New spatially explicit method for detecting extracellular protease activity in biofilms |
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Authors: | Francoeur S N Wetzel R G Neely R K |
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Affiliation: | Department of Biological Sciences, The University of Alabama, Tuscaloosa, Alabama 35487-0206, USA. steven_francoeur@hotmail.com |
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Abstract: | A novel method of detecting extracellular protease activity at biofilm-substratum interfaces was developed. This method utilizes fluorescent molecules bound to cellulose substrata with a lectin. Extracellular proteases degrade the lectin and release the fluorochrome into solution. This new technique and a standard dissolved-substrate assay detected similar responses of biofilm extracellular protease activity to experimental manipulation of N supply. Combination of this technique with confocal scanning laser microscopy allowed direct visualization of microspatial patterns of bacterial distribution and extracellular protease activity at the biofilm-substratum interface. |
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