{"title":"Antimicrobial and antibiofilm effects of a leucine-substituted rice peptide against Porphyromonas gingivalis and Fusobacterium nucleatum","authors":"Yukari Aoki-Nonaka , Aoi Matsugishi-Nasu , Hnin Yu Lwin, Yukako Minato, Daiki Ando, Yuko Warita, Naoki Takahashi, Koichi Tabeta","doi":"10.1016/j.micpath.2025.108052","DOIUrl":null,"url":null,"abstract":"<div><div>The aim of this study to elucidate the antibacterial and antibiofilm effects of a leucine-substituted peptide against periodontal pathogen. N3L peptide was designed as a leucine-substituted variant of the rice-derived AmyI-1-18 peptide. Subsequently, the minimum inhibitory concentration and minimum bactericidal concentration of N3L against <em>Porphyromonas gingivalis</em> and <em>Fusobacterium nucleatum</em> were determined. Furthermore, its membrane permeability was investigated by propidium iodide (PI) staining. Moreover, the effects of N3L on the gene expression of virulence factors were analyzed by reverse transcriptase quantitative polymerase chain reaction. The biofilm amount was assessed by crystal violet staining to analyze the biofilm control ability of N3L peptide. N3L demonstrated bactericidal activity against <em>P. gingivalis</em> and <em>F. nucleatum</em>, with notably enhanced antimicrobial activity against <em>P. gingivalis</em> compared with the parent peptide AmyI-1-18. PI staining indicated that N3L showed bacterial membrane permeability against <em>P. gingivalis</em> and <em>F. nucleatum</em>. Moreover, N3L significantly downregulated the expression levels of <em>fimA</em>, <em>rgpA</em>, <em>rgpB</em>, and <em>kgp</em> in <em>P. gingivalis</em> and <em>fadA</em> in <em>F. nucleatum</em>. This peptide also significantly inhibited biofilm formation and reduced established biofilms of <em>P. gingivalis</em> and <em>F. nucleatum</em>. The leucine-substituted rice peptide N3L, with its hydrophobicity and α-helix content higher than those of AmyI-1-18, showed enhanced antimicrobial and antibiofilm activities through membrane disruption and modulation of gene expressions related to adherence and metabolism. The leucine-substituted rice peptide N3L showed enhanced antimicrobial and antibiofilm activities against <em>P. gingivalis</em> and <em>F. nucleatum</em>.</div></div>","PeriodicalId":18599,"journal":{"name":"Microbial pathogenesis","volume":"209 ","pages":"Article 108052"},"PeriodicalIF":3.5000,"publicationDate":"2025-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Microbial pathogenesis","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0882401025007776","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"IMMUNOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
The aim of this study to elucidate the antibacterial and antibiofilm effects of a leucine-substituted peptide against periodontal pathogen. N3L peptide was designed as a leucine-substituted variant of the rice-derived AmyI-1-18 peptide. Subsequently, the minimum inhibitory concentration and minimum bactericidal concentration of N3L against Porphyromonas gingivalis and Fusobacterium nucleatum were determined. Furthermore, its membrane permeability was investigated by propidium iodide (PI) staining. Moreover, the effects of N3L on the gene expression of virulence factors were analyzed by reverse transcriptase quantitative polymerase chain reaction. The biofilm amount was assessed by crystal violet staining to analyze the biofilm control ability of N3L peptide. N3L demonstrated bactericidal activity against P. gingivalis and F. nucleatum, with notably enhanced antimicrobial activity against P. gingivalis compared with the parent peptide AmyI-1-18. PI staining indicated that N3L showed bacterial membrane permeability against P. gingivalis and F. nucleatum. Moreover, N3L significantly downregulated the expression levels of fimA, rgpA, rgpB, and kgp in P. gingivalis and fadA in F. nucleatum. This peptide also significantly inhibited biofilm formation and reduced established biofilms of P. gingivalis and F. nucleatum. The leucine-substituted rice peptide N3L, with its hydrophobicity and α-helix content higher than those of AmyI-1-18, showed enhanced antimicrobial and antibiofilm activities through membrane disruption and modulation of gene expressions related to adherence and metabolism. The leucine-substituted rice peptide N3L showed enhanced antimicrobial and antibiofilm activities against P. gingivalis and F. nucleatum.
期刊介绍:
Microbial Pathogenesis publishes original contributions and reviews about the molecular and cellular mechanisms of infectious diseases. It covers microbiology, host-pathogen interaction and immunology related to infectious agents, including bacteria, fungi, viruses and protozoa. It also accepts papers in the field of clinical microbiology, with the exception of case reports.
Research Areas Include:
-Pathogenesis
-Virulence factors
-Host susceptibility or resistance
-Immune mechanisms
-Identification, cloning and sequencing of relevant genes
-Genetic studies
-Viruses, prokaryotic organisms and protozoa
-Microbiota
-Systems biology related to infectious diseases
-Targets for vaccine design (pre-clinical studies)