Antibacterial activity of a bacteriocin from Lactobacillus paracasei HL32 against Porphyromonas gingivalis.
Porphyromonas gingivalis infections cause problems in periodontal diseases and in certain systemic diseases. There is evidence that Lactobacillus spp. can control populations of P. gingivalis, but there are few data on the effects of purified bacteriocins from Lactobacillus paracasei HL32 on P. gingivalis. The objective of this study was to examine the antibacterial activity of a bacteriocin from L. paracasei HL32 and to relate this activity to its composition. A bacteriocin was purified from culture supernatants of Lactobacillus spp. using a dialysis technique followed by gel-permeation chromatography. Composition of the bacteriocin was characterised by ninhydrin tests, ultraviolet spectrophotometry, thin-layer chromatography, sodium-dodecyl sulphate-polyacrylamide gel electrophoresis, electrospray ionisation mass spectrometry and amino acid analysis. The amino acid sequence from the N-terminal of the bacteriocin was determined. Antibacterial activity was examined by the cylinder plate method, microtitre assay and scanning electron microscopy as compared with standard antibiotics. The bacteriocin had a molecular weight of approximately 56kDa, was comprised of 68% carbohydrate and 32% protein, and showed maximum peak absorbance at 214 and 254nm. The bacteriocin was found to be effective against P. gingivalis; it caused swelling and pore formation on the cell envelope at a minimum bactericidal concentration of 0.14mM, and caused death within 2h. Metronidazole killed P. gingivalis but did not affect the envelope, whereas tetracycline affected P. gingivalis with cell deformation. In conclusion, the bacteriocin from L. paracasei HL32 had the ability to kill P. gingivalis, suggesting that it could be a promising alternative chemotherapeutic agent for P. gingivalis infections.
Pangsomboon
Kanokporn
K
Prince of Songkla University, Hat Yai, Songkla, Thailand.
Kaewnopparat
Sanae
S
Pitakpornpreecha
Thanawat
T
Srichana
Teerapol
T
D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000302:isolation & purification / Q000627:therapeutic use*; D001430:Bacteriocins / Q000235:genetics / Q000302:isolation & purification / Q000627:therapeutic use*; D016866:Bacteroidaceae Infections / Q000188:drug therapy*; D002473:Cell Wall / Q000187:drug effects; D002855:Chromatography, Thin Layer; D004591:Electrophoresis, Polyacrylamide Gel; D007778:Lactobacillus / Q000378:metabolism*; D008826:Microbial Sensitivity Tests; D008855:Microscopy, Electron, Scanning; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010518:Periodontitis / Q000188:drug therapy*; D016966:Porphyromonas gingivalis / Q000187:drug effects* / Q000648:ultrastructure; D021241:Spectrometry, Mass, Electrospray Ionization
D000900:Anti-Bacterial Agents; D001430:Bacteriocins