NDLI: A dual expression platform to optimize the soluble production of heterologous proteins in the periplasm of Escherichia coli

Content Provider	Springer Nature Link
Author	Kraft, Mario Knüpfer, Uwe Wenderoth, Rolf Kacholdt, André Pietschmann, Patricia Hock, Björn Horn, Uwe
Copyright Year	2007
Abstract	The functional analysis of individual proteins or of multiprotein complexes—since the completion of several genome sequencing projects—is in focus of current scientific work. Many heterologous proteins contain disulfide-bonds, required for their correct folding and activity, and therefore, need to be transported to the periplasm. The production of soluble and functional protein in the periplasm often needs target-specific regulatory genetic elements, leader peptides, and folding regimes. Usually, the optimization of periplasmic expression is a step-wise and time-consuming procedure. To overcome this problem we developed a dual expression system, containing a degP-promoter-based reporter system and a highly versatile plasmid set. This combines the differential protein expression with the selection of a target-specific expression plasmid. For the validation of this expression tool, two different molecular formats of a recombinant antibody directed to the human epidermal growth factor receptor and human 11β-hydroxysteroid dehydrogenase type 2 (11β-HSD2) were used. By application of this expression system we demonstrated that the amount of functional protein is inversely proportional to the on-line luciferase signal. We showed that this technology offers a simple tool to evaluate and improve the yield of functionally expressed proteins in the periplasm, which depends on the used regulatory elements and folding strategies.
Starting Page	1413
Ending Page	1422
Page Count	10
File Format	PDF
ISSN	01757598
Journal	Applied Microbiology and Biotechnology
Volume Number	76
Issue Number	6
e-ISSN	14320614
Language	English
Publisher	Springer-Verlag
Publisher Date	2007-08-08
Publisher Place	Berlin, Heidelberg
Access Restriction	Subscribed
Subject Keyword	Microbial Genetics and Genomics Microbiology Biotechnology
Content Type	Text
Resource Type	Article
Subject	Medicine Applied Microbiology and Biotechnology Biotechnology

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RETRACTED ARTICLE: Strategies for efficient production of heterologous proteins in Escherichia coli

Retraction Note: Strategies for efficient production of heterologous proteins in Escherichia coli

Effect of folding factors in rescuing unstable heterologous lipase B to enhance its overexpression in the periplasm of Escherichia coli

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RETRACTED ARTICLE: Strategies for efficient production of heterologous proteins in Escherichia coli

Retraction Note: Strategies for efficient production of heterologous proteins in Escherichia coli

Effect of folding factors in rescuing unstable heterologous lipase B to enhance its overexpression in the periplasm of Escherichia coli

Improving heterologous polyketide production in Escherichia coli by transporter engineering

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