Schmid Jochen, Rühmann Broder, Sieber Volker, Romero-Jiménez Lorena, Sanjuán Juan, Pérez-Mendoza Daniel
Chair of Chemistry of Biogenic Resources, Technical University of Munich, Straubing, Germany.
Dpto. Microbiología del Suelo y Sistemas Simbióticos, Estación Experimental del Zaidín, CSIC, Granada, Spain.
Methods Mol Biol. 2018;1734:263-275. doi: 10.1007/978-1-4939-7604-1_21.
Bacterial exopolysaccharides (EPS) often confer a survival advantage by protecting the cell against abiotic and biotic stresses, including host defensive factors. They are also main components of the extracellular matrix involved in cell-cell recognition, surface adhesion and biofilm formation. Biosynthesis of a growing number of EPS has been reported to be regulated by the ubiquitous second messenger c-di-GMP, which promotes the transition to a biofilm mode of growth in an intimate association with the eukaryotic host. Here we describe a strategy based on the combination of an approach to artificially increase the intracellular level of c-di-GMP in virtually any gram-negative bacteria with a high throughput screening (HTS) for the identification of monosaccharide composition and carbohydrate fingerprinting of novel EPS, or modified variants, that can be involved in host-bacteria interactions.
细菌胞外多糖（EPS）通常通过保护细胞免受非生物和生物胁迫（包括宿主防御因子）而赋予生存优势。它们也是细胞外基质的主要成分，参与细胞间识别、表面粘附和生物膜形成。据报道，越来越多的EPS生物合成受普遍存在的第二信使环二鸟苷酸（c-di-GMP）调控，c-di-GMP与真核宿主密切相关，促进向生物膜生长模式的转变。在此，我们描述了一种策略，该策略基于将一种在几乎任何革兰氏阴性细菌中人工提高细胞内c-di-GMP水平的方法与高通量筛选（HTS）相结合，以鉴定新型EPS或修饰变体的单糖组成和碳水化合物指纹图谱，这些EPS或修饰变体可能参与宿主-细菌相互作用。
