scholarly journals 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

2012 ◽  
Vol 78 (9) ◽  
pp. 3051-3058 ◽  
Author(s):  
Hyeok-Jin Ko ◽  
Eunhye Park ◽  
Joseph Song ◽  
Taek Ho Yang ◽  
Hee Jong Lee ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Cell Surface ◽  
Fluorescent Protein ◽  
Surface Display ◽  
Cell Surface Display ◽  
Heterologous Proteins ◽  
Display System ◽  
Reporter Protein ◽  
Content Type ◽  
Ligation Independent Cloning

ABSTRACTAutotransporters 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) ofEscherichia colifor 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 fromSaccharophagus degradans2-40, 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 × 104molecules per a single cell, which corresponds to 5.6% of the entire cell surface of actively growingE. coli.

scholarly journals Combinatorial cell surface display system in Escherichia coli for noninvasive colorectal cancer detection

2019 ◽  
Vol 2 (4) ◽  
pp. 185-196
Author(s):  
Chiayi Hsiao ◽  
Qingwei Fang ◽  
Shuyu Zheng ◽  
Peixiang He ◽  
Kairan Zhang ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Escherichia Coli ◽  
Cell Surface ◽  
Cancer Detection ◽  
Surface Display ◽  
Cell Surface Display ◽  
Display System ◽  
Cell Surface Display System

scholarly journals Enhanced Display of Lipase on the Escherichia coli Cell Surface, Based on Transcriptome Analysis

2009 ◽  
Vol 76 (3) ◽  
pp. 971-973 ◽  
Author(s):  
Jong Hwan Baek ◽  
Mee-Jung Han ◽  
Seung Hwan Lee ◽  
Sang Yup Lee
Keyword(s):  
Escherichia Coli ◽  
Cell Surface ◽  
Transcriptome Analysis ◽  
Surface Display ◽  
Cell Surface Display ◽  
Display System ◽  
E Coli ◽  
A Cell ◽  
Anchoring Motif ◽  
Cell Surface Display System

ABSTRACT A cell surface display system was developed using Escherichia coli OmpC as an anchoring motif. The fused Pseudomonas fluorescens SIK W1 lipase was successfully displayed on the surface of E. coli cells, and the lipase activity could be enhanced by the coexpression of the gadBC genes identified by transcriptome analysis.

scholarly journals Cell Surface Display of Poliovirus Receptor on Escherichia coli, a Novel Method for Concentrating Viral Particles in Water

2011 ◽  
Vol 77 (15) ◽  
pp. 5141-5148 ◽  
Author(s):  
Morteza Abbaszadegan ◽  
Absar Alum ◽  
Hamed Abbaszadegan ◽  
Valerie Stout
Keyword(s):  
Escherichia Coli ◽  
Cell Surface ◽  
Water Samples ◽  
Stop Codon ◽  
Surface Display ◽  
Cell Surface Display ◽  
Bacterial Cells ◽  
Content Type ◽  
E Coli ◽  
Poliovirus Receptor

ABSTRACTThe lack of efficient methods for concentrating viruses in water samples leads to underreporting of viral contamination in source water. A novel strategy for viral concentration was developed using the expression of target virus receptors on bacterial cells. Poliovirus type 1, the most studied enterovirus, was used as a surrogate for enteric viruses. The human poliovirus receptor (hPVR) gene was expressed on the surface ofEscherichia colicells by using the ice nucleation protein (INP) gene. ThehPVRgene was ligated to the 3′ end of theINPgene after the removal of the stop codon. The resulting open reading frame (ORF) was used for the projection of hPVR onto the outer membrane ofE. coli. Gene expression was tested by SDS-PAGE, Western blot, and dot blot analyses, and virion capture ability was confirmed by transmission electron microscopy. The application of engineeredE. colicells for capturing viruses in 1-liter samples of source and drinking water resulted in 75 to 99% procedural recovery efficiency. Cell surface display of viral receptors on bacterial cells opens a new prospect for an efficient and inexpensive alternative tool for capturing and concentrating waterborne viruses in water samples.

scholarly journals Preparation of Sticky Escherichia coli through Surface Display of an Adhesive Catecholamine Moiety

2013 ◽  
Vol 80 (1) ◽  
pp. 43-53 ◽  
Author(s):  
Joseph P. Park ◽  
Min-Jung Choi ◽  
Se Hun Kim ◽  
Seung Hwan Lee ◽  
Haeshin Lee
Keyword(s):  
Escherichia Coli ◽  
Cell Surface ◽  
Communication Systems ◽  
Cell Attachment ◽  
Surface Display ◽  
Content Type ◽  
E Coli ◽  
Material Surfaces ◽  
Mussel Adhesive ◽  
Adhesive Proteins

ABSTRACTMussels attach to virtually all types of inorganic and organic surfaces in aqueous environments, and catecholamines composed of 3,4-dihydroxy-l-phenylalanine (DOPA), lysine, and histidine in mussel adhesive proteins play a key role in the robust adhesion. DOPA is an unusual catecholic amino acid, and its side chain is called catechol. In this study, we displayed the adhesive moiety of DOPA-histidine onEscherichia colisurfaces using outer membrane protein W as an anchoring motif for the first time. Localization of catecholamines on the cell surface was confirmed by Western blot and immunofluorescence microscopy. Furthermore, cell-to-cell cohesion (i.e., cellular aggregation) induced by the displayed catecholamine and synthesis of gold nanoparticles on the cell surface support functional display of adhesive catecholamines. The engineeredE. coliexhibited significant adhesion onto various material surfaces, including silica and glass microparticles, gold, titanium, silicon, poly(ethylene terephthalate), poly(urethane), and poly(dimethylsiloxane). The uniqueness of this approach utilizing the engineered stickyE. coliis that no chemistry for cell attachment are necessary, and the ability of spontaneousE. coliattachment allows one to immobilize the cells on challenging material surfaces such as synthetic polymers. Therefore, we envision that mussel-inspired catecholamine yielded stickyE. colithat can be used as a new type of engineered microbe for various emerging fields, such as whole living cell attachment on versatile material surfaces, cell-to-cell communication systems, and many others.

scholarly journals Development of a cell surface display system in Chlamydomonas reinhardtii

2022 ◽  
Vol 61 ◽  
pp. 102570
Author(s):  
João Vitor Dutra Molino ◽  
Roberta Carpine ◽  
Karl Gademann ◽  
Stephen Mayfield ◽  
Simon Sieber
Keyword(s):  
Cell Surface ◽  
Chlamydomonas Reinhardtii ◽  
Surface Display ◽  
Cell Surface Display ◽  
Display System ◽  
A Cell ◽  
Cell Surface Display System

scholarly journals Construction of a food-grade cell surface display system for Lactobacillus casei

2014 ◽  
Vol 169 (9-10) ◽  
pp. 733-740 ◽  
Author(s):  
Jiayang Qin ◽  
Xiuwen Wang ◽  
Jian Kong ◽  
Cuiqing Ma ◽  
Ping Xu
Keyword(s):  
Cell Surface ◽  
Lactobacillus Casei ◽  
Surface Display ◽  
Cell Surface Display ◽  
Display System ◽  
Food Grade ◽  
Cell Surface Display System

Construction of a novel system for cell surface display of heterologous proteins on Pichia pastoris

2007 ◽  
Vol 29 (10) ◽  
pp. 1561-1566 ◽  
Author(s):  
Qingjie Wang ◽  
Lei Li ◽  
Min Chen ◽  
Qingsheng Qi ◽  
Peng George Wang
Keyword(s):  
Pichia Pastoris ◽  
Cell Surface ◽  
Surface Display ◽  
Cell Surface Display ◽  
Heterologous Proteins
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