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Assessing the antimicrobial activities of Ocins.

Choyam S, Lokesh D, Kempaiah BB, Kammara R - Front Microbiol (2015)

Bottom Line: The generation of a zone of inhibition on a solid substrate indicates the bioactivity of antimicrobial peptides such as bacteriocin and enterocin.However, universal well diffusion assays for antimicrobials, irrespective of their ability to diffuse (bacteriocin and enterocin), do not exist.The ability of different zymography components to generate non-specific activities have rarely been explored in the literature.

View Article: PubMed Central - PubMed

Affiliation: Department of Protein Chemistry and Technology, Central Food Technological Research Institute (CFTRI) Mysore, India.

ABSTRACT
The generation of a zone of inhibition on a solid substrate indicates the bioactivity of antimicrobial peptides such as bacteriocin and enterocin. The indicator strain plays a significant role in bacteriocin assays. Other characteristics of bacteriocins, such as their dispersal ability and the different zymogram components, also affect bacteriocin assays. However, universal well diffusion assays for antimicrobials, irrespective of their ability to diffuse (bacteriocin and enterocin), do not exist. The ability of different zymography components to generate non-specific activities have rarely been explored in the literature. The purpose of the present work was to evaluate the impact of major factors (diffusion and rate of diffusion) in a solid substrate bioassay, and to document the adverse effects of sodium dodecyl sulfate in zymograms used to estimate the approximate molecular weight of bacteriocins.

No MeSH data available.


Related in: MedlinePlus

(A–D) Bacteriocin and enterocin inhibition of a Micrococcus luteus indicator lawn on de Man, Rogosa, and Sharpe (MRS) agar medium. The producer strains are Bifiodobacterium catenulatum and Enteroccus gallinarum.(A) Wells 1 and 2 are unique wells, and they share the same structure as shown in (B). Wells 3 and 4 are conventional wells made by a gel borer, and they share the same structure as shown in (B). Numbers 5 and 6 are sterile disks placed on the lawn. Wells 1 and 3 and disk 5 correspond to enterocin, and wells 2 and 4, and disk 6 correspond to bacteriocin. (D) Nisin subjected to three different kinds of assays.
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Figure 1: (A–D) Bacteriocin and enterocin inhibition of a Micrococcus luteus indicator lawn on de Man, Rogosa, and Sharpe (MRS) agar medium. The producer strains are Bifiodobacterium catenulatum and Enteroccus gallinarum.(A) Wells 1 and 2 are unique wells, and they share the same structure as shown in (B). Wells 3 and 4 are conventional wells made by a gel borer, and they share the same structure as shown in (B). Numbers 5 and 6 are sterile disks placed on the lawn. Wells 1 and 3 and disk 5 correspond to enterocin, and wells 2 and 4, and disk 6 correspond to bacteriocin. (D) Nisin subjected to three different kinds of assays.

Mentions: The standardized test was repeated without making any wells on the plate, but by placing sterile disks (Hi-Media laboratories, Mumbai, India; 5–10 mm diameter) on the agar surface (Figure 1A, wells 5 and 6). The bacteriocin/enterocin-saturated disks were used in the well diffusion assays (Figure 1A, wells 1 and 3; Figure 1B well a; and Figure 1C) after drying the plates for 30 min to allow the absorption and distribution of the bacteriocin into the medium. Eventually, the plates were dried and then incubated at 37°C for 24 h or until ZOI developed to compare the bacteriocin and enterocin activities under different conditions. The diameters of the ZOI in the assay plates were measured using images of the plates and graded scales. The tests were performed in triplicate.


Assessing the antimicrobial activities of Ocins.

Choyam S, Lokesh D, Kempaiah BB, Kammara R - Front Microbiol (2015)

(A–D) Bacteriocin and enterocin inhibition of a Micrococcus luteus indicator lawn on de Man, Rogosa, and Sharpe (MRS) agar medium. The producer strains are Bifiodobacterium catenulatum and Enteroccus gallinarum.(A) Wells 1 and 2 are unique wells, and they share the same structure as shown in (B). Wells 3 and 4 are conventional wells made by a gel borer, and they share the same structure as shown in (B). Numbers 5 and 6 are sterile disks placed on the lawn. Wells 1 and 3 and disk 5 correspond to enterocin, and wells 2 and 4, and disk 6 correspond to bacteriocin. (D) Nisin subjected to three different kinds of assays.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4585010&req=5

Figure 1: (A–D) Bacteriocin and enterocin inhibition of a Micrococcus luteus indicator lawn on de Man, Rogosa, and Sharpe (MRS) agar medium. The producer strains are Bifiodobacterium catenulatum and Enteroccus gallinarum.(A) Wells 1 and 2 are unique wells, and they share the same structure as shown in (B). Wells 3 and 4 are conventional wells made by a gel borer, and they share the same structure as shown in (B). Numbers 5 and 6 are sterile disks placed on the lawn. Wells 1 and 3 and disk 5 correspond to enterocin, and wells 2 and 4, and disk 6 correspond to bacteriocin. (D) Nisin subjected to three different kinds of assays.
Mentions: The standardized test was repeated without making any wells on the plate, but by placing sterile disks (Hi-Media laboratories, Mumbai, India; 5–10 mm diameter) on the agar surface (Figure 1A, wells 5 and 6). The bacteriocin/enterocin-saturated disks were used in the well diffusion assays (Figure 1A, wells 1 and 3; Figure 1B well a; and Figure 1C) after drying the plates for 30 min to allow the absorption and distribution of the bacteriocin into the medium. Eventually, the plates were dried and then incubated at 37°C for 24 h or until ZOI developed to compare the bacteriocin and enterocin activities under different conditions. The diameters of the ZOI in the assay plates were measured using images of the plates and graded scales. The tests were performed in triplicate.

Bottom Line: The generation of a zone of inhibition on a solid substrate indicates the bioactivity of antimicrobial peptides such as bacteriocin and enterocin.However, universal well diffusion assays for antimicrobials, irrespective of their ability to diffuse (bacteriocin and enterocin), do not exist.The ability of different zymography components to generate non-specific activities have rarely been explored in the literature.

View Article: PubMed Central - PubMed

Affiliation: Department of Protein Chemistry and Technology, Central Food Technological Research Institute (CFTRI) Mysore, India.

ABSTRACT
The generation of a zone of inhibition on a solid substrate indicates the bioactivity of antimicrobial peptides such as bacteriocin and enterocin. The indicator strain plays a significant role in bacteriocin assays. Other characteristics of bacteriocins, such as their dispersal ability and the different zymogram components, also affect bacteriocin assays. However, universal well diffusion assays for antimicrobials, irrespective of their ability to diffuse (bacteriocin and enterocin), do not exist. The ability of different zymography components to generate non-specific activities have rarely been explored in the literature. The purpose of the present work was to evaluate the impact of major factors (diffusion and rate of diffusion) in a solid substrate bioassay, and to document the adverse effects of sodium dodecyl sulfate in zymograms used to estimate the approximate molecular weight of bacteriocins.

No MeSH data available.


Related in: MedlinePlus