Rapid isolation and high-throughput determination of cellulase and laminarinase activity in soils.
A new method for extracting soil enzymes is described and a microplate method for assaying soil beta-1,4-glucanases (cellulases) and beta-1,3-glucanases (laminarinases). Soil samples were mechanically disrupted to produce crude enzyme extracts, and diluted preps incubated in microplates containing either carboxymethyl cellulose (CMC) to determine cellulase activity or laminarin substrate to determine laminarinase activity. The resulting glucose was measured using the fluorometric Amplex Red glucose assay. The method was reproducible, could be completed in 1 day and measured twice as much enzyme activity than the standard passive soil enzyme extraction procedure. The method described herein facilitates the development of high-throughput soil multiplex enzymatic assays from several soil samples at one time, and is well suited to the study of functional microbial ecology.
Vancov
T
T
NSW Department of Primary Industries, Wollongbar Agricultural Institute, 1243 Bruxner Highway, Wollongbar, 2477 New South Wales, Australia. tony.vancov@dpi.nsw.gov.au
Keen
B
B
D002266:Carboxymethylcellulose Sodium / Q000378:metabolism; D002480:Cellulase / Q000302:isolation & purification / Q000378:metabolism*; D044602:Cellulases / Q000302:isolation & purification / Q000378:metabolism*; D005936:Glucans; D005947:Glucose / Q000032:analysis; D011134:Polysaccharides / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012987:Soil / Q000032:analysis*
D005936:Glucans; D011134:Polysaccharides; D012987:Soil; C008247:laminaran; D044602:Cellulases; D002480:Cellulase; D005947:Glucose; D002266:Carboxymethylcellulose Sodium