A bioorthogonal chemistry approach to detect the K1 polysialic acid capsule in <i>Escherichia coli</i>

Rigolot, Vincent; Rossez, Yannick; Biot, Christophe; Lion, Cédric

doi:10.1039/d2cb00219a

Cited by 3 publications

(5 citation statements)

References 51 publications

(103 reference statements)

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“…N-Acetylmannosamine (ManNAc), precursor of K1 capsule, was armed with alkyne and azide counterpart and incorporated into bacteria and later conjugated with uorophore labelling the bacterial capsule through bio-orthogonal click chemistry and enhancing the tracing of bacteria. 158 Fukushima et al reported strain promoted double azide addition reaction producing two triazole rings in the same molecule. They made use of octadehydrodibenzo [12] annulenes (DBAs), derivative of 1,2-diethynylbenzene, and reacted it with organic azides producing bis-triazole (Fig.…”

Section: Labellingmentioning

confidence: 99%

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Clicking in harmony: exploring the bio-orthogonal overlap in click chemistry

Mehak,

Singh,

Singh

et al. 2024

RSC Adv.

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Section: Labellingmentioning

confidence: 99%

“…N -Acetylmannosamine (ManNAc), precursor of K1 capsule, was armed with alkyne and azide counterpart and incorporated into bacteria and later conjugated with fluorophore labelling the bacterial capsule through bio-orthogonal click chemistry and enhancing the tracing of bacteria. 158…”

Section: Applications Of Bio-orthogonal Click Chemistrymentioning

confidence: 99%

Clicking in harmony: exploring the bio-orthogonal overlap in click chemistry

Mehak,

Singh,

Singh

et al. 2024

RSC Adv.

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“…Rough-type lipopolysaccharides were isolated from E. coli cell cultures following an established protocol 4 . Prewarmed LB cultures (50 mL) with ampicillin (100 mg/L), arabinose (20 mg/L), and GlcNAz (0-1 mM) were inoculated with starter cultures of E. coli containing the pBAD(nahK) plasmid: the wild-type BW25113 or the ECA-deficient strain AB113 rfe::Tn10.…”

Section: Lipopolysaccharide Isolation and Analysismentioning

confidence: 99%

“…The Metabolic Usage and Glycan Destinations of GlcNAz in E. coli Alexander Eddenden 1 , Manoj Dooda 2 , Zachary A. Morrison 1 , Adithya Shankara Subramanian 3,4 , P. Lynne Howell 3,4…”

Section: Supporting Information Formentioning

confidence: 99%

The Metabolic Usage and Glycan Destinations of GlcNAz inE. coli

Eddenden,

Dooda,

Morrison

et al. 2023

Preprint

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Bacteria use a diverse range of carbohydrates to generate a profusion of glycans, with amino sugars such as N-acetylglucosamine (GlcNAc) being prevalent in the cell wall and in many exopolysaccharides. The primary substrate for GlcNAc-containing glycans, UDP-GlcNAc, is the product of the bacterial hexosamine pathway, and a key target for bacterial metabolic glycan engineering. Using the strategy of expressing NahK, to circumvent the hexosamine pathway, it is possible to directly feed the analogue of GlcNAc, N-azidoacetylglucosamine (GlcNAz, for metabolic labelling in E. coli. The cytosolic production of UDP-GlcNAz was confirmed using fluorescence assisted polyacrylamide gel electrophoresis. The key question of how GlcNAz is incorporated, was interrogated by analyzing potential sites of incorporation including peptidoglycan (PGN), the biofilm-related exopolysaccharide poly-β-1,6-N-acetylglucosamine (PNAG), as well as lipopolysaccharide (LPS) and the enterobacterial common antigen (ECA). The highest levels of incorporation were observed in PGN with lower levels in PNAG and no observable incorporation in LPS or ECA. The promiscuity of the PNAG synthase (PgaCD) towards UDP-GlcNAz in vitro and lack of undecaprenyl-pyrophosphoryl-GlcNAz intermediates generated in vivo confirmed the incorporation preferences. The results of this work will guide future developments of carbohydrate-based probes and metabolic engineering strategies.

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“…Analogues of rare deoxy amino l -sugars such as N -acetylrhamnosamine and N -acetylquinovosamine can also be tracked to their glycan destinations to discriminate between bacterial species and/or strains . Additionally, synthetic precursors for sialic, pseudaminic, , and legionaminic acids , have been shown to incorporate into their respective glycoconjugates despite requiring several enzymatic steps to generate the nucleotide sugar donors. For a more detailed discussion on chemical reporters for bacterial glycans, the reader is directed to a recent review …”

Section: Introductionmentioning

confidence: 99%

Metabolic Usage and Glycan Destinations of GlcNAz in E. coli

Eddenden,

Dooda,

Morrison

et al. 2023

ACS Chem. Biol.

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Bacteria use a diverse range of carbohydrates to generate a profusion of glycans, with amino sugars, such as Nacetylglucosamine (GlcNAc), being prevalent in the cell wall and in many exopolysaccharides. The primary substrate for GlcNAccontaining glycans, UDP-GlcNAc, is the product of the bacterial hexosamine pathway and a key target for bacterial metabolic glycan engineering. Using the strategy of expressing NahK, to circumvent the hexosamine pathway, it is possible to directly feed the analogue of GlcNAc, N-azidoacetylglucosamine (GlcNAz), for metabolic labeling in Escherichia coli. The cytosolic production of UDP-GlcNAz was confirmed by using fluorescence-assisted polyacrylamide gel electrophoresis. The key question of where GlcNAz is incorporated was interrogated by analyzing potential sites including peptidoglycan (PGN), the biofilm-related exopolysaccharide poly-β-1,6-N-acetylglucosamine (PNAG), lipopolysaccharide (LPS), and the enterobacterial common antigen (ECA). The highest levels of incorporation were observed in PGN with lower levels in PNAG and no observable incorporation in LPS or ECA. The promiscuity of the PNAG synthase (PgaCD) toward UDP-GlcNAz in vitro and the lack of undecaprenyl-pyrophosphoryl-GlcNAz intermediates generated in vivo confirmed the incorporation preferences. The results of this work will guide the future development of carbohydrate-based probes and metabolic engineering strategies.

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A bioorthogonal chemistry approach to detect the K1 polysialic acid capsule in Escherichia coli

Abstract: Most Escherichia coli strains associated with neonatal meningitis express the K1 capsule, a sialic acid polysaccharide that is directly related to their pathogenicity. Metabolic oligosaccharide engineering (MOE) has mostly been...

Cited by 3 publications

References 51 publications

Clicking in harmony: exploring the bio-orthogonal overlap in click chemistry

Clicking in harmony: exploring the bio-orthogonal overlap in click chemistry

The Metabolic Usage and Glycan Destinations of GlcNAz inE. coli

Metabolic Usage and Glycan Destinations of GlcNAz in E. coli

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