from the AIDA-I autotransporter
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- Jarmander, Johan, 1983- (author)
- KTH,Bioprocessteknik
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- Larsson, Gen, Professor (thesis advisor)
- KTH,Bioprocessteknik
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- Rova, Ulrika, Professor, bitr (opponent)
- Luleå Tekniska Universitet
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- Ezcurra, Ines, Universitetslektor (opponent)
- KTH,Glykovetenskap
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(creator_code:org_t)
- ISBN 9789175017082
- Stockholm : KTH Royal Institute of Technology, 2013
- English 53 s.
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Series: TRITA-BIO-Report, 1654-2312 ; 2013:7
- Related links:
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https://kth.diva-por... (primary) (Raw object)
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https://urn.kb.se/re...
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Abstract
Subject headings
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- 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.
Subject headings
- TEKNIK OCH TEKNOLOGIER -- Industriell bioteknik -- Bioprocessteknik (hsv//swe)
- ENGINEERING AND TECHNOLOGY -- Industrial Biotechnology -- Bioprocess Technology (hsv//eng)
Keyword
- surface expression
- autotransport
- AIDA-I
- Escherichia coli
- proteolysis
- detection tag
Publication and Content Type
- vet (subject category)
- lic (subject category)
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