Wickström C, Svensäter G
Department of Oral Biology, Faculty of Odontology, Malmö University, Malmö, Sweden.
Oral Microbiol Immunol. 2008 Jun;23(3):177-82. doi: 10.1111/j.1399-302X.2007.00407.x.
Model systems with oral bacteria from dental plaque have demonstrated that the utilization of complex glycoproteins as a food source cannot be undertaken by single species but requires concerted degradation by a multi-species consortium, with each member contributing one or a few hydrolytic enzymes. Unlike previous studies, the aim of the present investigation was to explore the ability of fresh dental plaque to degrade salivary mucin, MUC5B, isolated by methods designed to retain intact the natural polymeric structure and physiological conformation, in an attempt to mimic the naturally occurring interaction between the oral microbiota and salivary mucins.
Human salivary MUC5B was isolated from whole saliva by density-gradient centrifugation and incubated with freshly isolated supragingival dental plaque with samples subjected to fluorescent staining for viability and metabolic activity. In addition, the degradation of MUC5B oligosaccharide side chains was studied using a lectin assay, recognizing three different carbohydrate epitopes commonly found on mucin oligosaccharide side chains.
The addition of purified salivary MUC5B elicited a strong metabolic response from the biofilm cells, whereas individual strains of Streptococcus oralis and Streptococcus gordonii isolated from the same plaque were not able to utilize the MUC5B. The degradation of terminal saccharide moieties on the MUC5B was demonstrated by a marked decrease in both sialic acid and fucose reactivity.
These results have shown that dental plaque is capable of utilizing human salivary MUC5B as a nutrient source, a process possibly requiring the synergistic degradation of the molecule by a consortium of oral bacteria in the plaque community.
含有牙菌斑中口腔细菌的模型系统已表明,单一物种无法利用复合糖蛋白作为食物来源,而是需要多种物种组成的菌群协同降解,每个成员贡献一种或几种水解酶。与以往研究不同的是,本研究的目的是探索新鲜牙菌斑降解唾液黏蛋白MUC5B的能力,该黏蛋白是通过旨在保持天然聚合物结构和生理构象完整的方法分离得到的,旨在模拟口腔微生物群与唾液黏蛋白之间自然发生的相互作用。
通过密度梯度离心从全唾液中分离出人唾液MUC5B,并与新鲜分离的龈上牙菌斑一起孵育,对样本进行荧光染色以检测活力和代谢活性。此外,使用凝集素测定法研究MUC5B寡糖侧链的降解情况,该方法可识别黏蛋白寡糖侧链上常见的三种不同碳水化合物表位。
添加纯化的唾液MUC5B引发了生物膜细胞强烈的代谢反应,而从同一菌斑中分离出的口腔链球菌和戈登链球菌的单个菌株无法利用MUC5B。唾液酸和岩藻糖反应性均显著降低,证明了MUC5B末端糖基部分的降解。
这些结果表明,牙菌斑能够将人唾液MUC5B作为营养源利用,这一过程可能需要菌斑群落中的口腔细菌菌群对该分子进行协同降解。