Micro-plate colourimetric assay for endo-acting cellulase, xylanase, chitinase, 1,3-beta-glucanase and amylase extracted from forest soil horizons
1992
Wirth, S.J. | Wolf, G.A.
A new approach, based on the application of soluble, dye-labelled and acid-precipitable polysaccharide derivatives, is introduced for the sensitive assay of polysaccharide endo-hydrolases extracted from soil. An extraction procedure involving sodium acetate acetic acid buffer (pH 5, 0.5 m: 5 ml g-1 soil) and an assay system adapted to microtitre plates were developed for routine determinations of endo-acting cellulase, xylanase, chitinase, 1.3-beta-glucanase and amylase activities. An Acid Brown Earth under a mature beech forest (Fagus sylvatica L., humusform: typical Moder) was studied with respect to distinct, clearly-developed soil horizons (L, F, H, Ahh, Aeh). Enzymes purified by (NH4)2SO4-precipitation and dialysis were assayed for pH and temperature activity profiles. pH optima were determined in the range of 4.5-5.5, temperature optima were in the range of 40-55 degrees C, revealing stable and fairly similar characteristics of these enzymes in the horizons under study. In routine investigations, highest enzyme activities were determined in the L and F horizons. Significantly decreasing activities with increasing soil depth and decreasing organic matter content were determined in the H. Ahh and Aeh horizons, respective]y Cellulase, xylanase and chitinase activities were highly correlated with total-C content of the soil horizons under study (0.871 less than or equal to r2 less than or equal to 0.954).
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