A model assay to demonstrate how intrinsic factors affect diffusion of bacteriocins

Blom, Henning; Katla, Tone; Hagen, Beate F; Axelsson, Lars

doi:10.1016/s0168-1605(97)00098-6

Cited by 48 publications

(37 citation statements)

References 15 publications

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“…Such guidelines should reflect the concentrations of bacteriocins used or produced in food products when bacteriocins or bacteriocinogenic strains are added as biopreservatives. Intrinsic factors affect the activity of a bacteriocin in a food product (2,5,27), but studies have shown that susceptibility differences measured in microtiter plate assays are comparable to differences seen for the same strains of L. monocytogenes in food model systems (37). Based on this study, sakacin P appears to be a candidate for inhibition of L. monocytogenes in food.…”

Section: Discussionmentioning

confidence: 74%

“…The complete purification of three class IIa bacteriocins to the amounts required for all IC 50 tests was considered a laborious task. It was therefore decided to use heterologously expressed bacteriocins from the same host strain, a system already available and used (4,5). Since the background would be constant, this was considered far better than using different (wild-type) production strains with different and unknown backgrounds.…”

Section: Discussionmentioning

confidence: 99%

“…Intrinsic properties of food have a significant influence on the effect of bacteriocins on the target cells (2,5,27,39). However, knowledge about the susceptibility of the target organism is also necessary when bacteriocins are added to food as biopreservatives.…”

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confidence: 99%

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Differences in Susceptibility of Listeria monocytogenes Strains to Sakacin P, Sakacin A, Pediocin PA-1, and Nisin

Katla

Naterstad²,

Vancanneyt

et al. 2003

Appl Environ Microbiol

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Two hundred strains of Listeria monocytogenes collected from food and the food industry were analyzed for susceptibility to the class IIa bacteriocins sakacin P, sakacin A, and pediocin PA-1 and the class I bacteriocin nisin. The individual 50% inhibitory concentrations (IC 50 ) were determined in a microtiter assay and expressed in nanograms per milliliter. The IC 50 of sakacin P ranged from 0.01 to 0.61 ng ml ؊1 . The corresponding values for pediocin PA-1, sakacin A, and nisin were 0.10 to 7.34, 0.16 to 44.2, and 2.2 to 781 ng ml ؊1 , respectively. The use of a large number of strains and the accuracy of the IC 50 determination revealed patterns not previously described, and for the first time it was shown that the IC 50 of sakacin P divided the L. monocytogenes strains into two distinct groups. Ten strains from each group were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of whole-cell proteins and amplified fragment length polymorphism. The results from these studies essentially confirmed the grouping based on the IC 50 of sakacin P. A high correlation was found between the IC 50 of sakacin P and that of pediocin PA-1 for the 200 strains. Surprisingly, the correlation between the IC 50 of the two class IIa bacteriocins sakacin A and sakacin P was lower than the correlation between the IC 50 of sakacin A and the class I bacteriocin nisin.

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Section: Discussionmentioning

confidence: 74%

Section: Discussionmentioning

confidence: 99%

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Differences in Susceptibility of Listeria monocytogenes Strains to Sakacin P, Sakacin A, Pediocin PA-1, and Nisin

Katla

Naterstad²,

Vancanneyt

et al. 2003

Appl Environ Microbiol

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“…Although the importance of these biologically based methods in the field of bacteriocins is undeniable, they also possess some major drawbacks, such as lack of specificity (35) and low reproducibility (3). The generation of antibodies to bacteriocins has provided specific and sensitive methods for the latter's detection and quantification by enabling the use of immunochemical assays (24,(26)(27)(28)35).…”

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confidence: 99%

Immunodetection of the Bacteriocin Lacticin RM: Analysis of the Influence of Temperature and Tween 80 on Its Expression and Activity

Keren

Yarmus

Halevy

et al. 2004

Appl Environ Microbiol

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Immunoassays with specific antibodies offer higher sensitivity than do bioassays with indicator strains in the detection and quantification of several bacteriocins. Here we present the purification of lacticin RM and the production of specific polyclonal antibodies to a synthetic peptide resembling an internal fragment of the mature bacteriocin. The specificity and sensitivity of the generated polyclonal antibodies were evaluated in various immunoassays. The detection limits of lacticin RM were found to be 1.9, 0.16, and 0.18 g ml ؊1 for Western blot, immuno-dot blot, and noncompetitive indirect enzyme-linked immunosorbent assays, respectively. Immunoassay sensitivities were 12.5-fold higher than that of the agar diffusion test (ADT). The production of lacticin RM showed temperature dependency, with 3, 4.2, 12.7, 28.9, 37.8, and 12 g ml ؊1 at 37, 30, 20, 15, 10, and 4°C, respectively. Temperature-stability analysis demonstrated that lacticin RM is sensitive to mild temperature, but the loss of activity does not seem to result from protein degradation. Tween 80 increased the concentration of lacticin RM eightfold and probably affected the results of the ADT either by enhancing the activity of lacticin RM or by increasing the sensitivity of the indicator strain. The use of antibodies for the specific detection and quantification of lacticin RM can expand our knowledge of its production and stability, with important implications for further investigation and future application.Bacteriocins are antibacterial proteins produced by bacteria that kill or inhibit the growth of closely related bacteria. Many lactic acid bacteria (LAB) produce a high diversity of bacteriocins, with a fairly broad inhibitory spectrum (19). In recent years, many bacteriocins from LAB belonging to different groups have been described, characterized, and purified. Nisin, produced by several strains of Lactococcus lactis, is the most studied bacteriocin, and it is used as a commercial food preservative (6,11,16). The role of LAB and their bacteriocins as food biopreservatives is expected to grow in the future as a result of consumer awareness of the potential risks derived from food-borne pathogens as well as from the artificial chemical preservatives currently used to control them (14).Lacticin RM is a bacteriocin produced by Lactococcus lactis subsp. lactis EZ26 (DSM ID-95-131), isolated from goat's milk. It displays a wide range of inhibition, including Listeria monocytogenes and Staphylococcus aureus (36). The operon encoding lacticin RM is located on a 2.5-kb fragment in the plasmid pHU1, and it is rather simple in comparison to other bacteriocin systems, including only four genes. lacA is the bacteriocin structural gene, while lacF, lacG, and lacI participate in immunity against lacticin RM (36). Preliminary results indicate that lacticin RM is optimally produced at low growth temperatures, with maximal inhibition zones against an indicator strain at 10 and 15°C. Although most of the characterized bacteriocins are produced at the producer...

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“…The effectiveness of bacteriocins to control growth of L. monocytogenes in vacuum packed cold smoked salmon has also been demonstrated by several researchers. Among them, Sakacin P has been found to be very potent against L. monocytogenes and is one of the most extensively studied bacteriocins [126][127][128][129][130][131]. Leroi et al [132] succeeded in increasing the sensory use-by-date of CSS slices by inoculating them with strains of Carnobacterium sp.…”

Section: Application Of Lab In Seafoodmentioning

confidence: 99%

Lactic Acid Bacteria and Their Bacteriocins: A Promising Approach to Seafood Biopreservation

Ghanbari¹,

Jami²

2013

Lactic Acid Bacteria - R &Amp; D for Food, Health and Livestock Purposes

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A model assay to demonstrate how intrinsic factors affect diffusion of bacteriocins

Cited by 48 publications

References 15 publications

Differences in Susceptibility of Listeria monocytogenes Strains to Sakacin P, Sakacin A, Pediocin PA-1, and Nisin

Differences in Susceptibility of Listeria monocytogenes Strains to Sakacin P, Sakacin A, Pediocin PA-1, and Nisin

Immunodetection of the Bacteriocin Lacticin RM: Analysis of the Influence of Temperature and Tween 80 on Its Expression and Activity

Lactic Acid Bacteria and Their Bacteriocins: A Promising Approach to Seafood Biopreservation

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