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从葡萄牙受保护的原产地名称奶酪中分离出的戊糖片球菌B231:其细菌素的生产及部分特性研究

Lactobacillus pentosus B231 Isolated from a Portuguese PDO Cheese: Production and Partial Characterization of Its Bacteriocin.

作者信息

Guerreiro Joana, Monteiro Vitor, Ramos Carla, Franco Bernadette Dora Gombossy de Melo, Martinez Rafael Chacon Ruiz, Todorov Svetoslav Dimitrov, Fernandes Paulo

机构信息

Escola Superior de Tecnologia e Gestão, Instituto Politécnico de Viana do Castelo, Av. do Atlântico, 4900, Viana do Castelo, Portugal.

出版信息

Probiotics Antimicrob Proteins. 2014 Jun;6(2):95-104. doi: 10.1007/s12602-014-9157-3.

Abstract

Bacteriocin B231 produced by Lactobacillus pentosus, isolated from an artisanal raw cow's milk protected designation of origin Portuguese cheese, is a small protein with an apparent relative mass of about 5 kDa and active against a large number of Listeria monocytogenes wild-type strains, Listeria ivanovii and Listeria innocua. Bacteriocin B231 production is highly dependent on the type of the culture media used for growth of Lact. pentosus B231. Replacement of glucose with maltose yielded the highest bacteriocin production from eight different carbon sources. Similar results were recorded in the presence of combination of glucose and maltose or galactose. Production of bacteriocin B231 reached maximal levels of 800 AU/ml during the stationary phase of growth of Lact. pentosus B231 in MRS broth at 30 °C. Bacteriocin B231 (in cell-free supernatant) was sensitive to treatment with trypsin and proteinase K, but not affected by the thermal treatment in range of 55-121 °C, or freezing (-20 °C). Bacteriocin production and inhibitory spectrum were evaluated. Gene encoding plantaricin S has been detected in the genomic DNA. Virulence potential and safety of Lact. pentosus B231 were assessed by PCR targeted the genes gelE, hyl, asa1, esp, cylA, efaA, ace, vanA, vanB, hdc1, hdc2, tdc and odc. The Lact. pentosus B231 strains harbored plantaricin S gene, while the occurrence of virulence, antibiotic resistance and biogenic amine genes was limited to cytolysin, hyaluronidase, aggregation substance, adhesion of collagen protein, gelatinase, tyrosine decarboxylase and vancomycin B genes.

摘要

从葡萄牙原产地保护的手工生牛奶奶酪中分离出的戊糖片球菌产生的细菌素B231是一种表观相对分子质量约为5 kDa的小蛋白,对大量单核细胞增生李斯特菌野生型菌株、伊氏李斯特菌和无害李斯特菌具有活性。细菌素B231的产生高度依赖于用于戊糖片球菌B231生长的培养基类型。用麦芽糖替代葡萄糖,在八种不同碳源中产生的细菌素产量最高。在葡萄糖与麦芽糖或半乳糖组合存在的情况下也记录到了类似结果。在30℃的MRS肉汤中,戊糖片球菌B231生长的稳定期,细菌素B231的产量达到最大水平800 AU/ml。细菌素B231(无细胞上清液)对胰蛋白酶和蛋白酶K处理敏感,但在55 - 121℃范围内的热处理或冷冻(-20℃)均不受影响。评估了细菌素的产生和抑制谱。在基因组DNA中检测到了编码植物乳杆菌素S的基因。通过针对gelE、hyl、asa1、esp、cylA、efaA、ace、vanA、vanB、hdc1、hdc2、tdc和odc基因的PCR评估了戊糖片球菌B231的毒力潜力和安全性。戊糖片球菌B231菌株含有植物乳杆菌素S基因,而毒力、抗生素抗性和生物胺基因的出现仅限于细胞溶素、透明质酸酶、聚集物质、胶原蛋白黏附、明胶酶、酪氨酸脱羧酶和万古霉素B基因。

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