Open AccessNew Spatially Explicit Method for Detecting Extracellular Protease Activity in Biofilms
Author(s) -
Steven N. Francoeur,
Robert G. Wetzel,
Robert K. Neely
Publication year - 2001
Publication title -
applied and environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.552
H-Index - 324
eISSN - 1070-6291
pISSN - 0099-2240
DOI - 10.1128/aem.67.9.4329-4334.2001
Subject(s) - biofilm , extracellular , protease , proteases , extracellular matrix , confocal microscopy , biophysics , lectin , extracellular polymeric substance , cellulose , substrate (aquarium) , microbiology and biotechnology , chemistry , biochemistry , biology , bacteria , enzyme , ecology , genetics
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.