4.5 Article

Investigating Peptidoglycan Recycling Pathways in Tannerella forsythia with N-Acetylmuramic Acid Bioorthogonal Probes

Journal

ACS INFECTIOUS DISEASES
Volume -, Issue -, Pages -

Publisher

AMER CHEMICAL SOC
DOI: 10.1021/acsinfecdis.2c00333

Keywords

peptidoglycan; bacterial cell-wall remodeling; N-acetylmuramic acid probes; bioorthogonal; bacterial pathogenesis; oral microbiome

Funding

  1. NIH U01 Common Fund Program [U01CA221230-01]
  2. NIH/NIDCR [DE029497]
  3. Delaware COBRE program
  4. National Institute of General Medical Sciences (NIGMS) [1 P30 GM110758, 1 P20 GM104316-01A1]
  5. Pew Biomedical Scholar, Sloan Scholar, and Camille Dreyfus Scholar
  6. Pew Foundation
  7. Sloan Foundation for Science Advancement
  8. Dreyfus foundation
  9. NIH
  10. Chemistry-Biology Interface (CBI) [T32GM133395]
  11. University of Delaware University Scholars Award
  12. University of Delaware Summer Scholars program
  13. David Heitzer Award

Ask authors/readers for more resources

In this study, the recycling potential of Tannerella forsythia for NAM uptake was explored. By modifying the cell wall structure and incorporating NAM analogues, the visualization of peptidoglycan was achieved. Furthermore, homologues of NAM recycling enzymes were identified in T. forsythia and successfully transformed into Escherichia coli, providing a useful tool for studying NAM recycling and peptidoglycan scavenging pathways.
The human oral microbiome is the second largest microbial community in humans, harboring over 700 bacterial species, which aid in digestion and protect from growth of disease causing pathogens. One such oral pathogen, Tannerella forsythia, along with other species, contributes to the pathogenesis of periodontitis. T. forsythia is unable to produce its own N- acetylmuramic acid (NAM) sugar, essential for peptidoglycan biosynthesis and therefore must scavenge NAM from other species with which it cohabitates. Here, we explore the recycling potential of T. forsythia for NAM uptake with a bioorthogonal modification into its peptidoglycan, allowing for click-chemistry-based visualization of the cell wall structure. Additionally, we identified NAM recycling enzyme homologues in T. forsythia that are similar to the enzymes found in Pseudomonas putida. These homologues were then genetically transformed into a laboratory safe Escherichia coli strain, resulting in the efficient incorporation of unnatural NAM analogues into the peptidoglycan backbone and its visualization, alone or in the presence of human macrophages. This strain will be useful in further studies to probe NAM recycling and peptidoglycan scavenging pathways of T. forsythia and other cohabiting bacteria.

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