Determination of the beta-glucosidase activity in different soils by pre capillary enzyme assay using capillary electrophoresis with laser-induced fluorescence detection.
Enzyme activities can provide indication for quantitative changes in soil organic matter (SOM). It is known that the activities of most enzymes increase as native SOM content reflecting larger microbial communities and stabilization of enzymes on humic materials. Beta-glucosidase (beta-Glu) activities have been frequently used as indicators of changes in quantity and quality of SOM. In this study we propose a simple and very sensitive method, which has lower limit of detection compared with classic spectrophotometric method with the aim of determinate the beta-Glu activity in soil samples using Fluorescein mono-beta-D-glucopyranoside (FMGlc) as a substrate. The fluorescein released by the enzymatic reaction was quantified by capillary electrophoresis-laser induced fluorescence (CE-LIF) method. The background electrolyte (BGE) consisted in 40 mM phosphate buffer, pH 6. The LOD and LOQ for fluorescein were 1.3 10(-7) mg mL(-1) and 6.4 10(-6) mg mL(-1), respectively. This work deals with the minimization of the mixture for the enzymatic reaction and with the optimization conditions of CE separation. To the best of our knowledge, this is the first time that an enzymatic activity was detected in soil using CE-LIF system.
Stege
Patricia W
PW
INQUISAL, Department of Chemistry, National University of San Luis, CONICET, Chacabuco y Pedernera, D5700BWS San Luis, Argentina. pwstege@unsl.edu.ar
Messina
Germán A
GA
Bianchi
Guillermo
G
Olsina
Roberto A
RA
D019075:Electrophoresis, Capillary / Q000379:methods*; D057075:Enzyme Assays / Q000379:methods*; D019793:Fluorescein / Q000737:chemistry; D005453:Fluorescence; D006863:Hydrogen-Ion Concentration; D007834:Lasers; D016014:Linear Models; D012680:Sensitivity and Specificity; D012987:Soil / Q000032:analysis*; D013053:Spectrophotometry / Q000379:methods; D014882:Water-Electrolyte Balance; D001617:beta-Glucosidase / Q000032:analysis*
D012987:Soil; D001617:beta-Glucosidase; D019793:Fluorescein