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Targeting microbial biofilms using Ficin, a nonspecific plant protease

View Article: PubMed Central - PubMed

ABSTRACT

Biofilms, the communities of surface-attached bacteria embedded into extracellular matrix, are ubiquitous microbial consortia securing the effective resistance of constituent cells to environmental impacts and host immune responses. Biofilm-embedded bacteria are generally inaccessible for antimicrobials, therefore the disruption of biofilm matrix is the potent approach to eradicate microbial biofilms. We demonstrate here the destruction of Staphylococcus aureus and Staphylococcus epidermidis biofilms with Ficin, a nonspecific plant protease. The biofilm thickness decreased two-fold after 24 hours treatment with Ficin at 10 μg/ml and six-fold at 1000 μg/ml concentration. We confirmed the successful destruction of biofilm structures and the significant decrease of non-specific bacterial adhesion to the surfaces after Ficin treatment using confocal laser scanning and atomic force microscopy. Importantly, Ficin treatment enhanced the effects of antibiotics on biofilms-embedded cells via disruption of biofilm matrices. Pre-treatment with Ficin (1000 μg/ml) considerably reduced the concentrations of ciprofloxacin and bezalkonium chloride required to suppress the viable Staphylococci by 3 orders of magnitude. We also demonstrated that Ficin is not cytotoxic towards human breast adenocarcinoma cells (MCF7) and dog adipose derived stem cells. Overall, Ficin is a potent tool for staphylococcal biofilm treatment and fabrication of novel antimicrobial therapeutics for medical and veterinary applications.

No MeSH data available.


Evaluation of matrix proteins hydrolysis with Ficin.Bacteria were grown on BM medium for 72 h to form a biofilm, then a medium was replaced by the fresh one containing Ficin (1000 μg/ml) and Congo red and incubation was continued for the next 24 h.
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f3: Evaluation of matrix proteins hydrolysis with Ficin.Bacteria were grown on BM medium for 72 h to form a biofilm, then a medium was replaced by the fresh one containing Ficin (1000 μg/ml) and Congo red and incubation was continued for the next 24 h.

Mentions: To test the hydrolysis of the protein components of the biofilm matrix by Ficin, the preformed 3 day-old biofilms were treated with enzyme in the presence of Congo red, a specific dye staining the amyloid proteins (Fig. 3). The control wells incubated with Congo red in the absence of Ficin were red-stained. In the presence of Ficin a significant decrease of the staining intensity could be observed for both S. aureus and S. epidermidis plates, indicating the degradation of the protein backbone of the biofilm.


Targeting microbial biofilms using Ficin, a nonspecific plant protease
Evaluation of matrix proteins hydrolysis with Ficin.Bacteria were grown on BM medium for 72 h to form a biofilm, then a medium was replaced by the fresh one containing Ficin (1000 μg/ml) and Congo red and incubation was continued for the next 24 h.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5384253&req=5

f3: Evaluation of matrix proteins hydrolysis with Ficin.Bacteria were grown on BM medium for 72 h to form a biofilm, then a medium was replaced by the fresh one containing Ficin (1000 μg/ml) and Congo red and incubation was continued for the next 24 h.
Mentions: To test the hydrolysis of the protein components of the biofilm matrix by Ficin, the preformed 3 day-old biofilms were treated with enzyme in the presence of Congo red, a specific dye staining the amyloid proteins (Fig. 3). The control wells incubated with Congo red in the absence of Ficin were red-stained. In the presence of Ficin a significant decrease of the staining intensity could be observed for both S. aureus and S. epidermidis plates, indicating the degradation of the protein backbone of the biofilm.

View Article: PubMed Central - PubMed

ABSTRACT

Biofilms, the communities of surface-attached bacteria embedded into extracellular matrix, are ubiquitous microbial consortia securing the effective resistance of constituent cells to environmental impacts and host immune responses. Biofilm-embedded bacteria are generally inaccessible for antimicrobials, therefore the disruption of biofilm matrix is the potent approach to eradicate microbial biofilms. We demonstrate here the destruction of Staphylococcus aureus and Staphylococcus epidermidis biofilms with Ficin, a nonspecific plant protease. The biofilm thickness decreased two-fold after 24 hours treatment with Ficin at 10 μg/ml and six-fold at 1000 μg/ml concentration. We confirmed the successful destruction of biofilm structures and the significant decrease of non-specific bacterial adhesion to the surfaces after Ficin treatment using confocal laser scanning and atomic force microscopy. Importantly, Ficin treatment enhanced the effects of antibiotics on biofilms-embedded cells via disruption of biofilm matrices. Pre-treatment with Ficin (1000 μg/ml) considerably reduced the concentrations of ciprofloxacin and bezalkonium chloride required to suppress the viable Staphylococci by 3 orders of magnitude. We also demonstrated that Ficin is not cytotoxic towards human breast adenocarcinoma cells (MCF7) and dog adipose derived stem cells. Overall, Ficin is a potent tool for staphylococcal biofilm treatment and fabrication of novel antimicrobial therapeutics for medical and veterinary applications.

No MeSH data available.