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MRJP1-containing glycoproteins isolated from honey, a novel antibacterial drug candidate with broad spectrum activity against multi-drug resistant clinical isolates.

Brudzynski K, Sjaarda C, Lannigan R - Front Microbiol (2015)

Bottom Line: Their resistance to different classes of antibiotics was confirmed using automated system Vitek 2.Glps isolated from different honeys showed a similar ability to overcome bacterial resistance to β-lactams suggesting that (a) their mode of action is distinct from other classes of β-lactams and that (b) the common glps structure was the lead structure responsible for the activity.The results of the current study together with our previous evidence of a rapid bactericidal activity of glps demonstrate that glps possess suitable characteristics to be considered a novel antibacterial drug candidate.

View Article: PubMed Central - PubMed

Affiliation: Department of Drug Discovery and Development Department St. Catharines, ON, Canada.

ABSTRACT
The emergence of extended- spectrum β-lactamase (ESBL) is the underlying cause of growing antibiotic resistance among Gram-negative bacteria to β-lactam antibiotics. We recently reported the discovery of honey glycoproteins (glps) that exhibited a rapid, concentration-dependent antibacterial activity against both Gram-positive Bacillus subtilis and Gram-negative Escherichia coli that resembled action of cell wall-active β-lactam drugs. Glps showed sequence identity with the Major Royal Jelly Protein 1 (MRJP1) precursor that harbors three antimicrobial peptides: Jelleins 1, 2, and 4. Here, we used semi-quantitative radial diffusion assay and broth microdilution assay to evaluate susceptibility of a number of multi-drug resistant (MDR) clinical isolates to the MRJP1-contaning honey glycoproteins. The MDR bacterial strains comprised three methicillin-resistant Staphylococcus aureus (MRSA), four Pseudomonas aeruginosa, two Klebsiella pneumoniae, two vancomycin-resistant Enterococci (VRE), and five ESBL identified as one Proteus mirabilis, three E. coli, and one E. coli NDM. Their resistance to different classes of antibiotics was confirmed using automated system Vitek 2. MDR isolates differed in their susceptibility to glps with MIC90 values ranging from 4.8 μg/ml against B. subtilis to 14.4 μg/ml against ESBL K. pneumoniae, Klebsiella spp. ESBL and E. coli and up to 33 μg/ml against highly resistant strains of P. aeruginosa. Glps isolated from different honeys showed a similar ability to overcome bacterial resistance to β-lactams suggesting that (a) their mode of action is distinct from other classes of β-lactams and that (b) the common glps structure was the lead structure responsible for the activity. The results of the current study together with our previous evidence of a rapid bactericidal activity of glps demonstrate that glps possess suitable characteristics to be considered a novel antibacterial drug candidate.

No MeSH data available.


Related in: MedlinePlus

Linear relationship between ampicillin and glycoprotein concentrations and the diameter of the zone of inhibition. (A) Amp against E. coli, (B) Amp against B. subtilis, (C) relationship between the diameter of inhibition zone and ampicillin concentration and (D). glp G208 against E. coli, (E) glp G208 against B. subtilis, and (F) relationship between the diameter of inhibition zone and glp concentrations. Columns represent means and standard errors.
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Figure 2: Linear relationship between ampicillin and glycoprotein concentrations and the diameter of the zone of inhibition. (A) Amp against E. coli, (B) Amp against B. subtilis, (C) relationship between the diameter of inhibition zone and ampicillin concentration and (D). glp G208 against E. coli, (E) glp G208 against B. subtilis, and (F) relationship between the diameter of inhibition zone and glp concentrations. Columns represent means and standard errors.

Mentions: In vitro evaluation of susceptibility of clinical isolates was performed using a semi-quantitative radial well-diffusion and broth microdilution assay. To compare the results of bacterial susceptibility to honey glycoproteins in a quantitative manner, the reference method has been designed that established the relationship between ampicillin dilutions and zone of inhibition (ZOI) in well-diffusion assay. As shown in Figure 2, the series of twofold diluted honey glycoproteins and ampicillin (1 mg/ml stock solutions) produced concentration-dependent ZOIs with R2 = 0.99, for ampicillin and R2 = 0.96 and 0.95 for glps tested against E. coli and B. subtilis, respectively. MIC values from well-diffusion assay against E. coli and B. subtilis (1 × 106 CFU/ml) were <0.025 μg/well or <2.0 μg/ml for ampicillin and <0.046 μg/well or 5.7 μg/ml for glps.


MRJP1-containing glycoproteins isolated from honey, a novel antibacterial drug candidate with broad spectrum activity against multi-drug resistant clinical isolates.

Brudzynski K, Sjaarda C, Lannigan R - Front Microbiol (2015)

Linear relationship between ampicillin and glycoprotein concentrations and the diameter of the zone of inhibition. (A) Amp against E. coli, (B) Amp against B. subtilis, (C) relationship between the diameter of inhibition zone and ampicillin concentration and (D). glp G208 against E. coli, (E) glp G208 against B. subtilis, and (F) relationship between the diameter of inhibition zone and glp concentrations. Columns represent means and standard errors.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4499756&req=5

Figure 2: Linear relationship between ampicillin and glycoprotein concentrations and the diameter of the zone of inhibition. (A) Amp against E. coli, (B) Amp against B. subtilis, (C) relationship between the diameter of inhibition zone and ampicillin concentration and (D). glp G208 against E. coli, (E) glp G208 against B. subtilis, and (F) relationship between the diameter of inhibition zone and glp concentrations. Columns represent means and standard errors.
Mentions: In vitro evaluation of susceptibility of clinical isolates was performed using a semi-quantitative radial well-diffusion and broth microdilution assay. To compare the results of bacterial susceptibility to honey glycoproteins in a quantitative manner, the reference method has been designed that established the relationship between ampicillin dilutions and zone of inhibition (ZOI) in well-diffusion assay. As shown in Figure 2, the series of twofold diluted honey glycoproteins and ampicillin (1 mg/ml stock solutions) produced concentration-dependent ZOIs with R2 = 0.99, for ampicillin and R2 = 0.96 and 0.95 for glps tested against E. coli and B. subtilis, respectively. MIC values from well-diffusion assay against E. coli and B. subtilis (1 × 106 CFU/ml) were <0.025 μg/well or <2.0 μg/ml for ampicillin and <0.046 μg/well or 5.7 μg/ml for glps.

Bottom Line: Their resistance to different classes of antibiotics was confirmed using automated system Vitek 2.Glps isolated from different honeys showed a similar ability to overcome bacterial resistance to β-lactams suggesting that (a) their mode of action is distinct from other classes of β-lactams and that (b) the common glps structure was the lead structure responsible for the activity.The results of the current study together with our previous evidence of a rapid bactericidal activity of glps demonstrate that glps possess suitable characteristics to be considered a novel antibacterial drug candidate.

View Article: PubMed Central - PubMed

Affiliation: Department of Drug Discovery and Development Department St. Catharines, ON, Canada.

ABSTRACT
The emergence of extended- spectrum β-lactamase (ESBL) is the underlying cause of growing antibiotic resistance among Gram-negative bacteria to β-lactam antibiotics. We recently reported the discovery of honey glycoproteins (glps) that exhibited a rapid, concentration-dependent antibacterial activity against both Gram-positive Bacillus subtilis and Gram-negative Escherichia coli that resembled action of cell wall-active β-lactam drugs. Glps showed sequence identity with the Major Royal Jelly Protein 1 (MRJP1) precursor that harbors three antimicrobial peptides: Jelleins 1, 2, and 4. Here, we used semi-quantitative radial diffusion assay and broth microdilution assay to evaluate susceptibility of a number of multi-drug resistant (MDR) clinical isolates to the MRJP1-contaning honey glycoproteins. The MDR bacterial strains comprised three methicillin-resistant Staphylococcus aureus (MRSA), four Pseudomonas aeruginosa, two Klebsiella pneumoniae, two vancomycin-resistant Enterococci (VRE), and five ESBL identified as one Proteus mirabilis, three E. coli, and one E. coli NDM. Their resistance to different classes of antibiotics was confirmed using automated system Vitek 2. MDR isolates differed in their susceptibility to glps with MIC90 values ranging from 4.8 μg/ml against B. subtilis to 14.4 μg/ml against ESBL K. pneumoniae, Klebsiella spp. ESBL and E. coli and up to 33 μg/ml against highly resistant strains of P. aeruginosa. Glps isolated from different honeys showed a similar ability to overcome bacterial resistance to β-lactams suggesting that (a) their mode of action is distinct from other classes of β-lactams and that (b) the common glps structure was the lead structure responsible for the activity. The results of the current study together with our previous evidence of a rapid bactericidal activity of glps demonstrate that glps possess suitable characteristics to be considered a novel antibacterial drug candidate.

No MeSH data available.


Related in: MedlinePlus