Inhibition of Streptococcus mutans Biofilm Formation by Streptococcus salivarius FruA
Inhibition of Streptococcus mutans Biofilm Formation by Streptococcus salivarius FruA
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DOI：
10.1128/aem.02066-10
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发表时间：
2011-03-01
影响因子：
4.4
通讯作者：
Senpuku, Hidenobu
中科院分区：
文献类型：
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作者：
Ogawa, Ayako;Furukawa, Soichi;Senpuku, Hidenobu
口腔微生物区系由许多有益的细菌组成，这些细菌与健康状况有关，并控制口腔疾病的进展。口腔链球菌与病原菌的协同作用在口腔内牙科生物膜的形成过程中起着重要作用。为确定口腔链球菌在多种生物膜形成中的作用以及在生物膜形成中的作用，采用离子交换和凝胶过滤层析从唾液链球菌ATCC 9759和HT9R培养上清液中分离纯化了抑制变形链球菌生物被膜形成的活性物质。进行基质辅助激光解吸电离飞行时间(MALDI-TOF)质谱分析，并与数据库进行比较。测定了唾液酸链霉菌HT9R基因组序列，并用于鉴定抑制的候选蛋白。抑制生物被膜的候选酶为唾液链霉菌果糖基转移酶(FTF)和外β-D-果糖苷酶(FruA)。在蔗糖存在的情况下，抑制物的活性增强，并且抑制作用依赖于生物被膜形成试验中蔗糖的浓度。纯化的和商品化的黑曲霉果肉(与唾液葡萄球菌HT9R果肉的氨基酸序列同源性分别为31.6%和59.6%)完全抑制变形链球菌GS-5生物膜在唾液包被的聚苯乙烯和羟基磷灰石表面的形成。抑制是通过减少多糖的产生而引起的，而多糖的产生依赖于蔗糖的消化而不是果聚糖的消化。这些数据表明，唾液链霉菌产生大量的FruA，FruA本身可能在口腔中依赖蔗糖的生物膜形成的多物种微生物相互作用中发挥重要作用。
The oral microbial flora consists of many beneficial species of bacteria that are associated with a healthy condition and control the progression of oral disease. Cooperative interactions between oral streptococci and the pathogens play important roles in the development of dental biofilms in the oral cavity. To determine the roles of oral streptococci in multispecies biofilm development and the effects of the streptococci in biofilm formation, the active substances inhibiting Streptococcus mutans biofilm formation were purified from Streptococcus salivarius ATCC 9759 and HT9R culture supernatants using ion exchange and gel filtration chromatography. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry analysis was performed, and the results were compared to databases. The S. salivarius HT9R genome sequence was determined and used to indentify candidate proteins for inhibition. The candidates inhibiting biofilms were identified as S. salivarius fructosyltransferase (FTF) and exo-beta-D-fructosidase (FruA). The activity of the inhibitors was elevated in the presence of sucrose, and the inhibitory effects were dependent on the sucrose concentration in the biofilm formation assay medium. Purified and commercial FruA from Aspergillus niger (31.6% identity and 59.6% similarity to the amino acid sequence of FruA from S. salivarius HT9R) completely inhibited S. mutans GS-5 biofilm formation on saliva-coated polystyrene and hydroxyapatite surfaces. Inhibition was induced by decreasing polysaccharide production, which is dependent on sucrose digestion rather than fructan digestion. The data indicate that S. salivarius produces large quantities of FruA and that FruA alone may play an important role in multispecies microbial interactions for sucrose-dependent biofilm formation in the oral cavity.