Isolation of the Bacillus subtilis antimicrobial peptide subtilosin from the dairy product-derived Bacillus amyloliquefaciens
Isolation of the Bacillus subtilis antimicrobial peptide subtilosin from the dairy product-derived Bacillus amyloliquefaciens
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DOI：
10.1111/j.1365-2672.2007.03626.x
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发表时间：
2008-04-01
影响因子：
4
通讯作者：
Chikindas, M. L.
中科院分区：
文献类型：
--
作者：
Sutyak, K. E.;Wirawan, R. E.;Chikindas, M. L.
目的：从乳制品中分离得到一种抗菌蛋白(细菌素)，并对其进行鉴定。方法与结果：通过16S核糖体RNA分析，从Yogu Farm(TM)益生菌乳饮料中分离出一株未知细菌，鉴定为芽孢杆菌属(B.amyloliquefaciens)，与枯草杆菌有亲缘关系。过夜培养的无细胞上清液对单核细胞增多性李斯特菌、阴道加德纳菌和无乳链球菌临床分离株均有抑制作用。同时，几株阴道益生菌乳杆菌对CFS产生了抗性。通过酶消化证实了该化合物具有抑制活性的性质为蛋白质类。用硫酸铵沉淀法分离蛋白，柱层析进一步纯化。结论：从解淀粉芽孢杆菌中分离到的抗菌蛋白为亚硫菌素，这是以前报道的仅由枯草芽孢杆菌产生的细菌素。本研究的意义和影响：这是首次报道亚硫菌素的种内水平转移，也是从解淀粉芽孢杆菌中分离到的第一个完全鉴定的细菌素。最后，这是关于亚硫菌素抗细菌性阴道病相关病原体活性的第一份报告。
Aims: To purify and characterize an antimicrobial protein (bacteriocin) isolated from the dairy product-derived Bacillus amyloliquefaciens.Methods and Results: An unknown bacterial species cultured from the Yogu Farm (TM) probiotic dairy beverage was identified through 16S ribosomal RNA analysis as B. amyloliquefaciens, a phylogenetically close relative of Bacillus subtilis. The cell-free supernatant (CFS) of overnight cultures was active against Listeria monocytogenes and also against clinical isolates of Gardnerella vaginalis and Streptococcus agalactiae. At the same time, several isolates of vaginal probiotic Lactobacilli were resistant to the CFS. The nature of the compound causing inhibitory activity was confirmed as proteinaceous by enzymatic digestion. The protein was isolated using ammonium sulfate precipitation, and further purified via column chromatography. PCR analysis was conducted to determine relatedness to other bacteriocins produced by Bacillus spp.Conclusion: The antimicrobial protein isolated from B. amyloliquefaciens was shown to be subtilosin, a bacteriocin previously reported as produced only by B. subtilis.Significance and Impact of the Study: This is the first report of intra-species horizontal gene transfer for subtilosin and the first fully characterized bacteriocin isolated from B. amyloliquefaciens. Finally, this is the first report on subtilosin's activity against bacterial vaginosis-associated pathogens.