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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

Membrane permeability test.Real time PCR fluorescence pattern of bacteriocin from W. confusa, negative control, positive control (NaOH, 1M) and tetracycline.
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pone.0140434.g006: Membrane permeability test.Real time PCR fluorescence pattern of bacteriocin from W. confusa, negative control, positive control (NaOH, 1M) and tetracycline.

Mentions: The real time PCR fluorescence (Fig 6) showed that the bacteria treated with bacteriocin from W. confusa had highest fluorescence density after positive control (NaOH, 1M). Negative controls without adding any bacteriocin and with tetracycline added showed the lowest fluorescence. This result proved membrane disruption occur when bacteriocin from W. confusa was added into the test bacteria allowed the fluorescence dye to bind to the nucleic acid of the test bacteria and hence giving high fluorescence intensity.


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

Goh HF, Philip K - PLoS ONE (2015)

Membrane permeability test.Real time PCR fluorescence pattern of bacteriocin from W. confusa, negative control, positive control (NaOH, 1M) and tetracycline.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0140434.g006: Membrane permeability test.Real time PCR fluorescence pattern of bacteriocin from W. confusa, negative control, positive control (NaOH, 1M) and tetracycline.
Mentions: The real time PCR fluorescence (Fig 6) showed that the bacteria treated with bacteriocin from W. confusa had highest fluorescence density after positive control (NaOH, 1M). Negative controls without adding any bacteriocin and with tetracycline added showed the lowest fluorescence. This result proved membrane disruption occur when bacteriocin from W. confusa was added into the test bacteria allowed the fluorescence dye to bind to the nucleic acid of the test bacteria and hence giving high fluorescence intensity.

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