Antibacterial activity of a complex bacteriocin secreted by Staphylococcus epidermidis against Porphyromonas gingivalis

Elsevier

Available online 16 May 2023, 105730

Archives of Oral BiologyAuthor links open overlay panel, , , ABSTRACTObjective

To characterize the inhibitory activity of a novel bacteriocin produced by Staphylococcus epidermidis against this periodontal pathogen.

Design

The bacteriocin activity was evaluated by the agar diffusion method over a lawn of P. gingivalis ATCC 33277. The bacteriocin was purified by Reverse Phase-High Performance Liquid Chromatography (RP-HPLC) and Matrix Assisted Laser Desorption Ionization -Time of Flight Mass Spectrometry (MALDI-TOF-MS). In addition, the bacteriocin host specificity, production on different media cultures and susceptibility to enzymes, pH, and heat treatment were determined.

Results

The bacteriocin BAC 14990 was selective to P. gingivalis, suggesting a narrow activity range. The production during the growth curve indicated that S. epidermidis had a continued production of this antimicrobial, showing the highest concentration in the stationary phase. The purification of BAC 14990 showed that bacteriocin had a molecular mass of 5795 Da. BAC 14990 was partially resistant to the treatment with proteinase K and papain, however, was fully susceptible to amylase treatment indicating the presence of sugar residues in the protein, suggesting a conjugated type of bacteriocin. Also, this diffusible inhibitory substance was heat and pH treatment resistant.

Conclusions

The results indicate the isolation of a new staphylococcal complex bacteriocin that is able to eliminate a Gram-negative bacterium. These results could contribute to the development of treatments directed against pathogens in mixed communities, as is the case with oral diseases.

Section snippetsINTRODUCTION

The oral cavity consists of diverse microbiome with more than 700 bacterial species (Gao et al., 2018). These bacteria play an important role both in maintaining oral health and in the etiology of oral diseases (Kilian et al., 2016). Periodontitis is one of the most prevalent diseases in the world population and is produced by certain microorganisms contained in dental plaque (Nazir et al., 2020), that under certain conditions increase in number and proportion. Porphyromonas gingivalis is an

Bacterial strains and growth media

The bacterial strains used for the screening of bacteriocins production and evaluation for antimicrobial activities are shown in Table 1. Staphylococcus spp., Streptococcus spp. and bacteria belonging to the Enterobacteriaceae family were grown in Tryptic Soy broth (Merck) at 37ºC for 18-24 h. Aggregatibacter actinomycetemcomytans was grown in Brain Heart Infusion (Becton and Dickinson) at 37ºC in a humidified, 5% CO2 atmosphere. Porphyromonas gingivalis, Porphyromonas endodontalis, Prevotella

Bacteriocin-producing bacteria

We analyzed the growth inhibitory capacity of different reference strains (ATCC) and clinical isolates on P. gingivalis. Staphylococcus epidermidis ATCC 14990 and 6 clinical isolates had an inhibitory effect on P. gingivalis (Fig.1 and, Table 1). These clinical isolates were identified as S. epidermidis, Lactobacillus paracasei and Prevotella nigrescens. Previous reports have identified bacteriocin against P. gingivalis produced by P. nigrescens, L. paracasei and L. plantarum NC8 (Khalaf et

DISCUSSION

P. gingivalis is one of the etiologic agents responsible of periodontitis. Dental microbiome is composed for more 700 species forming dental plaque which is a polymicrobial community embedded in a complex extracellular matrix (Gao et al., 2018). This complex community principally contains Firmicutes, Bacteroidetes, Proteobacteria, Actinobacteria, Spirochaetes and Fusobacteria (Alghamdi, 2022). Within the biofilm there are beneficial relationships between the microorganisms, but also have

CRediT authorship contribution statement

Mario Castillo-Ruiz: Investigation, Conceptualization, Methodology, Writing-Original draft preparation. Leslie Daile: Visualization, Writing-Reviewing and Editing. Pamela Machuca: Investigation, Writing-Reviewing and Editing. Mauricio Bittner: Conceptualization, Supervision, Writing-Reviewing and Editing

Declaration of Competing Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

ACKNOWLEDGEMENTS

This study was supported by UNAB grant 48-09/R, UNAB grant DI 24-09/I. The founders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The authors declare that there are no conflicts of interest in this study. We thank Alejandro González-Candia for technical support with the experiments.

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