Bacterial Cell Surface Display

Parwin, Shabnam; Kalan, Sashi; Srivastava, Preeti

doi:10.1021/bk-2019-1329.ch005

Cited by 6 publications

(4 citation statements)

References 196 publications

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“…Genetically modified organisms for usage as biosorbent are constructed by display of proteins on the cell surface. For Gram-negative bacteria, cell surface display is most often described for fusion proteins with outer membrane proteins and porins (Omp, LamB), fimbriae, pili, nucleation proteins, surface layer proteins and the maltose-binding protein [6,77]. Different metallothioneins and polyhistidines were displayed on the porins LamB [78][79][80], OmpA [81], OmpC [82] and on surface layer proteins [83].…”

Section: Biosorption and Bioaccumulationmentioning

confidence: 99%

The Advent of Biotechnology in Resource Technology

Braun,

Lederer

2023

Preprint

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The usage of biotechnological molecules and processes is an indispensable part of the resource technology research. The development of novel biomolecules therefore needs to be adapted to the existing applicational conditions and biological processes are to be examined. Novel biosorbents may not only contribute to future applications but can be of further used, e.g. to better understand the interaction of known metal- and metalloid-interacting biological molecules and materials. This review highlights biotechnological approaches published so far for bioremediation as well as metal recovery with a special spotlight on cobalt, nickel and arsenic.

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Section: Biosorption and Bioaccumulationmentioning

confidence: 99%

The Advent of Biotechnology in Resource Technology

Braun,

Lederer

2023

Preprint

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“…Surface display is a biotechnological method that involves the molecular engineering of cellular surfaces to immobilize peptides/proteins on the plasma membrane and display them in the extracellular space. This approach has been successful in displaying different biomolecules, from peptides to enzymes and antibodies, using bacteria (e.g., Escherichia coli, referred to as bacterial display) and yeast cells (e.g., Saccharomyces cerevisiae and Pichia pastoris, referred to as yeast display) as hosts [1][2][3][4]. In particular, peptides and proteins to be displayed must be fused to an anchoring motif (carrier) to ensure surface expression.…”

Section: Introductionmentioning

confidence: 99%

“…Diverse systems have been developed to display proteins on bacterial surfaces, such as those using either outer membrane protein A (OmpA) or adhesin involved in diffusible adhesion-I (AIDA-I) from E. coli, ice nucleation protein (INP) homologs from ina + bacteria (e.g., Pseudomonas syringae), and protein A from Staphylococcus aureus as anchoring motifs [3,6,7]. Among these, the Lpp-OmpA fusion remains the most popular [35,36] and has been successfully used to display a wide range of active enzymes, including βlactamases and lipases, on the surface of E. coli [37,38].…”

Section: Introductionmentioning

confidence: 99%

Surface Engineering of Escherichia coli to Display Its Phytase (AppA) and Functional Analysis of Enzyme Activities

Muñoz-Muñoz,

Terán-Ramírez,

Mares-Alejandre

et al. 2024

CIMB

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Escherichia coli phytase (AppA) is widely used as an exogenous enzyme in monogastric animal feed mainly because of its ability to degrade phytic acid or its salt (phytate), a natural source of phosphorus. Currently, successful recombinant production of soluble AppA has been achieved by gene overexpression using both bacterial and yeast systems. However, some methods for the biomembrane immobilization of phytases (including AppA), such as surface display on yeast cells and bacterial spores, have been investigated to avoid expensive enzyme purification processes. This study explored a homologous protein production approach for displaying AppA on the cell surface of E. coli by engineering its outer membrane (OM) for extracellular expression. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of total bacterial lysates and immunofluorescence microscopy of non-permeabilized cells revealed protein expression, whereas activity assays using whole cells or OM fractions indicated functional enzyme display, as evidenced by consistent hydrolytic rates on typical substrates (i.e., p-nitrophenyl phosphate and phytic acid). Furthermore, the in vitro results obtained using a simple method to simulate the gastrointestinal tract of poultry suggest that the whole-cell biocatalyst has potential as a feed additive. Overall, our findings support the notion that biomembrane-immobilized enzymes are reliable for the hydrolysis of poorly digestible substrates relevant to animal nutrition.

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“…In general, most structural studies indicate that the extracellular loops are modifiable without compromising the stability of the ß-barrel. This feature makes the loops of OMPs an attractive target for genetic modifications, with several possible biotechnological applications ( Parwin et al, 2019 ). Loop modifications have been successfully used for surface display of epitopes ( Lång, 2000 ; Rice et al, 2006 ), for the bio-adsorption of metals ( Xu and Lee, 1999 ), and for the display of trypsin cleavage sites ( Koebnik and Braun, 1993 ; Ried et al, 1994 ), all without causing any significant perturbations to the ß-barrel structure.…”

Section: Introductionmentioning

confidence: 99%

The Role of Extracellular Loops in the Folding of Outer Membrane Protein X (OmpX) of Escherichia coli

Hermansen

Ryoo

Orwick‐Rydmark

et al. 2022

Front. Mol. Biosci.

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The outer membrane of Gram-negative bacteria acts as an additional diffusion barrier for solutes and nutrients. It is perforated by outer membrane proteins (OMPs) that function most often as diffusion pores, but sometimes also as parts of larger cellular transport complexes, structural components of the cell wall, or even as enzymes. These OMPs often have large loops that protrude into the extracellular environment, which have promise for biotechnological applications and as therapeutic targets. Thus, understanding how modifications to these loops affect OMP stability and folding is critical for their efficient application. In this work, the small outer membrane protein OmpX was used as a model system to quantify the effects of loop insertions on OMP folding and stability. The insertions were varied according to both hydrophobicity and size, and their effects were determined by assaying folding into detergent micelles in vitro by SDS-PAGE and in vivo by isolating the outer membrane of cells expressing the constructs. The different insertions were also examined in molecular dynamics simulations to resolve how they affect OmpX dynamics in its native outer membrane. The results indicate that folding of OMPs is affected by both the insert length and by its hydrophobic character. Small insertions sometimes even improved the folding efficiency of OmpX, while large hydrophilic inserts reduced it. All the constructs that were found to fold in vitro could also do so in their native environment. One construct that could not fold in vitro was transported to the OM in vivo, but remained unfolded. Our results will help to improve the design and efficiency of recombinant OMPs used for surface display.

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Bacterial Cell Surface Display

Cited by 6 publications

References 196 publications

The Advent of Biotechnology in Resource Technology

The Advent of Biotechnology in Resource Technology

Surface Engineering of Escherichia coli to Display Its Phytase (AppA) and Functional Analysis of Enzyme Activities

The Role of Extracellular Loops in the Folding of Outer Membrane Protein X (OmpX) of Escherichia coli

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