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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

Antimicrobial assay.Antimicrobial activity of different fractions from Amberlite XAD 16 column (a) and two different fractions (< 2kDa and 2–5kDa) from Vivaspin (b).
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pone.0140434.g002: Antimicrobial assay.Antimicrobial activity of different fractions from Amberlite XAD 16 column (a) and two different fractions (< 2kDa and 2–5kDa) from Vivaspin (b).

Mentions: The bacteriocin produced by the W. confusa showed inhibition against all test bacteria except L. monocytogenes NCTC10890 and S. aureus RF122. The inhibition zone of the ammonium sulphate precipitated extract against different test bacteria is shown in Table 2. The crude bacteriocin was desalted with XAD 16 column to remove the ammonium sulphate. Three fractions were obtained from the Amberlite XAD 16 column. Only fractions eluted out with 50% and 90% of acetonitrile gave activity against B. cereus (Fig 2a). The active fractions were combined and fractionated with Vivaspin column. The sample fraction of size 2–5kDa showed positive result when tested against B. cereus (Fig 2b). The bacteriocin from Vivaspin was injected into HPLC and the run under the set gradient for 1 hour (Fig 3). The active bacteriocin was eluted out from 36 to 39 min and inhibited B. cereus.


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

Goh HF, Philip K - PLoS ONE (2015)

Antimicrobial assay.Antimicrobial activity of different fractions from Amberlite XAD 16 column (a) and two different fractions (< 2kDa and 2–5kDa) from Vivaspin (b).
© Copyright Policy
Related In: Results  -  Collection

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

pone.0140434.g002: Antimicrobial assay.Antimicrobial activity of different fractions from Amberlite XAD 16 column (a) and two different fractions (< 2kDa and 2–5kDa) from Vivaspin (b).
Mentions: The bacteriocin produced by the W. confusa showed inhibition against all test bacteria except L. monocytogenes NCTC10890 and S. aureus RF122. The inhibition zone of the ammonium sulphate precipitated extract against different test bacteria is shown in Table 2. The crude bacteriocin was desalted with XAD 16 column to remove the ammonium sulphate. Three fractions were obtained from the Amberlite XAD 16 column. Only fractions eluted out with 50% and 90% of acetonitrile gave activity against B. cereus (Fig 2a). The active fractions were combined and fractionated with Vivaspin column. The sample fraction of size 2–5kDa showed positive result when tested against B. cereus (Fig 2b). The bacteriocin from Vivaspin was injected into HPLC and the run under the set gradient for 1 hour (Fig 3). The active bacteriocin was eluted out from 36 to 39 min and inhibited B. cereus.

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