NDLI: Process optimization for increased yield of surface-expressed protein in Escherichia coli

Content Provider	Springer Nature Link
Author	Jarmander, Johan Jaschek, Lars Lundh, Susanna Larsson, Gen Gustavsson, Martin
Copyright Year	2014
Abstract	The autotransporter family of Gram-negative protein exporters has been exploited for surface expression of recombinant passenger proteins. While the passenger in some cases was successfully translocated, a major problem has been low levels of full-length protein on the surface due to proteolysis following export over the cytoplasmic membrane. The aim of the present study was to increase the surface expression yield of the model protein SefA, a Salmonella enterica fimbrial subunit with potential for use in vaccine applications, by reducing this proteolysis through process design using Design of Experiments methodology. Cultivation temperature and pH, hypothesized to influence periplasmic protease activity, as well as inducer concentration were the parameters selected for optimization. Through modification of these parameters, the total surface expression yield of SefA was increased by 200 %. At the same time, the yield of full-length protein was increased by 300 %, indicating a 33 % reduction in proteolysis.
Starting Page	1685
Ending Page	1693
Page Count	9
File Format	PDF
ISSN	16157591
Journal	Bioprocess Engineering
Volume Number	37
Issue Number	8
e-ISSN	16157605
Language	English
Publisher	Springer Berlin Heidelberg
Publisher Date	2014-02-14
Publisher Place	Berlin, Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Surface expression Autotransport Process optimization Proteolysis Live vaccines Biotechnology Industrial and Production Engineering Environmental Engineering/Biotechnology Industrial Chemistry/Chemical Engineering Food Science
Content Type	Text
Resource Type	Article
Subject	Medicine Bioengineering Biotechnology

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1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
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4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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