Abstract
The objectives were to develop an assay method for detection and quantification, and determine kinetic parameters of β-D-fucosidase activity in soil. The optimal activities of this enzyme were at approximately pH 6.0 and 55 °C, respectively. The Km values, based on hydrolyzing p-nitrophenyl-β-D-fucopyranoside, in five soils ranged from 2.40 to 2.89 mM, and the Vmax values spanned from 206 to 1060 μmol p-nitrophenol released kg−1 soil h−1. At reaction temperatures from 10 to 40 °C, the mean temperature coefficient (Q10) and activation energy (Ea) ranged from 1.67 to 1.90, and 44.22 to 56.39 kJ mol−1, respectively. Employing a chromogenic p-nitrophenyl substrate, the developed method detects as little as 3.5 μmol p-nitrophenol released kg−1 soil h−1 (limit of detection or LOD), quantifies ≥ 10.7 μmol p-nitrophenol released kg−1 soil h−1 (limit of quantitation or LOQ), and had coefficients of variance (CV) of < 5.6%. β-D-fucosidases purified from some sources also hydrolyze β-D-galactoside, β-D-glucosides, and/or α-L-arabinosides. This method offers reproducibility and precision that are comparable with bench-scale assays, but also has the advantage of using a smaller amount of soil, reducing costs on reagents, supplies, labor, and waste management.
Published Version
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