AI Article Synopsis

  • Bacteria utilize various carbohydrates to create complex structures called glycans, particularly incorporating amino sugars like -acetylglucosamine (GlcNAc) in their cell walls and exopolysaccharides.
  • A method was developed to bypass the bacterial hexosamine pathway by expressing NahK, allowing the introduction of a GlcNAc analogue, -azidoacetylglucosamine (GlcNAz), which was confirmed to produce another compound, UDP-GlcNAz, using gel electrophoresis.
  • The study found that GlcNAz was primarily incorporated into peptidoglycan (PGN), while lower levels were seen in poly-β-1,6--acetylglucos

Article Abstract

Bacteria use a diverse range of carbohydrates to generate a profusion of glycans, with amino sugars such as -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, -azidoacetylglucosamine (GlcNAz), for metabolic labelling in The cytosolic production of UDP-GlcNAz was confirmed 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--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) towards UDP-GlcNAz and lack of undecaprenyl-pyrophosphoryl-GlcNAz intermediates generated 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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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10462111PMC
http://dx.doi.org/10.1101/2023.08.17.553294DOI Listing

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