Núcleo Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, Brazil; Faculdade de Medicina, Laboratório de Micologia, Universidade Federal de Rio Grande, Rio Grande, Brazil.
J Appl Microbiol. 2013 Nov;115(5):1222-30. doi: 10.1111/jam.12318. Epub 2013 Sep 3.
The aim of the present work was to study the in vitro effect of native and recombinant Bauhinia variegata var. variegata lectins in inhibiting early adhesion of Streptococcus mutans, Streptococcus sanguis and Streptococcus sobrinus to experimentally acquired pellicle.
Native lectin from B. variegata (BVL) was purified by affinity chromatography of extract of seeds. The recombinant lectin (rBVL-I) was expressed in E. coli strain BL21 (DE3) from a genomic clone encoding the mature B. variegata lectin gene using the vector pAE-bvlI. Recombinant protein deposited in inclusion bodies was solubilized and subsequently purified by affinity chromatography. The rBVL-I was compared to BVL for agglutination of erythrocytes and initial adherence of oral bacteria on a saliva-coated surface. The results revealed that rBVL-I acts similarly to BVL for agglutination of erythrocytes. Both lectins showed adhesion inhibition effect on Step. sanguis, Step. mutans and Step. sobrinus.
We report, for the first time, the inhibition of early adhesion of oral bacteria by a recombinant lectin.
Our results support the proposed biotechnological application of lectins in a strategy to reduce development of dental caries by inhibiting the initial adhesion and biofilm formation.
本研究旨在研究天然和重组猪豆花凝集素对变异猪豆花凝集素体外抑制变形链球菌、血链球菌和远缘链球菌早期黏附实验获得的获得性膜的影响。
从种子提取物中通过亲和层析法纯化天然猪豆花凝集素(BVL)。使用载体 pAE-bvlI 从编码成熟猪豆花凝集素基因的基因组克隆中在大肠杆菌菌株 BL21(DE3)中表达重组凝集素(rBVL-I)。重组蛋白以包涵体形式沉淀,然后通过亲和层析进行纯化。比较 rBVL-I 与 BVL 对红细胞的凝集作用以及口腔细菌在唾液包被表面的初始黏附作用。结果表明,rBVL-I 与 BVL 对红细胞的凝集作用相似。两种凝集素均对血链球菌、变形链球菌和远缘链球菌表现出黏附抑制作用。
我们首次报道了重组凝集素抑制口腔细菌早期黏附的作用。
我们的结果支持了凝集素在减少龋齿发展的策略中的生物技术应用,通过抑制初始黏附和生物膜形成。