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Fluorometric Determination of the Activity of β-Glucosidase and Other Extracellular Hydrolytic Enzymes

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Methods to Study Litter Decomposition

Abstract

Cleavage of macromolecules by extracellular enzymes is a crucial process in the microbial decomposition of plant litter. Potential activities of cellulose-degrading and many other hydrolytic extracellular enzymes can be precisely measured by means of fluorogenic model substrates, most commonly 4-methylumbelliferone (MUF) linked to compounds such as glucose, phosphate, or amino acids. The bonds between these molecules are cleaved by the enzymes in a way similar to the natural oligomeric or polymeric substances. This chapter describes a method for determining β-glucosidase activity in decomposing plant litter as an example of a broad range of extracellular hydrolytic enzymes that can be quantified by this approach. Plant litter is homogenized and a small aliquot of the homogenate is mixed with the fluorogenic MUF compound. Fluorescence is determined after 5 min and again, for β-glucosidase, after an incubation period of 60 min. The fluorescent MUF released by hydrolysis indicates the level of extracellular enzyme activity in the sample. Potential activities of β-glucosidase in decomposing leaves and wood from aquatic and soil environments determined by this method vary by up to three orders of magnitude, ranging from 1 to 4300 μmol per gram of organic matter per hour.

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Correspondence to Jürgen Marxsen .

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Hendel, B., Marxsen, J. (2020). Fluorometric Determination of the Activity of β-Glucosidase and Other Extracellular Hydrolytic Enzymes. In: Bärlocher, F., Gessner, M., Graça, M. (eds) Methods to Study Litter Decomposition. Springer, Cham. https://doi.org/10.1007/978-3-030-30515-4_44

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