Control of Bacteria Adhesion by Cell-Wall Engineering

Abstract

UDP-MurNAc-pentapeptide derivative bacterial cell-wall precursors were synthesized as effective tools for surface display on living bacteria. Lactobacilli were incubated in the ketone-modified precursor-containing medium, and the ketone moiety was displayed on the bacterial surface through cell-wall biosynthesis. Oligomannose was coupled with the ketone moiety on the bacterial surface via a aminooxyl linker, thereby displaying this oligosaccharide on the surface of the bacteria. The increase in the adhesion of the sugar-displaying bacteria onto a concanavalin A-attached film compared to that of native bacteria was confirmed by microscopic observation and surface plasmon resonance measurement. The incorporation of the artificial cell-wall precursors was enhanced when incubated with fosfomycin, an inhibitor of cell-wall precursor biosynthesis.

A Pictet-Spengler ligation for protein chemical modification

“…We focused here on the use of the Pictet-Spengler ligation for modification of purified proteins, but applications extend to other biomolecules that are amenable to functionalization with reactive carbonyl groups. Methods for metabolic (49)(50)(51), enzymatic (52), and chemical (53,54) functionalization of glycans with ketone and aldehyde groups are well-established and are finding use in proteomic analyses of glycosylated proteins. The Pictet-Spengler ligation may enhance the performance of these methods as well as others that seek to detect or manipulate carbonyl groups using bioorthogonal chemistry (55,56).…”

Click Chemistry in Complex Mixtures: Bioorthogonal Bioconjugation

“…Although they are not completely abiotic and are present in some intracellular metabolites, their synthetic accessibility and small size has made them amenable to incorporation into biomolecules. Popular routes include uptake and processing of unnatural intermediates of glycan biosynthesis (Jacobs et al, 2000; Luchansky et al, 2004; Mahal et al, 1997; Sadamoto et al, 2004), genetically-controlled incorporation into proteins in a residue-specific (Datta et al, 2002; Ngo and Tirrell, 2011; Tang et al, 2009) or site-specific manner (Carrico et al, 2007; Chin et al, 2003; Hudak et al, 2012; Hudak et al, 2011; Liu and Schultz, 2010; Wang et al, 2003a; Wu et al, 2009), and use in peptide tags that direct enzymatic ligation of aldehyde or ketone bearing small molecules (Chen et al, 2005; Rashidian et al, 2012). Aldehydes can also be introduced into proteins by periodate oxidation of N -terminal Ser/Thr residues (Geoghegan and Stroh, 1992), site-specific protein transaminations (Geoghegan et al, 1993; Gilmore et al, 2006; Witus et al, 2010; Witus et al, 2013), periodate oxidation of sialic acids displayed on proteins (Hage et al, 1997), and by addition of ketone containing small molecules to protein C-terminal thioesters generated by expressed protein ligation (Esser-Kahn and Francis, 2008).…”

Versatile microbial surface-display for environmental remediation and biofuels production

“…Another cell wall precursor UDP-MurNAc pentapeptide coupled with oligomannose was applied successfully to cell surface display [61]. Lactobacillus plantarum was grown in medium containing the bacterial cell-wall precursor and surface display of mannose was confirmed through adhesion to the ConA (mannose-bindingprotein)-immobilized surface.…”