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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

MALDI-TOF mass spectrometry and SDS-PAGE of purified bacteriocin.MALDI-TOF MS analysis of the HPLC fraction and SDS-PAGE gel picture. Lane 1 is fraction from HPLC, Lane M is Precision Plus Protein™ Dual Xtra Standards (Bio-Rad, USA).
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pone.0140434.g005: MALDI-TOF mass spectrometry and SDS-PAGE of purified bacteriocin.MALDI-TOF MS analysis of the HPLC fraction and SDS-PAGE gel picture. Lane 1 is fraction from HPLC, Lane M is Precision Plus Protein™ Dual Xtra Standards (Bio-Rad, USA).

Mentions: When the HPLC sample was subjected to SDS-PAGE, only a single band was detected confirming its high purity. The purified bacteriocin showed molecular weight approximately 2.5 kDa compared to the marker used (Fig 5). The MALDI-TOF chromatogram (Fig 5) showed that the sample had a high density peak at 2706.6855Da. Therefore the possible molecular weight of the sample was approximately 2.7 kDa.


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

Goh HF, Philip K - PLoS ONE (2015)

MALDI-TOF mass spectrometry and SDS-PAGE of purified bacteriocin.MALDI-TOF MS analysis of the HPLC fraction and SDS-PAGE gel picture. Lane 1 is fraction from HPLC, Lane M is Precision Plus Protein™ Dual Xtra Standards (Bio-Rad, USA).
© Copyright Policy
Related In: Results  -  Collection

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

pone.0140434.g005: MALDI-TOF mass spectrometry and SDS-PAGE of purified bacteriocin.MALDI-TOF MS analysis of the HPLC fraction and SDS-PAGE gel picture. Lane 1 is fraction from HPLC, Lane M is Precision Plus Protein™ Dual Xtra Standards (Bio-Rad, USA).
Mentions: When the HPLC sample was subjected to SDS-PAGE, only a single band was detected confirming its high purity. The purified bacteriocin showed molecular weight approximately 2.5 kDa compared to the marker used (Fig 5). The MALDI-TOF chromatogram (Fig 5) showed that the sample had a high density peak at 2706.6855Da. Therefore the possible molecular weight of the sample was approximately 2.7 kDa.

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