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鉴定和评价植物乳杆菌 FT259 的益生菌潜力,该菌株是从巴西半硬质传统奶酪中分离出的具有细菌素产生能力的菌株。

Identification and evaluation of the probiotic potential of Lactobacillus paraplantarum FT259, a bacteriocinogenic strain isolated from Brazilian semi-hard artisanal cheese.

机构信息

Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo (FCFRP-USP), Avenida do Café s/n, 14040-903 Ribeirão Preto, São Paulo, Brazil.

出版信息

Anaerobe. 2013 Aug;22:57-63. doi: 10.1016/j.anaerobe.2013.06.006. Epub 2013 Jun 20.

Abstract

This study aimed to identify a bacteriocinogenic Lactobacillus isolate (FT259) obtained from Brazilian semi-hard Minas type cheese and to evaluate its probiotic and antimicrobial potentials. The strain was identified by biochemical tests (at genus level), and by 16S rDNA sequencing combined with recA gene amplification (for species). To determine the inhibitory spectrum towards food borne pathogens and lactic acid bacteria, the spot-on-the-lawn assay was carried out. Moreover, the proteinaceous nature of the antimicrobial compound produced was evaluated by susceptibility to degradation by proteolytic enzymes. The isolated strain was tested for survival in acidified culture media (pH 2.0, 2.5 and 3.5), in vitro tolerance to bile salts and viability under gastric conditions. Adhesion of Lactobacillus paraplantarum FT259 to Caco-2 cells was evaluated by surface plate count on De Man, Rogosa, and Sharpe (MRS) agar and also by FISH method (fluorescent in situ hybridization) with the aid of Eub338 probe for fluorescence microscopy analysis. The isolate was identified as L. paraplantarum FT259 and it produced bacteriocins that inhibited the growth of Listeria monocytogenes, Listeria innocua and several lactic acid bacteria. It was also observed that L. paraplantarum FT259 tolerated exposure to pH 3.5, and bile salts 0.3% for up to 180 min. In experiments with simulated gastric juice, viable cells of L. paraplantarum FT259 decreased from 8.6 log CFU/mL to 3.5 log CFU/mL after 180 min. For the same strain, in studies with Caco-2 cells, 74% of adhesion was observed through plate count and FISH assays. It was also demonstrated isolated FT259 was susceptible to the majority the antibiotics tested. Overall, the results indicated L. paraplantarum FT259 is a potential probiotic and the production of bacteriocin may be an interesting feature for food applications.

摘要

本研究旨在从巴西半硬型 Minas 奶酪中分离出一株具有抑菌活性的乳酸菌(FT259),并评估其益生菌和抗菌潜力。该菌株通过生化试验(属水平)和 16S rDNA 测序结合 recA 基因扩增(种水平)进行鉴定。为了确定其对食源性病原体和乳酸菌的抑菌谱,采用点植法进行检测。此外,还通过蛋白酶降解试验评估了抗菌化合物的蛋白性质。该分离株在酸化培养基(pH 2.0、2.5 和 3.5)中的生存能力、胆汁盐的体外耐受性以及在胃条件下的生存能力进行了测试。采用表面平板计数法(MRS 琼脂)和荧光原位杂交(FISH)法(使用 Eub338 探针进行荧光显微镜分析)评估 Lactobacillus paraplantarum FT259 对 Caco-2 细胞的黏附能力。该分离株被鉴定为 L. paraplantarum FT259,它产生的细菌素可以抑制单核细胞增生李斯特菌、无害李斯特菌和几种乳酸菌的生长。还观察到 L. paraplantarum FT259 可以耐受 pH 3.5 和 0.3%的胆汁盐,最长可达 180 分钟。在模拟胃液实验中,L. paraplantarum FT259 的活菌数从 8.6logCFU/mL 减少到 3.5logCFU/mL ,180 分钟后。对于同一菌株,在 Caco-2 细胞研究中,平板计数和 FISH 检测均观察到 74%的黏附。还证明分离的 FT259 对大多数测试的抗生素敏感。总的来说,结果表明 L. paraplantarum FT259 是一种有潜力的益生菌,其细菌素的产生可能是食品应用的一个有趣特征。

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