title: Cell-wall engineering of living bacteria.
abstract: 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.
authors.affiliation: Sapporo Laboratory for Glycocluster Project, Japan Bioindustry Association, N10 W8, Kita-ku, Sapporo 060-0810, Japan.
authors.forename: Reiko
authors.initials: R
authors.lastname: Sadamoto
authors.forename: Kenichi
authors.initials: K
authors.lastname: Niikura
authors.forename: Pamela S
authors.initials: PS
authors.lastname: Sears
authors.forename: Haitian
authors.initials: H
authors.lastname: Liu
authors.forename: Chi-Huey
authors.initials: CH
authors.lastname: Wong
authors.forename: Akarat
authors.initials: A
authors.lastname: Suksomcheep
authors.forename: Fusao
authors.initials: F
authors.lastname: Tomita
authors.forename: Kenji
authors.initials: K
authors.lastname: Monde
authors.forename: Shin-Ichiro
authors.initials: S
authors.lastname: Nishimura
mesh_terms: 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
chemical_list: D001426:Bacterial Proteins; D019344:Lactic Acid; D019793:Fluorescein