Functional cell surface display and controlled secretion of diverse Agarolytic enzymes by Escherichia coli with a novel ligation-independent cloning vector based on the autotransporter YfaL.

Applied and Environmental Microbiology
Hyeok-Jin KoIn-Geol Choi

Abstract

Autotransporters have been employed as the anchoring scaffold for cell surface display by replacing their passenger domains with heterologous proteins to be displayed. We adopted an autotransporter (YfaL) of Escherichia coli for the cell surface display system. The critical regions in YfaL for surface display were identified for the construction of a ligation-independent cloning (LIC)-based display system. The designed system showed no detrimental effect on either the growth of the host cell or overexpressing heterologous proteins on the cell surface. We functionally displayed monomeric red fluorescent protein (mRFP1) as a reporter protein and diverse agarolytic enzymes from Saccharophagus degradans 2-40, including Aga86C and Aga86E, which previously had failed to be functional expressed. The system could display different sizes of proteins ranging from 25.3 to 143 kDa. We also attempted controlled release of the displayed proteins by incorporating a tobacco etch virus protease cleavage site into the C termini of the displayed proteins. The maximum level of the displayed protein was 6.1 × 10(4) molecules per a single cell, which corresponds to 5.6% of the entire cell surface of actively growing E. coli.

References

Oct 5, 1990·Journal of Molecular Biology·S F AltschulD J Lipman
Oct 25, 1990·Nucleic Acids Research·C Aslanidis, P J de Jong
Apr 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·C T ChungR H Miller
Jun 24, 1970·Journal of Chromatography·M Duckworth, W Yaphe
Dec 26, 2001·Nucleic Acids Research·Alex BatemanErik L L Sonnhammer
Jun 1, 2002·Journal of Biotechnology·Patrik SamuelsonStefan Ståhl
Jun 13, 2002·Proceedings of the National Academy of Sciences of the United States of America·Robert E CampbellRoger Y Tsien
Jun 30, 2004·Journal of Molecular Biology·Jannick Dyrløv BendtsenSøren Brunak
Oct 2, 2004·Biotechnology and Bioengineering·Zhizhuang XiaoAdrian Tsang
Mar 9, 2006·Protein Science : a Publication of the Protein Society·Paola MaraniGunnar von Heijne
May 5, 2006·Applied and Environmental Microbiology·Nathan A EkborgSteven W Hutcheson
Jun 22, 2006·Microbial Cell Factories·Nancy Rutherford, Michael Mourez
Nov 14, 2006·Nucleic Acids Research·Derek WilsonJulian Gough
Mar 6, 2007·Applied and Environmental Microbiology·Wei-wei Zhang, Li Sun
May 18, 2007·Annual Review of Microbiology·Nathalie Dautin, Harris D Bernstein
Aug 31, 2007·Current Opinion in Structural Biology·Patrick S Daugherty
Nov 13, 2007·Nature Structural & Molecular Biology·Travis J BarnardSusan K Buchanan
Oct 8, 2008·Nucleic Acids Research·Brandi L CantarelBernard Henrissat
Oct 24, 2009·Proceedings of the National Academy of Sciences of the United States of America·Raffaele Ieva, Harris D Bernstein
Nov 19, 2009·Nucleic Acids Research·Robert D FinnAlex Bateman
May 8, 2010·Microbiology·Timothy J WellsMark A Schembri
Aug 31, 2010·Journal of Bacteriology·Elvira MarínLuis Ángel Fernández
Nov 27, 2010·Nucleic Acids Research·Aron Marchler-BauerStephen H Bryant
Aug 4, 2011·Biochemical and Biophysical Research Communications·Sung Chul HaIn-Geol Choi

❮ Previous
Next ❯

Citations

Aug 12, 2014·Applied Microbiology and Biotechnology·Jan SchüürmannJoachim Jose
Jul 17, 2013·Critical Reviews in Microbiology·Toon NicolayStijn Spaepen
Jun 14, 2018·Journal of Agricultural and Food Chemistry·Duleepa PathirajaIn-Geol Choi

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.