Cell-wall engineering of living bacteria.
The cell walls of living bacteria were chemically modified by adding cell-wall precursors. As the precursors to be incorporated into the cell wall, UDP-MurNAc pentapeptide, lipid I, and lipid II derivatives were synthesized. The aimed compounds were attached to the amine residue of lysine at the pentapeptide moiety. Fluorescein-attached UDP-MurNAc pentapeptide was efficiently incorporated into both Gram-positive and Gram-negative bacteria. In the case of Gram-negative bacteria, such as Escherichia coli, the permeability of the outer membrane (lipopolysaccharide layer) was enhanced by EDTA treatment before the incorporation. For Gram-positive bacteria, UDP-MurNAc derivatives were incorporated in the cell wall without EDTA treatment due to the lack of the lipopolysaccharide layer. Furthermore, instead of dyes, a ketone group was attached to the UDP-MurNAc pentapeptide. The ketone group was also delivered to the bacterial cell wall of lactic acid bacteria, giving a platform to attach large molecules on the surface.
Sadamoto
Reiko
R
Sapporo Laboratory for Glycocluster Project, Japan Bioindustry Association, N10 W8, Kita-ku, Sapporo 060-0810, Japan.
Niikura
Kenichi
K
Sears
Pamela S
PS
Liu
Haitian
H
Wong
Chi-Huey
CH
Suksomcheep
Akarat
A
Tomita
Fusao
F
Monde
Kenji
K
Nishimura
Shin-Ichiro
S
D001419:Bacteria / Q000737:chemistry* / Q000648:ultrastructure; D001426:Bacterial Proteins / Q000737:chemistry*; D002473:Cell Wall / Q000737:chemistry*; D004926:Escherichia coli / Q000737:chemistry; D019793:Fluorescein; D006094:Gram-Positive Bacteria / Q000737:chemistry; D019344:Lactic Acid / Q000737:chemistry; D008856:Microscopy, Fluorescence; D013050:Spectrometry, Fluorescence
D001426:Bacterial Proteins; D019344:Lactic Acid; D019793:Fluorescein