title: New spatially explicit method for detecting extracellular protease activity in biofilms.
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.
authors.affiliation: Department of Biological Sciences, The University of Alabama, Tuscaloosa, Alabama 35487-0206, USA. steven_francoeur@hotmail.com
authors.forename: S N
authors.initials: SN
authors.lastname: Francoeur
authors.forename: R G
authors.initials: RG
authors.lastname: Wetzel
authors.forename: R K
authors.initials: RK
authors.lastname: Neely
mesh_terms: D001419:Bacteria / Q000201:enzymology* / Q000254:growth & development; D018441:Biofilms / Q000254:growth & development*; D010450:Endopeptidases / Q000378:metabolism*; D019793:Fluorescein / Q000378:metabolism; D008851:Micropore Filters; D018613:Microscopy, Confocal / Q000379:methods; D014909:Wheat Germ Agglutinins / Q000378:metabolism
chemical_list: D014909:Wheat Germ Agglutinins; D010450:Endopeptidases; D019793:Fluorescein