Utilization of different MurNAc sources by the oral pathogen Tannerella forsythia and role of the inner membrane transporter AmpG.

Autor: Mayer VMT; Department of NanoBiotechnology, NanoGlycobiology unit, Universität für Bodenkultur Wien, Vienna, Austria., Tomek MB; Department of NanoBiotechnology, NanoGlycobiology unit, Universität für Bodenkultur Wien, Vienna, Austria., Figl R; Department of Chemistry, Institute of Biochemistry, Universität für Bodenkultur Wien, Vienna, Austria., Borisova M; Department of Biology, Eberhard Karls Universität Tübingen, Microbiology/Glycobiology, Interfaculty Institute of Microbiology and Infection Medicine Tübingen, Tübingen, Germany., Hottmann I; Department of Biology, Eberhard Karls Universität Tübingen, Microbiology/Glycobiology, Interfaculty Institute of Microbiology and Infection Medicine Tübingen, Tübingen, Germany., Blaukopf M; Department of Chemistry, Institute of Organic Chemistry, Universität für Bodenkultur Wien, Vienna, Austria., Altmann F; Department of Chemistry, Institute of Biochemistry, Universität für Bodenkultur Wien, Vienna, Austria., Mayer C; Department of Biology, Eberhard Karls Universität Tübingen, Microbiology/Glycobiology, Interfaculty Institute of Microbiology and Infection Medicine Tübingen, Tübingen, Germany. christoph.mayer@uni-tuebingen.de., Schäffer C; Department of NanoBiotechnology, NanoGlycobiology unit, Universität für Bodenkultur Wien, Vienna, Austria. christina.schaeffer@boku.ac.at.
Jazyk: angličtina
Zdroj: BMC microbiology [BMC Microbiol] 2020 Nov 17; Vol. 20 (1), pp. 352. Date of Electronic Publication: 2020 Nov 17.
DOI: 10.1186/s12866-020-02006-z
Abstrakt: Background: The Gram-negative oral pathogen Tannerella forsythia strictly depends on the external supply of the essential bacterial cell wall sugar N-acetylmuramic acid (MurNAc) for survival because of the lack of the common MurNAc biosynthesis enzymes MurA/MurB. The bacterium thrives in a polymicrobial biofilm consortium and, thus, it is plausible that it procures MurNAc from MurNAc-containing peptidoglycan (PGN) fragments (muropeptides) released from cohabiting bacteria during natural PGN turnover or cell death. There is indirect evidence that in T. forsythia, an AmpG-like permease (Tanf_08365) is involved in cytoplasmic muropeptide uptake. In E. coli, AmpG is specific for the import of N-acetylglucosamine (GlcNAc)-anhydroMurNAc(-peptides) which are common PGN turnover products, with the disaccharide portion as a minimal requirement. Currently, it is unclear which natural, complex MurNAc sources T. forsythia can utilize and which role AmpG plays therein.
Results: We performed a screen of various putative MurNAc sources for T. forsythia mimicking the situation in the natural habitat and compared bacterial growth and cell morphology of the wild-type and a mutant lacking AmpG (T. forsythia ΔampG). We showed that supernatants of the oral biofilm bacteria Porphyromonas gingivalis and Fusobacterium nucleatum, and of E. coli ΔampG, as well as isolated PGN and defined PGN fragments obtained after enzymatic digestion, namely GlcNAc-anhydroMurNAc(-peptides) and GlcNAc-MurNAc(-peptides), could sustain growth of T. forsythia wild-type, while T. forsythia ΔampG suffered from growth inhibition. In supernatants of T. forsythia ΔampG, the presence of GlcNAc-anhMurNAc and, unexpectedly, also GlcNAc-MurNAc was revealed by tandem mass spectrometry analysis, indicating that both disaccharides are substrates of AmpG. The importance of AmpG in the utilization of PGN fragments as MurNAc source was substantiated by a significant ampG upregulation in T. forsythia cells cultivated with PGN, as determined by quantitative real-time PCR. Further, our results indicate that PGN-degrading amidase, lytic transglycosylase and muramidase activities in a T. forsythia cell extract are involved in PGN scavenging.
Conclusion: T. forsythia metabolizes intact PGN as well as muropeptides released from various bacteria and the bacterium's inner membrane transporter AmpG is essential for growth on these MurNAc sources, and, contrary to the situation in E. coli, imports both, GlcNAc-anhMurNAc and GlcNAc-MurNAc fragments.
Databáze: MEDLINE
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