Pangsomboon Kanokporn, Kaewnopparat Sanae, Pitakpornpreecha Thanawat, Srichana Teerapol
Prince of Songkla University, Hat Yai, Songkla, Thailand.
Arch Oral Biol. 2006 Sep;51(9):784-93. doi: 10.1016/j.archoralbio.2006.03.008. Epub 2006 Jul 25.
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.
牙龈卟啉单胞菌感染会引发牙周疾病及某些全身性疾病。有证据表明,乳酸杆菌属可控制牙龈卟啉单胞菌的数量,但关于副干酪乳杆菌HL32纯化细菌素对牙龈卟啉单胞菌的影响的数据却很少。本研究的目的是检测副干酪乳杆菌HL32细菌素的抗菌活性,并将该活性与其组成相关联。采用透析技术继以凝胶渗透色谱法从乳酸杆菌属的培养上清液中纯化出一种细菌素。通过茚三酮试验、紫外分光光度法、薄层色谱法、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、电喷雾电离质谱法及氨基酸分析对该细菌素的组成进行了表征。测定了该细菌素N端的氨基酸序列。与标准抗生素相比,采用试管平板法、微量滴定法及扫描电子显微镜检测了抗菌活性。该细菌素的分子量约为56kDa,由68%的碳水化合物和32%的蛋白质组成,在214和254nm处显示出最大峰值吸光度。发现该细菌素对牙龈卟啉单胞菌有效;在最低杀菌浓度为0.14mM时,它会导致细胞包膜肿胀和形成孔洞,并在2小时内导致死亡。甲硝唑可杀死牙龈卟啉单胞菌,但不影响其包膜,而四环素会使牙龈卟啉单胞菌细胞变形。总之,副干酪乳杆菌HL32的细菌素具有杀死牙龈卟啉单胞菌的能力,这表明它可能是治疗牙龈卟啉单胞菌感染的一种有前景的替代化疗药物。