RRC ID 35649
著者 Nguyen MH, Ojima Y, Sakka M, Sakka K, Taya M.
タイトル Probing of exopolysaccharides with green fluorescence protein-labeled carbohydrate-binding module in Escherichia coli biofilms and flocs induced by bcsB overexpression.
ジャーナル 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.
巻・号 118(4)
ページ 400-5
公開日 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
引用数 12
WOS 分野 FOOD SCIENCE & TECHNOLOGY BIOTECHNOLOGY & APPLIED MICROBIOLOGY
リソース情報
原核生物(大腸菌) NA