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Purification and Characterization of Bacteriocin Produced by Weissella confusa A3 of Dairy Origin.

Goh HF, Philip K - PLoS ONE (2015)

Bottom Line: Reduction of activity was shown after treatment with proteinase K, trypsin and peptidase that confirmed the proteinaceous nature of the compound.Bacterial cell treated with the bacteriocin also showed significant morphological changes under transmission electron microscope.No virulence and disease related genes can be detected from the genome of the strain.

View Article: PubMed Central - PubMed

Affiliation: Microbiology Division, Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia.

ABSTRACT
A dramatic increase in bacterial resistance towards currently available antibiotics has raised worldwide concerns for public health. Therefore, antimicrobial peptides (AMPs) have emerged as a promisingly new group of therapeutic agents for managing infectious diseases. The present investigation focusses on the isolation and purification of a novel bacteriocin from an indigenous sample of cow milk and it's mode of action. The bacteriocin was isolated from Weissella confusa A3 that was isolated from the sample and was shown to have inhibitory activity towards pathogenic bacteria namely Bacillus cereus, Escherichia coli, Pseudomonas aeruginosa and Micrococcus luteus. The bacteriocin was shown to be heat stable and functioned well at low pH (2 to 6). Reduction of activity was shown after treatment with proteinase K, trypsin and peptidase that confirmed the proteinaceous nature of the compound. MALDI-TOF analysis of the sample gave a mass approximating 2.7 kDa. The membrane of the bacteria was disrupted by the bacteriocin causing SYTOX® green dye to enter the cell and bind to the bacterial DNA giving fluorescence signal. Bacterial cell treated with the bacteriocin also showed significant morphological changes under transmission electron microscope. No virulence and disease related genes can be detected from the genome of the strain.

No MeSH data available.


Related in: MedlinePlus

Effect of various concentration of bacteriocin on the growth of B. cereus at 37°C.The optical density was measured at 615 nm every hour continuously for a period of 24 hours.
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pone.0140434.g004: Effect of various concentration of bacteriocin on the growth of B. cereus at 37°C.The optical density was measured at 615 nm every hour continuously for a period of 24 hours.

Mentions: The MIC value (Table 2) of the bacteriocin was 9.25 μg/ml against both B.cereus and M. luteus. The MIC value increased to 18.5 μg/ml against E.coli and P. aeruginosa. A higher MIC of 37 μg/ml was required to inhibit L. lactis and E. faecium. After plating 10 μl of the overnight bacteriocin with the bacterial solution from the wells but without any bacterial growth after incubation, the MBC obtained for B.cereus and M. luteus was 37 μg/ml. The MBC for E.coli and P. aeruginosa was 74 μg/ml whereas L. lactis and E. faecium required a concentration exceeding 74 μg/ml to produce bactericidal effect. The addition of different concentrations of bacteriocin affected the growth of B. cereus (Fig 4). The growth of bacteria can be observed in concentrations below 4.63 μg/ml. The growth of B. cereus was extremely slow in concentrations of 9.25 and 18.5 μg/ml. The optical density of B. cereus remained almost unchanged in concentrations of bacteriocin exceeding 37 μg/ml.


Purification and Characterization of Bacteriocin Produced by Weissella confusa A3 of Dairy Origin.

Goh HF, Philip K - PLoS ONE (2015)

Effect of various concentration of bacteriocin on the growth of B. cereus at 37°C.The optical density was measured at 615 nm every hour continuously for a period of 24 hours.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4608715&req=5

pone.0140434.g004: Effect of various concentration of bacteriocin on the growth of B. cereus at 37°C.The optical density was measured at 615 nm every hour continuously for a period of 24 hours.
Mentions: The MIC value (Table 2) of the bacteriocin was 9.25 μg/ml against both B.cereus and M. luteus. The MIC value increased to 18.5 μg/ml against E.coli and P. aeruginosa. A higher MIC of 37 μg/ml was required to inhibit L. lactis and E. faecium. After plating 10 μl of the overnight bacteriocin with the bacterial solution from the wells but without any bacterial growth after incubation, the MBC obtained for B.cereus and M. luteus was 37 μg/ml. The MBC for E.coli and P. aeruginosa was 74 μg/ml whereas L. lactis and E. faecium required a concentration exceeding 74 μg/ml to produce bactericidal effect. The addition of different concentrations of bacteriocin affected the growth of B. cereus (Fig 4). The growth of bacteria can be observed in concentrations below 4.63 μg/ml. The growth of B. cereus was extremely slow in concentrations of 9.25 and 18.5 μg/ml. The optical density of B. cereus remained almost unchanged in concentrations of bacteriocin exceeding 37 μg/ml.

Bottom Line: Reduction of activity was shown after treatment with proteinase K, trypsin and peptidase that confirmed the proteinaceous nature of the compound.Bacterial cell treated with the bacteriocin also showed significant morphological changes under transmission electron microscope.No virulence and disease related genes can be detected from the genome of the strain.

View Article: PubMed Central - PubMed

Affiliation: Microbiology Division, Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia.

ABSTRACT
A dramatic increase in bacterial resistance towards currently available antibiotics has raised worldwide concerns for public health. Therefore, antimicrobial peptides (AMPs) have emerged as a promisingly new group of therapeutic agents for managing infectious diseases. The present investigation focusses on the isolation and purification of a novel bacteriocin from an indigenous sample of cow milk and it's mode of action. The bacteriocin was isolated from Weissella confusa A3 that was isolated from the sample and was shown to have inhibitory activity towards pathogenic bacteria namely Bacillus cereus, Escherichia coli, Pseudomonas aeruginosa and Micrococcus luteus. The bacteriocin was shown to be heat stable and functioned well at low pH (2 to 6). Reduction of activity was shown after treatment with proteinase K, trypsin and peptidase that confirmed the proteinaceous nature of the compound. MALDI-TOF analysis of the sample gave a mass approximating 2.7 kDa. The membrane of the bacteria was disrupted by the bacteriocin causing SYTOX® green dye to enter the cell and bind to the bacterial DNA giving fluorescence signal. Bacterial cell treated with the bacteriocin also showed significant morphological changes under transmission electron microscope. No virulence and disease related genes can be detected from the genome of the strain.

No MeSH data available.


Related in: MedlinePlus