Characterization of a bacteriocin produced by Prevotella nigrescens ATCC 25261.
OBJECTIVE
To characterize the antimicrobial activity produced by Prevotella nigrescens ATCC 25261, and to evaluate its safety on cultured gingival fibroblasts.


RESULTS
An antimicrobial activity was obtained from purifying the culture supernatant of Pr. nigrescens ATCC 25261. Purification of the active compound was achieved with ammonium sulphate precipitation followed by anion-exchange and gel filtration chromatography. As revealed by SDS-PAGE, the active fraction was relatively homogeneous, showing a protein with an approximate molecular weight of 41 kDa. The antimicrobial compound, named nigrescin, exhibited a bactericidal mode of action against Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythensis, and Actinomyces spp. Nigrescin was stable in a pH range between 6.5 and 9.5, at 100 degrees C for 10 min, and resistant to lyophilization. But its activity was lost after proteinase K treatment. Despite at very high concentrations beyond the minimum inhibitory concentration (MIC), nigrescin was not toxic to the gingival fibroblasts.


CONCLUSIONS
Nigrescin is a novel bacteriocin produced by Pr. nigrescens ATCC 25261. It exhibits antimicrobial activity against species that are implicated in periodontal diseases. The absence of toxicity on the gingival fibroblasts suggests the possibility in using of nigrescin for an application in periodontal treatment.


CONCLUSIONS
Novel evidence on nigrescin would make Pr. nigrescens ATCC 25261 attractive in biotechnological applications as an antimicrobial agent in clinical dentistry.
Kaewsrichan
J
J
Department of Stomatology, Faculty of Dentistry, Prince of Songkla University, Hat-Yai, Songkhla 90112, Thailand. mjasdee@ratree.psu.ac.th
Douglas
C W I
CW
Nissen-Meyer
J
J
Fimland
G
G
Teanpaisan
R
R
D000190:Actinomyces / Q000187:drug effects; D000645:Ammonium Sulfate; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology / Q000633:toxicity; D001430:Bacteriocins / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology* / Q000633:toxicity; D001439:Bacteroides / Q000187:drug effects; D002478:Cells, Cultured; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D019286:Endopeptidase K / Q000378:metabolism; D004795:Enzyme Stability; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects; D005590:Fractional Precipitation; D005612:Freeze Drying; D005881:Gingiva / Q000166:cytology / Q000187:drug effects; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D016966:Porphyromonas gingivalis / Q000187:drug effects; D018713:Prevotella intermedia / Q000187:drug effects; D045242:Prevotella nigrescens / Q000378:metabolism*; D013696:Temperature
D000900:Anti-Bacterial Agents; D001430:Bacteriocins; D019286:Endopeptidase K; D000645:Ammonium Sulfate