Bile salt hydrolase and antimicrobial activities of three bile resistant probiotics Lactobacillus plantarum strains isolated from Cameroonian artisanal fermented milk

Bemmo Kamdem Ulrich Landry, Kaktcham Pierre-Marie, Momo Kenfack Chancel Hector, Foko Kouam Edith Marius, Z. François, Wang Yan, Zhu Taicheng, Yin Li
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引用次数: 2

Abstract

Probiotics are well known for their efficacy as dietary adjuncts providing benefits to consumers. However, the selection of probiotics before incorporation into diet requires the scrutiny of a well-defined set of criteria. Hence, three Lactobacillus plantarum strains (GGU, GLA51 and GLP56, having KU949009, KU949010 and KU949011 respectively as GenBank accession numbers) previously isolated from Cameroonian traditional fermented milk have been used in this study for the evaluation of their Bile salt hydrolase and antimicrobial activities. Bile tolerance test was carried out by monitoring the bacterial growth at different Oxgall bile concentration (0.3%, 0.5% and 1%). The bile salt hydrolase activity was measured by determining the amount of amino acid liberated from conjugated bile salts by lactobacilli strains. Also, three bsh genes (bsh1 gene specific to Lactobacillus plantarum species, bshA and bshB genes specific to Lactobacillus acidophilus species) involved in the bile salt hydrolase activity were screened. The a ntimicrobial activity of the strains was evaluated using the agar-spot test. T he three Lactobacillus plantarum strains showed survival percentages higher than 90% in the presence of 0.3% of Oxgall bile, and the delays of growth ranged from 0 to 10 min. Their bile salt hydrolase activity ranged from 15.62 ± 3.00 to 23.91 ± 5.82 U/mg towards Oxgall and from 10.47 ± 2.76 to 24.57 ± 6.31 U/mg towards Taurodeoxycholate. Only Lactobacillus plantarum GLA51 was found to harbor thebsh1 gene. Its sequence was deposited in the Genbank under the accession number MF098542. The analysis of thisbsh1 gene sequence from L. plantarum GLA51 indicated that it contained an open reading frame (ORF) of 838 nucleotides encoding a 278 amino acids protein. The three Lactobacilli strains showed inhibitory activity against the pathogenic or spoilage bacteria tested, except for Shigella flexneri. These results suggest that the three Lactobacillus plantarum strain show potential for probiotic applications. Keywords: Bile salt hydrolase activity, antimicrobial activity, Lactobacillus plantarum , bile tolerance
从喀麦隆手工发酵乳中分离的三种抗胆汁益生菌植物乳杆菌的胆汁盐水解酶和抗菌活性
益生菌是众所周知的功效,作为膳食补充剂,为消费者提供好处。然而,在将益生菌纳入饮食之前,益生菌的选择需要仔细审查一套定义良好的标准。因此,本研究利用此前从喀麦隆传统发酵乳中分离到的三株植物乳杆菌(GGU、GLA51和GLP56,分别以KU949009、KU949010和KU949011为GenBank登录号),对其胆汁盐水解酶和抗菌活性进行了评价。通过监测不同牛胆浓度(0.3%、0.5%和1%)下细菌的生长情况,进行胆汁耐受性试验。通过测定乳酸杆菌菌株结合胆汁盐释放的氨基酸量来测定胆汁盐水解酶的活性。同时,筛选到3个与胆盐水解酶活性相关的bsh基因(植物乳杆菌种特异性bsh1基因、嗜酸乳杆菌种特异性bshA和bshB基因)。采用琼脂斑点法测定菌株的抑菌活性。结果表明,3株植物乳杆菌在0.3%的牛胆中存活率均高于90%,生长延迟时间为0 ~ 10 min,对牛胆的胆盐水解酶活性为15.62±3.00 ~ 23.91±5.82 U/mg,对牛磺酸去氧胆酸的胆盐水解酶活性为10.47±2.76 ~ 24.57±6.31 U/mg。只有植物乳杆菌GLA51被发现含有bsh1基因。其序列已存入Genbank,登录号为MF098542。对植物L. plantarum GLA51中bsh1基因序列的分析表明,该基因包含一个838个核苷酸的开放阅读框(ORF),编码278个氨基酸的蛋白。除福氏志贺氏菌外,3株乳酸菌对病原菌和腐败菌均有抑制作用。这些结果表明,三种植物乳杆菌菌株具有潜在的益生菌应用潜力。关键词:胆汁盐水解酶活性;抗菌活性;植物乳杆菌
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