Station de Recherches Laitières, Institut National de la Recherche Agronomique-CRJ, 78352 Jouy-en-Josas Cedex, France, and Departamento de Microbiología, Facultad de Biología, and Instituto de la Grasa y sus Derivados (C.S.I.C.), U.E.I. de Biotecnología de Alimentos, 41012 Seville, Spain.
Appl Environ Microbiol. 1993 May;59(5):1416-24. doi: 10.1128/aem.59.5.1416-1424.1993.
Twenty-six strains of Lactobacillus plantarum isolated from green olive fermentations were tested for cross-antagonistic activities in an agar drop diffusion test. Cell-free supernatants from four of these strains were shown to inhibit the growth of at least one of the L. plantarum indicator strains. L. plantarum LPCO10 provided the broadest spectrum of activity and was selected for further studies. The inhibitory compound from this strain was active against some gram-positive bacteria, including clostridia and propionibacteria as well as natural competitors of L. plantarum in olive fermentation brines. In contrast, no activity against gram-negative bacteria was detected. Inhibition due to the effect of organic acids, hydrogen peroxide, or bacteriophages was excluded. Since the inhibitory activity of the active supernatant was lost after treatment with various proteolytic enzymes, this substance could be classified as a bacteriocin, designated plantaricin S. Plantaricin S was also sensitive to glycolytic and lipolytic enzymes, suggesting that it was a glycolipoprotein. It exhibited a bactericidal and nonbacteriolytic mode of action against indicator cells. This bacteriocin was heat stable (60 min at 100 degrees C), active in a pH range of 3.0 to 7.0, and also stable in crude culture supernatants during storage. Ultrafiltration studies indicated that plantaricin S occurred as multimolecular aggregates and that the size of the smallest active form is between 3 and 10 kDa. In sodium dodecyl sulfate-polyacrylamide gels, plantaricin S migrated as a peptide of ca. 2.5 kDa. Maximum production of plantaricin S was obtained in a fermentor system in unregulated pH and log-phase cultures of L. plantarum LPCO10 in MRS broth plus 4% NaCl. In these culture conditions, a second bacteriocin (designated plantaricin T) was produced in late-stationary-phase cultures of L. plantarum LPCO10. On the basis of its biological activity, its sensitivity to various enzymes, and its molecular weight (lower than that of plantaricin S) as assessed in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, plantaricin T appeared different from plantaricin S. Curing experiments with L. plantarum LPCO10 resulted in the appearance of variants that no longer produced either of the two bacteriocins but that were still immune to both of them.
从绿橄榄发酵中分离出的 26 株植物乳杆菌在琼脂滴扩散试验中进行了交叉拮抗活性测试。结果表明，其中 4 株的细胞上清液能够抑制至少一种植物乳杆菌指示菌株的生长。植物乳杆菌 LPCO10 表现出最广泛的活性，因此被选为进一步研究的对象。该菌株的抑制化合物对一些革兰氏阳性菌（包括梭菌和丙酸杆菌）以及橄榄发酵卤水中植物乳杆菌的天然竞争菌具有活性。相比之下，没有发现对革兰氏阴性菌的活性。有机酸、过氧化氢或噬菌体的影响可以排除抑制作用。由于活性上清液经各种蛋白水解酶处理后失去抑制活性，因此该物质可归类为细菌素，命名为 plantaricin S。plantaricin S 还对糖酵解和脂解酶敏感，表明它是一种糖脂蛋白。它对指示细胞表现出杀菌而非溶菌作用模式。这种细菌素对热稳定（100°C 下 60 分钟），在 pH 3.0 至 7.0 的范围内活性，在储存过程中在粗培养上清液中也稳定。超滤研究表明，plantaricin S 以多分子聚集体的形式存在，最小的活性形式的大小在 3 到 10 kDa 之间。在十二烷基硫酸钠-聚丙烯酰胺凝胶中，plantaricin S 迁移为约 2.5 kDa 的肽。在 MRS 培养基中加 4%NaCl 的未调节 pH 和对数期培养的植物乳杆菌 LPCO10 的发酵罐系统中获得 plantaricin S 的最大产量。在这些培养条件下，植物乳杆菌 LPCO10 的迟滞期培养物中还产生了第二种细菌素（命名为 plantaricin T）。根据其生物学活性、对各种酶的敏感性以及在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳中评估的分子量（低于 plantaricin S），plantaricin T 似乎与 plantaricin S 不同。用植物乳杆菌 LPCO10 进行的消除实验导致出现不再产生两种细菌素的变体，但仍对两者都具有免疫力。
