Evaluation of fluorescently labeled lectins for noninvasive localization of extracellular polymeric substances in Sphingomonas biofilms

被引:77
作者
Johnsen, AR
Hausner, M
Schnell, A
Wuertz, S
机构
[1] Tech Univ Munich, Inst Water Qual Control & Waste Mangement, D-85748 Garching, Germany
[2] Natl Environm Res Inst, Dept Microbial Ecol & Biotechnol, DK-4000 Roskilde, Denmark
关键词
D O I
10.1128/AEM.66.8.3487-3491.2000
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Three strains of Sphingomonas were grown as biofilms and tested for binding of five fluorescently labeled lectins (Con A-type IV-TRITC or -Cy5, Pha-E-TRITC, PNA-TRITC, UEA 1-TRITC, and WGA-Texas red). Only ConA and WGA were significantly bound by the biofilms. Binding of the live lectins to artificial biofilms made of the commercially available Sphinogomonas extracellular polysaccharides was similar to binding to living biofilms, Staining of the living and artificial biofilms by ConA might be explained as binding of the lectin to the terminal mannosyl and terminal glucosyl residues in the polysaccharides secreted by Sphingomonas as well as to the terminal mannosyl residue in glycosphingolipids. Staining of the biofilms by WGA could only be explained as binding to the Sphingomonas glycosphingolipid membrane, binding to the cell wall, or nonspecific binding. Glycoconjugation of ConA and WGA with the target sugars glucose and N-acetylglucosamine, respectively, was used as a method for evaluation of the specificity of the lectin; towards Sphingomonas biofilms and Sphingomonas polysaccharides. Our results show that the binding of lectins to biofilms does not necessarily prove the presence of specific target sugars in the extracellular polymeric substances (EPS) in biofilms, The lectins may bind to non-EPS targets or adhere nonspecifically to components of the biofilm matrix.
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页码:3487 / 3491
页数:5
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