BrkAutoDisplay: functional display of multiple exogenous proteins on the surface of Escherichia coli by using BrkA autotransporter

BackgroundBacterial surface display technique enables the exogenous proteins or polypeptides displayed on the bacterial surface, while maintaining their relatively independent spatial structures and biological activities. The technique makes recombinant bacteria possess the expectant functions, subsequently, directly used for many applications. Many proteins could be used to achieve bacterial surface display, among them, autotransporter, a member of the type V secretion system of gram-negative bacteria, has been extensively studied because of its modular structure and apparent simplicity. However, autotransporter has not been widely used at present due to lack of a convenient genetic vector system. With our recently characterized autotransporter BrkA (Bordetella serum-resistance killing protein A) from Bordetella pertussis, we are aiming to develop a new autotransporter-based surface display system for potential wide application.ResultsHere, we construct a bacterial surface display system named as BrkAutoDisplay, based on the structure of autotransporter BrkA. BrkAutoDisplay is a convenient system to host exogenous genes. In our test, this system is good to efficiently display various proteins on the outer membrane surface of Escherichia coli, including green fluorescent protein (GFP), various enzymes and single chain antibody. Moreover, the displayed GFP possesses green fluorescence, the enzymes CotA, EstPc and PalA exhibit catalytic activity 0.12, 6.88 and 0.32 mU (per 5.2 × 108 living bacteria cells) respectively, and the single chain antibody fragment (scFv) can bind with its antigen strongly. Finally, we showed that C41(DE3) is a good strain of E. coli for the successful functionality of BrkAutoDisplay.ConclusionsWe designed a new bacterial display system called as BrkAutoDisplay and displayed various exogenous proteins on E. coli surface. Our results indicate that BrkAutoDisplay system is worthy of further study for industrial applications.

• Publication year

  2015

• Venue

  Microbial Cell Factories

• Publication date

  2015-09-04

• Fields of study

  Biology, Medicine, Engineering

• Identifiers
  DOI 10.1186/s12934-015-0316-3PMID 26337099PMCID 4558763
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Autotransporter-based cell surface display in Gram-negative bacteria

  2015influential reference
• Phage display-based generation of novel internalizing antibody fragments for immunotoxin-based treatment of acute myeloid leukemia

  2015cited by this paper
• A mortise–tenon joint in the transmembrane domain modulates autotransporter assembly into bacterial outer membranes

  2014cited by this paper
• Reconstitution of bacterial autotransporter assembly using purified components

  2014cited by this paper
• Antigen 43/Fcε3 chimeric protein expressed by a novel bacterial surface expression system as an effective asthma vaccine

  2014cited by this paper
• Cell surface display of cold-active esterase EstPc with the use of a new autotransporter from Psychrobacter cryohalolentis K5T

  2014influential reference
• Autodisplay for the co-expression of lipase and foldase on the surface of E. coli: washing with designer bugs

  2014cited by this paper
• Comparison of Autotransporter and Ice Nucleation Protein as Carrier Proteins for Antibody Display on The Cell Surface of Escherichia coli

  2013cited by this paper
• Dynamic association of BAM complex modules includes surface exposure of the lipoprotein BamC.

  2012cited by this paper
• Probing the applicability of autotransporter based surface display with the EstA autotransporter of Pseudomonas stutzeri A15

  2012cited by this paper
• Autodisplay of functional CYP106A2 in Escherichia coli.

  2012cited by this paper
• From self sufficiency to dependence: mechanisms and factors important for autotransporter biogenesis

  2012cited by this paper
• Autodisplay of enzymes--molecular basis and perspectives.

  2012influential reference
• Structures and functions of autotransporter proteins in microbial pathogens.

  2011cited by this paper
• Bacterial display in combinatorial protein engineering

  2011cited by this paper
• Autotransporter passenger domain secretion requires a hydrophobic cavity at the extracellular entrance of the β-domain pore.

  2011cited by this paper
• Vectorial transport and folding of an autotransporter virulence protein during outer membrane secretion

  2009influential reference
• Differential Bacterial Surface Display of Peptides by the Transmembrane Domain of OmpA

  2009cited by this paper
• Isolation of engineered, full-length antibodies from libraries expressed in Escherichia coli

  2007cited by this paper
• Cleavage of a bacterial autotransporter by an evolutionarily convergent autocatalytic mechanism

  2007cited by this paper
• Protein secretion in gram-negative bacteria via the autotransporter pathway.

  2007cited by this paper
• Protein engineering with bacterial display.

  2007cited by this paper
• Incorporation of a polypeptide segment into the β‐domain pore during the assembly of a bacterial autotransporter

  2007cited by this paper
• Polar Localization of the Autotransporter Family of Large Bacterial Virulence Proteins

  2006cited by this paper
• Autodisplay: efficient bacterial surface display of recombinant proteins

  2006cited by this paper
• Glycosylation of the Self-Recognizing Escherichia coli Ag43 Autotransporter Protein

  2006cited by this paper
• Efficient secretion of a folded protein domain by a monomeric bacterial autotransporter

  2005cited by this paper
• Novel prenyltransferase enzymes as a tool for flavonoid prenylation.

  2005cited by this paper
• Anchored periplasmic expression, a versatile technology for the isolation of high-affinity antibodies from Escherichia coli-expressed libraries.

  2004cited by this paper
• Enoyl-CoA hydratase. reaction, mechanism, and inhibition.

  2003cited by this paper
• New technologies in therapeutic antibody development.

  2003cited by this paper
• Display and release of the Plasmodium falciparum circumsporozoite protein using the autotransporter MisL of Salmonella enterica.

  2003cited by this paper
• The pore size of the autotransporter domain is critical for the active translocation of the passenger domain.

  2003cited by this paper
• Antigen 43-Mediated Autotransporter Display, a Versatile Bacterial Cell Surface Presentation System

  2002cited by this paper
• Antibodies to BrkA augment killing of Bordetella pertussis.

  2001cited by this paper
• Recombinant antibodies for cancer diagnosis and therapy

  2001cited by this paper
• CotA of Bacillus subtilis Is a Copper-Dependent Laccase

  2001cited by this paper
• Expression of Single Chain Antibodies (ScFvs) for c‐myc Oncoprotein in Recombinant Escherichiacoli Membranes by Using the Ice‐Nucleation Protein of Pseudomonassyringae

  2000cited by this paper
• The great escape: structure and function of the autotransporter proteins.

  1998cited by this paper
• Cloning and sequencing of a Bordetella pertussis serum resistance locus

  1994cited by this paper
• AIDA‐I, the adhesin involved in diffuse adherence of the diarrhoeagenic Escherichia coli strain 2787 (O126:H27), is synthesized via a precursor molecule

  1992cited by this paper
• Gene structure and extracellular secretion of Neisseria gonorrhoeae IgA protease

  1987cited by this paper
• L-3-hydroxyacyl coenzyme A dehydrogenase from pig heart muscle. EC 1.1.1.35 L-3-hydroxyacyl-CoA: NAD oxidoreductase.

  1975cited by this paper
• [16]l-3-hydroxyacyl coenzyme A dehydrogenase from pig heart muscle

  1975cited by this paper

• The Midas touch: Recombinant antibodies transforming diagnostic platforms.

  2026cites this paper
• Lactococcus lactis as a New Strategy for Oral Vaccination: Current Insights and Future Perspectives

  2026cites this paper
• Escherichia coli Surface Display: Advances and Applications in Biocatalysis.

  2025cites this paper
• Increased Synthesis of a Magnesium Transporter MgtA During Recombinant Autotransporter Expression in Escherichia coli

  2021influential citation
• Oxygen vector accelerates farnesylation and redox reaction to promote the biosynthesis of 4-acetylantroquinonol B and antroquinonol during submerged fermentation of Antrodia cinnamomea

  2020cites this paper
• Biosynthesis of (deoxy)guanosine-5'-triphosphate by GMP kinase and acetate kinase fixed on the surface of E. coli.

  2019cites this paper
• Molecular optimization of autotransporter-based tyrosinase surface display.

  2019cites this paper
• Comparing autotransporter β-domain configurations for their capacity to secrete heterologous proteins to the cell surface

  2018cites this paper
• High-vacuum optical platform for cryo-CLEM (HOPE): A new solution for non-integrated multiscale correlative light and electron microscopy.

  2018cites this paper
• Bacillus Cellulase Molecular Cloning, Expression, and Surface Display on the Outer Membrane of Escherichia coli

  2018cites this paper
• Evolution and Engineering in Escherichia coli

  2016cites this paper
• A nanobody:GFP bacterial platform that enables functional enzyme display and easy quantification of display capacity

  2016cites this paper