Tumbarski Yulian, Deseva Ivelina, Mihaylova Dasha, Stoyanova Magdalena, Krastev Lutsian, Nikolova Radosveta, Yanakieva Velichka, Ivanov Ivan
Department of Microbiology, University of Food Technologies, 26, Maritsa Blvd., 4002 Plovdiv, Bulgaria.
Department of Analytical Chemistry and Physicochemistry, University of Food Technologies, 26, Maritsa Blvd., 4002 Plovdiv, Bulgaria.
Food Technol Biotechnol. 2018 Dec;56(4):546-552. doi: 10.17113/ftb.56.04.18.5905.
Members of the bacterial genus are known as producers of a broad spectrum of antibiotic compounds of proteinaceous nature that possess inhibitory activity against different saprophytic and pathogenic microorganisms. In the current research, a peptide synthesized by strain BM47, previously isolated from a natural thermal spring in Bulgaria, was identified and characterized as a bacteriocin. antimicrobial screening of the crude bacteriocin substance of BM47 showed activity against the plant pathogenic fungi , , sp., and Gram-negative bacterium . The antimicrobial activity of the crude bacteriocin substance was partially inhibited by the enzymes trypsin, Alcalase®, Savinase®, proteinase K, papain and Esperase®, while catalase was not effective. The crude bacteriocin substance was relatively pH resistant, but sensitive to the action of heat and most organic solvents and detergents tested. To obtain the active protein fractions, crude bacteriocin substance was purified by fast protein liquid chromatography (FPLC) using a strong anion exchange column. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis demonstrated that the purified bacteriocin had molecular mass of 19 578 Da. The amino acid analysis performed by high-performance liquid chromatography (HPLC) revealed that the isolated bacteriocin consisted of 17 types of amino acids, with the highest mol fraction expressed as percent of serine (29.3), valine (10.3), alanine (9.8) and tyrosine (7.1).
细菌属的成员被认为是多种蛋白质性质抗生素化合物的生产者，这些化合物对不同的腐生和致病微生物具有抑制活性。在当前的研究中，由先前从保加利亚的一个天然温泉中分离出的BM47菌株合成的一种肽被鉴定并表征为一种细菌素。对BM47粗细菌素物质的抗菌筛选显示其对植物致病真菌、、、sp.和革兰氏阴性菌具有活性。粗细菌素物质的抗菌活性被胰蛋白酶、碱性蛋白酶、风味蛋白酶、蛋白酶K、木瓜蛋白酶和复合蛋白酶部分抑制，而过氧化氢酶则无效。粗细菌素物质对pH相对耐受，但对热以及所测试的大多数有机溶剂和洗涤剂的作用敏感。为了获得活性蛋白组分，使用强阴离子交换柱通过快速蛋白质液相色谱（FPLC）对粗细菌素物质进行了纯化。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析表明，纯化后的细菌素分子量为19578 Da。通过高效液相色谱（HPLC）进行的氨基酸分析表明，分离出的细菌素由17种氨基酸组成，摩尔分数最高的是丝氨酸（29.3%）、缬氨酸（10.3%）、丙氨酸（9.8%）和酪氨酸（7.1%）。
