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In Vitro Assay of Bacterial Adhesion onto Mammalian Epithelial Cells

In Vitro Assay of Bacterial Adhesion onto Mammalian Epithelial Cells

Published on: May 16, 2011

Control of bacteria adhesion by cell-wall engineering.

Reiko Sadamoto¹, Kenichi Niikura, Taichi Ueda

¹Shionogi Laboratory of Biomolecular Chemistry, Hokkaido University, Kita 21 Nishi 8, Kita-ku, Sapporo 001-0021, Japan.

Journal of the American Chemical Society

|March 25, 2004

Summary

Researchers engineered bacteria to display sugars on their surface using novel cell-wall precursors. This modification enhanced bacterial adhesion, demonstrating a new method for bacterial surface engineering.

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Biomimetic Materials to Characterize Bacteria-host Interactions

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Area of Science:

Microbiology and synthetic biology
Biotechnology and biomaterials

Background:

Bacterial cell-wall biosynthesis is crucial for bacterial structure and function.
Surface display of molecules on bacteria can be achieved through various engineering strategies.

Purpose of the Study:

To develop a method for displaying specific oligosaccharides on the surface of living bacteria.
To investigate the impact of surface-displayed sugars on bacterial adhesion properties.

Main Methods:

Synthesis of UDP-MurNAc-pentapeptide derivative precursors modified with a ketone moiety.
Incubation of Lactobacilli with ketone-modified precursors to achieve surface display via cell-wall biosynthesis.
Coupling of oligomannose to the ketone moiety on the bacterial surface using an aminooxyl linker.
Assessment of bacterial adhesion using microscopic observation and surface plasmon resonance (SPR) measurements.
Evaluation of precursor incorporation enhancement with fosfomycin, a cell-wall biosynthesis inhibitor.

Main Results:

Successful display of ketone moieties on the Lactobacilli surface through cell-wall biosynthesis.
Effective surface display of oligomannose on bacteria via conjugation to the ketone moiety.
Significantly increased adhesion of sugar-displaying bacteria to concanavalin A-coated surfaces compared to native bacteria.
Enhanced incorporation of artificial cell-wall precursors in the presence of fosfomycin.

Conclusions:

The developed method enables the display of specific oligosaccharides on living bacterial surfaces.
Surface display of sugars enhances bacterial adhesion capabilities.
This approach offers a versatile platform for bacterial surface engineering with potential applications in diagnostics and therapeutics.