Reference - Detail
| RRC ID | 35649 |
|---|---|
| Author | Nguyen MH, Ojima Y, Sakka M, Sakka K, Taya M. |
| Title | Probing of exopolysaccharides with green fluorescence protein-labeled carbohydrate-binding module in Escherichia coli biofilms and flocs induced by bcsB overexpression. |
| Journal | J Biosci Bioeng |
| Abstract |
Polysaccharides are major structural constituents to develop the three-dimensional architecture of Escherichia coli biofilms. In this study, confocal laser scanning microscopy was applied in combination with a fluorescent probe to analyze the location and arrangement of exopolysaccharide (EPSh) in microcolonies of E. coli K-12 derived strains, formed as biofilms on solid surfaces and flocs in the liquid phase. For this purpose, a novel fluorescent probe was constructed by conjugating a carbohydrate-binding module 3, from Paenibacillus curdlanolyticus, with the green fluorescence protein (GFP-CBM3). The GFP-CBM3 fused protein exhibited strong affinity to microcrystalline cellulose. Moreover, GFP-CBM3 specifically bound to cell-dense microcolonies in the E. coli biofilms, and to their flocs induced by bcsB overexpression. Therefore, the fused protein presents as a novel marker for EPSh produced by E. coli cells. Overexpression of bcsB was associated with abundant EPSh production and enhanced E. coli biofilm formation, which was similarly detectable by GFP-CBM3 probing. |
| Volume | 118(4) |
| Pages | 400-5 |
| Published | 2014-10-1 |
| DOI | 10.1016/j.jbiosc.2014.03.005 |
| PII | S1389-1723(14)00104-2 |
| PMID | 24746734 |
| MeSH | Bacterial Proteins / genetics Bacterial Proteins / metabolism* Biofilms Cellulose / chemistry Escherichia coli K12 / genetics Escherichia coli K12 / metabolism* Fluorescence Fluorescent Dyes / chemistry* Gene Expression Green Fluorescent Proteins / genetics Green Fluorescent Proteins / metabolism* Microscopy, Confocal Paenibacillus / chemistry Polysaccharides, Bacterial / biosynthesis* Receptors, Cell Surface / genetics Receptors, Cell Surface / metabolism* Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / metabolism |
| IF | 2.366 |
| Times Cited | 12 |
| WOS Category | FOOD SCIENCE & TECHNOLOGY BIOTECHNOLOGY & APPLIED MICROBIOLOGY |
| Resource | |
| Prokaryotes E. coli | NA |