Horseradish peroxidase interacts with the cell wall peptidoglycans on oral bacteria

Mizuno,Hirofumi; Takayama,Eiji; Satoh,Ayano; Into,Takeshi; Adachi,Masanori; Ekuni,Daisuke; Yashiro,Koji; Mizuno‑Kamiya,Masako; Nagayama,Motohiko; Saku,Seitaro; Tomofuji,Takaaki; Doi,Yutaka; Murakami,Yukitaka; Kondoh,Nobuo; Morita,Manabu

doi:10.3892/etm.2020.9016

Horseradish peroxidase interacts with the cell wall peptidoglycans on oral bacteria

Authors:
- Hirofumi Mizuno
- Eiji Takayama
- Ayano Satoh
- Takeshi Into
- Masanori Adachi
- Daisuke Ekuni
- Koji Yashiro
- Masako Mizuno‑Kamiya
- Motohiko Nagayama
- Seitaro Saku
- Takaaki Tomofuji
- Yutaka Doi
- Yukitaka Murakami
- Nobuo Kondoh
- Manabu Morita
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Affiliations: Department of Preventive Dentistry, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama 700‑8558, Japan, Department of Oral Biochemistry, Asahi University School of Dentistry, Mizuho, Gifu 501‑0296, Japan, Department of Medical Bioengineering, Graduate School of Interdisciplinary Science and Engineering in Healthcare Systems, Okayama University, Okayama 700‑8530, Japan, Department of Oral Microbiology, Asahi University School of Dentistry, Mizuho, Gifu 501‑0296, Japan, Department of Dental Materials Science, Asahi University School of Dentistry, Mizuho, Gifu 501‑0296, Japan, Department of Oral Pathology, Asahi University School of Dentistry, Mizuho, Gifu 501‑0296, Japan, Department of Operative Dentistry, Asahi University School of Dentistry, Mizuho, Gifu 501‑0296, Japan
Published online on: July 16, 2020 https://doi.org/10.3892/etm.2020.9016
Pages: 2822-2827

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Abstract

Salivary peroxidase and myeloperoxidase are known to display antibacterial activity against oral microbes, and previous indications have pointed to the possibility that horseradish peroxidase (HRP) adsorbs onto the membrane of the major oral streptococci, Streptococcus mutans and Streptococcus sanguinis (S. sanguinis). However, the mechanism of interaction between HRP and the bacterial cell wall component is unclear. Dental plaques containing salivary glycoproteins and extracellular microbial products are visualized with ‘dental plaque disclosing agent’, and are controlled within dental therapy. However, current ‘dental plaque disclosing agents’ are difficult to evaluate with just dental plaques, since they stain and disclose not only dental plaques but also pellicle formed with salivary glycoproteins on a tooth surface. In this present study, we have demonstrated that HRP interacted with the cell wall component of the major gram‑positive bacterial peptidoglycan, but not the major cell wall component of gram‑negative bacteria lipopolysaccharide. Furthermore, we observed that the adsorbed HRP labeled with fluorescence was detected on the major oral gram‑positive strains S. sanguinis and Streptococcus salivarius (S. salivarius), but not on a gram‑negative strain, Escherichia coli (E. coli). Furthermore, we have demonstrated that the combination of HRP and chromogenic substrate clearly disclosed the dental plaques and the biofilm developed by S. sanguinis, S. salivarius and the major gram‑postive bacteria Lactobacillus casei on tooth surfaces, and slightly disclosed the biofilm by E. coli. The combination of HRP and chromogenic substrate did not stain either the dental pellicle with the salivary glycoprotein mucin, or naked tooth surfaces. These results have suggested the possibility that the adsorption activity of HRP not only contributes to the evaluation of dental plaque, but that enzymatic activity of HRP may also contribute to improve dental hygiene.

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