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characterization of functional, safety, and gut survival related characteristics of Lactobacillus strains isolated from farmhouse goat ' s milk cheeses.

Characterization of functional, safety, and gut survival related characteristics of Lactobacillus strains isolated from farmhouse goat's milk cheeses.

机构信息

Departamento de Microbiología, Facultad de Ciencias. Universidad de Granada. Campus de Fuente Nueva s/n, 18071-Granada, Spain.

出版信息

Int J Food Microbiol. 2013 May 15;163(2-3):136-45. doi: 10.1016/j.ijfoodmicro.2013.02.015. Epub 2013 Mar 5.

Abstract

A set of 80 Lactobacillus strains (36 Lactobacillus plantarum and 44 Lactobacillus paracasei) isolated from Spanish farmhouse cheeses have been studied as to their functional and safety properties and their survival under gut-related conditions. None of these 80 Lactobacillus strains were able to hydrolyse starch. A high percentage of L. plantarum and L. paracasei strains were, however, capable of hydrolysing casein (86.1% and 68.2% respectively). For the other characteristics investigated, L. plantarum strains generally had more positive responses than L. paracasei. The latter strains tested negative for most of these characteristics, with the exception of stachyose hydrolysis, which was positive in six strains of L. paracasei. A high percentage (91.7%) of L. plantarum produced haemo-dependent catalase. Phytase was present in 10 L. plantarum and in 2 L. paracasei. Most L. plantarum (83.3%) but no L. paracasei hydrolysed bile salts. All strains were completely resistant to a challenge of pH3, but many showed a loss of viability after a subsequent exposure to 0.3% oxgall; in fact, only one L. paracasei strain and 33 L. plantarum strains (91.67%) were tolerant to both stresses. L. plantarum Mb25 and L. plantarum Mb26 were the most adherent to Caco-2 cells (adherence percentages of 36 and 7% respectively). These two strains were also the most adherent to HeLa 229 cells, with 19.3 and 16.0% adhesion respectively. The Mb26 strain inhibited the adhesion of Listeria monocytogenes to Caco-2 cells when added simultaneously to Listeria and also when added 1h before the pathogen (21.0% and 51.6% adhesion inhibition, respectively). Production of H2O2 was detected in 38.9% of L. plantarum strains and in 9.1% of L. paracasei. Twelve L. plantarum and eight L. paracasei strains produced bacteriocin-like inhibitors. PCR amplifications of several plantaricin genes suggest that all the bacteriocinogenic strains may produce plantaricin E/F and some may also manufacture the plantaricin J/K. The nine L. plantarum strains assayed for antibiotic resistance were resistant to ciprofloxacin (MIC>2 μg/ml), vancomycin (MIC>16 μg/ml), and teicoplanin (MIC>16 μg/ml). Moreover, some strains showed intermediate resistance to penicillin, tetracycline, rifampicin, and levofloxacin. We conclude that farmhouse cheeses are good sources of biotechnologically relevant lactobacilli and that the L. plantarum species shows better biotechnological properties than L. paracasei. This can be deduced from the finding of a high percentage of strains of L. plantarum that exhibit remarkable functional and inhibitory properties and high abilities to survive in gut-related conditions, which can be further developed for biotechnological applications.

摘要

从西班牙农家干酪中分离出的 80 株乳杆菌(36 株植物乳杆菌和 44 株副干酪乳杆菌)被研究了其功能和安全性特性以及在肠道相关条件下的存活能力。这 80 株乳杆菌中没有一株能够水解淀粉。然而,大量的植物乳杆菌和副干酪乳杆菌菌株能够水解酪蛋白(分别为 86.1%和 68.2%)。对于其他研究的特性,植物乳杆菌菌株通常比副干酪乳杆菌菌株有更多的阳性反应。后者菌株除了对棉子糖的水解呈阳性外,对这些特性的大部分测试均呈阴性,而副干酪乳杆菌有 6 株呈阳性。高百分比(91.7%)的植物乳杆菌产生依赖血红素的过氧化氢酶。10 株植物乳杆菌和 2 株副干酪乳杆菌存在植酸酶。大多数植物乳杆菌(83.3%)但没有副干酪乳杆菌能水解胆盐。所有菌株对 pH3 的挑战完全耐受,但许多菌株在随后暴露于 0.3%牛胆盐后失去了活力;事实上,只有一株副干酪乳杆菌菌株和 33 株植物乳杆菌菌株(91.67%)耐受这两种应激。植物乳杆菌 Mb25 和植物乳杆菌 Mb26 对 Caco-2 细胞的粘附性最高(分别为 36%和 7%)。这两种菌株对 HeLa 229 细胞的粘附性也最高,分别为 19.3%和 16.0%。Mb26 菌株在添加李斯特菌的同时或在添加李斯特菌前 1 小时添加时,抑制李斯特菌与 Caco-2 细胞的粘附(分别为 21.0%和 51.6%的粘附抑制)。在 38.9%的植物乳杆菌菌株和 9.1%的副干酪乳杆菌菌株中检测到 H2O2 的产生。12 株植物乳杆菌和 8 株副干酪乳杆菌菌株产生细菌素样抑制剂。几种乳杆菌素基因的 PCR 扩增表明,所有产细菌素的菌株可能产生乳杆菌素 E/F,有些菌株也可能产生乳杆菌素 J/K。对 9 株抗药性进行检测的植物乳杆菌菌株对环丙沙星(MIC>2 μg/ml)、万古霉素(MIC>16 μg/ml)和替考拉宁(MIC>16 μg/ml)有耐药性。此外,一些菌株对青霉素、四环素、利福平、左氧氟沙星表现出中介耐药性。我们得出的结论是,农家干酪是具有生物技术相关性的乳杆菌的良好来源,植物乳杆菌物种比副干酪乳杆菌具有更好的生物技术特性。这可以从发现大量的植物乳杆菌菌株表现出显著的功能和抑制特性以及在肠道相关条件下具有高生存能力推断出来,这些特性可以进一步开发用于生物技术应用。

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