College of Animal Science, MOE Key Laboratory of Molecular Animal Nutrition, Zhejiang University, Hangzhou 310029, PR China.
Anaerobe. 2013 Aug;22:97-103. doi: 10.1016/j.anaerobe.2013.06.005. Epub 2013 Jun 21.
Eighty-five strains of lactobacillus were isolated from the pig intestine and identified by sequencing analysis based on 16S rRNA gene, from which five lactobacillus strains with high adhesive ability were selected. The inhibition ability of the five lactobacillus strains with or without S-layer proteins against adherence of Escherichia coli K88 and Salmonella enteritidis 50335 to Caco-2 was evaluated in vitro with Lactobacillus rhamnosus GG strain (LGG) as a positive control. In addition, tolerance of lactobacilli to heat, acid, bile, Zn(2+) and Cu(2+) were assessed. All five selected strains, Lactobacillus salivarius ZJ614 (JN981856), Lactobacillus reuteri ZJ616 (JN981858), L. reuteri ZJ617 (JN981859), L. reuteri ZJ621 (JN981863) and L. reuteri ZJ623 (JN981865), showed inhibition against the two pathogens, E. coli K88 and S. enteritidis 50335. L. reuteri ZJ621 showed higher inhibition ability than the others to S. enteritidis 50335 (P < 0.05). Sodium dodecyl sulfate-Polyacrylamide gel electrophoresis (SDS-PAGE) analysis indicated that all five strains had abundant bands with molecular weight ranging from 34 to 130 KDa as well as had a common band of approximately 42 KDa. After treatment with 5 M LiCl to remove S-layer protein, the inhibition activities of the lactobacilli against pathogens decreased significantly (P < 0.05). The results showed that higher adhesive ability means higher inhibition activity for lactobacillus against pathogen, in which S-layer proteins plays an important role.
从猪肠道中分离出 85 株乳杆菌，通过 16S rRNA 基因测序分析进行鉴定，从中选择 5 株具有高黏附能力的乳杆菌菌株。以鼠李糖乳杆菌 GG 株（LGG）为阳性对照，体外评估 5 株乳杆菌菌株及其 S 层蛋白缺失株对大肠杆菌 K88 和肠炎沙门氏菌 50335 黏附 Caco-2 细胞的抑制能力。此外，评估了乳杆菌对热、酸、胆盐、Zn(2+)和 Cu(2+)的耐受性。所有 5 株选定的菌株，唾液乳杆菌 ZJ614（JN981856）、雷特氏乳杆菌 ZJ616（JN981858）、L. reuteri ZJ617（JN981859）、L. reuteri ZJ621（JN981863）和 L. reuteri ZJ623（JN981865）均表现出对两种病原体（大肠杆菌 K88 和肠炎沙门氏菌 50335）的抑制作用。L. reuteri ZJ621 对肠炎沙门氏菌 50335 的抑制能力高于其他菌株（P<0.05）。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分析表明，所有 5 株菌均具有丰富的分子量在 34 至 130 kDa 之间的条带，并且具有约 42 kDa 的共同条带。用 5 M LiCl 处理以去除 S 层蛋白后，乳杆菌对病原体的抑制活性显著降低（P<0.05）。结果表明，较高的黏附能力意味着乳杆菌对病原体的抑制活性较高，其中 S 层蛋白起着重要作用。
