Bacterial cell wall quantification by a modified low-volume Nelson-Somogyi method and its use with different sugars.

Can J Microbiol

Depto. Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México.

Published: April 2022

The study of peptidoglycan-binding proteins frequently requires in vitro binding assays, in which the isolated peptidoglycan used as a substrate must be carefully quantified. Here, we describe an easy and sensitive assay for peptidoglycan quantification based on a modified Nelson-Somogyi reducing sugar assay. We report the response of this assay to different common sugars and adapt its use to peptidoglycan samples subjected to acid hydrolysis. This method showed better sensitivity than the peptidoglycan quantification method based on the acid detection of diaminopimelic acid. The method described in this work, besides being valuable in the characterization of peptidoglycan-binding proteins, is also useful for the quantification of reducing monosaccharides or polysaccharides after acid or hydrolysis.

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http://dx.doi.org/10.1139/cjm-2021-0238DOI Listing

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Article Synopsis
  • Endolysins from bacteriophages can break down bacterial cell walls and are used in various industries to combat biofilms and infections.
  • The study focused on understanding how single-domain endolysins bind to peptidoglycan, using computational methods like molecular docking and bioinformatics, which are easier compared to experimental methods.
  • The research found that Autodock Vina and the 3D-RISM module supported prior findings on the binding mechanism of a specific endolysin, showing that both computational tools effectively predicted the binding and interaction of endolysins with peptidoglycan.
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Methicillin-resistant Staphylococcus spp. present challenges in clinical and veterinary settings because effective antimicrobial agents are limited. Phage-encoded peptidoglycan-degrading enzyme, endolysin, is expected to be a novel antimicrobial agent.

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