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. 2022 May 4:13:885092.
doi: 10.3389/fmicb.2022.885092. eCollection 2022.

Antibacterial and Antibiofilm Activities of Chlorogenic Acid Against Yersinia enterocolitica

Kun Chen  1 , Chuantao Peng  1   2 , Fang Chi  3 , Chundi Yu  1 , Qingli Yang  1 , Zhaojie Li  1   2
Affiliations

Antibacterial and Antibiofilm Activities of Chlorogenic Acid Against Yersinia enterocolitica

Kun Chen et al. Front Microbiol. .

Abstract

Nowadays, developing new and natural compounds with antibacterial activities from plants has become a promising approach to solve antibiotic resistance of pathogenic bacteria. Chlorogenic acid (CA), as a kind of phenolic acid existing in many plants, has been found to process multifunctional activities including antibacterial activity. Herein, the antibacterial and antibiofilm activities of CA against Yersinia enterocolitica (Y. enterocolitica) were tested for the first time, and its mechanism of action was investigated. It was demonstrated that CA could exert outstanding antibacterial activity against Y. enterocolitica. Biofilm susceptibility assays further indicated that CA could inhibit biofilm formation and decrease the established biofilm biomass of Y. enterocolitica. It was deduced that through binding to Y. enterocolitica, CA destroyed the cell membrane, increased the membrane permeability, and led to bacterial cell damage. In addition, the transcriptomic analysis revealed that CA could disorder many physiological pathways, mainly including the ones of antagonizing biofilms and increasing cell membrane permeability. Finally, the spiked assay showed that the growth of Y. enterocolitica in milk was significantly inhibited by CA. Taken together, CA, as an effective bactericidal effector with application potential, exerts antagonistic activity against Y. enterocolitica by mainly intervening biofilm formation and membrane permeability-related physiological pathways.

Keywords: Yersinia enterocolitica; antibacterial activity; antibiofilm activity; antibiotic resistance; chlorogenic acid.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Antibacterial effect of CA against Y. enterocolitica. (A) Inhibition zones by water (1), 40 μg/mL gentamicin (2), 80 mg/mL CA (3), 60 mg/mL CA (4), 40 mg/mL CA (5), and 20 mg/mL CA (6). (B) Inhibition zones by water (1), 40 μg/mL gentamicin (2), 80 mg/mL CA (3), and PBS with pH of 2.5 (4). (C) Growth curves of Y. enterocolitica treated with water, gentamicin, and different concentrations of CA.
FIGURE 2
FIGURE 2
The biofilm susceptibility of Y. enterocolitica affected by CA. (A) The effects of different concentrations of CA on established biofilm of Y. enterocolitica. (B) The effects of different concentrations of CA on biofilm formation of Y. enterocolitica.
FIGURE 3
FIGURE 3
(A,B) Scanning electron microscopy (SEM) and (C,D) Transmission electron microscopy (TEM) images of Y. enterocolitica before and after treatment with CA.
FIGURE 4
FIGURE 4
Flow cytometry plots of Y. enterocolitica after CA treatment.
FIGURE 5
FIGURE 5
LSCM images of Y. enterocolitica after CA treatment.
FIGURE 6
FIGURE 6
Verification of the binding of CA to Y. enterocolitica. (A) The frequency changes during binding. (B) The weight changes during binding. (C) The fluorescence intensities of Y. enterocolitica after treatment with different concentrations of CA.
FIGURE 7
FIGURE 7
Transcriptome analysis of Y. enterocolitica after treatment with CA. (A) Volcano plot of expression of genes in Y. enterocolitica after treatment with CA. (B) Analysis of DEGs in Y. enterocolitica after treatment with CA. (a) Two-component system. (b) Biofilm formation. (c) ABC transporters.
FIGURE 8
FIGURE 8
The growth inhibition effect of CA on Y. enterocolitica in milk.
FIGURE 9
FIGURE 9
The scheme of antibacterial mechanism of CA against Y. enterocolitica.
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