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使用 Biacore 分析技术筛选对人结肠黏液中唾液酸和硫酸酯残基具有黏附特性的益生菌的新方法。

New screening methods for probiotics with adhesion properties to sialic acid and sulphate residues in human colonic mucin using the Biacore assay.

机构信息

Laboratory of Animal Products Chemistry, Graduate School of Agricultural Science, Tohoku University, Sendai, Japan.

出版信息

J Appl Microbiol. 2013 Mar;114(3):854-60. doi: 10.1111/jam.12063. Epub 2013 Jan 7.

Abstract

AIMS

To determine the relationship between adhesive ability of probiotics and acidic residues in human colonic mucin, we developed a new screening method using Biacore to evaluate adherence of bacteria before and after sialic acid or sulphate residues were blocked or removed from mucin.

METHODS AND RESULTS

Ten strains of lactobacilli and three strains of bifidobacteria isolated from human faeces were evaluated for their adhesive properties to soluble human colonic mucin (sHCM) using the Biacore binding assay. Three strains (Lactobacillus strain ME-522, Lact. gasseri ME-527 and Bifidobacterium bifidum MCC1092) showing significant adherence were selected. Decreased binding activities were observed after removing sialic acid of sHCM using sialidase. However, after removing the sulphate residue using sulphatase, the adhesion of ME-527 decreased; whereas the remaining two strains had increased adhesion. The adhesion of three probiotics significantly decreased after the sulphate residue was blocked by elution with barium chloride.

CONCLUSIONS

A new evaluation method using the Biacore assay was developed to observe binding properties to the acidic residues of sHCM. Results indicated that there was a strong relationship between probiotic adhesion and acidic residues of sHCM.

SIGNIFICANCE AND IMPACT OF THE STUDY

This is the first report showing a screening method that quantitatively measures the binding between bacteria and acidic residues in sHCM using the Biacore binding assay; and provides a new method for the selection of probiotics in the future.

摘要

目的

为了确定益生菌的黏附能力与人类结肠黏液中酸性残基之间的关系,我们开发了一种新的筛选方法,使用 Biacore 评估细菌在唾液酸或硫酸盐残基被阻断或从黏蛋白中去除前后对黏蛋白的黏附能力。

方法和结果

使用 Biacore 结合测定法评估了从人粪便中分离的 10 株乳杆菌和 3 株双歧杆菌对可溶性人结肠黏蛋白(sHCM)的黏附特性。选择了 3 株(乳杆菌 ME-522 株、Lact. gasseri ME-527 株和双歧杆菌 bifidum MCC1092 株)具有显著黏附能力的菌株。用唾液酸酶去除 sHCM 中的唾液酸后,观察到结合活性降低。然而,用硫酸酯酶去除硫酸酯残基后,ME-527 的黏附力下降;而其余两株的黏附力则增加。用氯化钡洗脱阻断硫酸盐残基后,三种益生菌的黏附能力显著下降。

结论

使用 Biacore 测定法开发了一种新的评价方法,观察对 sHCM 酸性残基的结合特性。结果表明,益生菌黏附和 sHCM 酸性残基之间存在很强的关系。

研究的意义和影响

这是第一个报告,使用 Biacore 结合测定法定量测量细菌与 sHCM 中酸性残基之间的结合的筛选方法;并为未来益生菌的选择提供了一种新方法。

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