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4-MUF Phosphate

FIGURE 3.5 Enzyme assays using different nonfluorescent substrates and measuring highly fluorescent products after a short-term incubation.

opportunity to detect enzyme activities of small sample sizes (e.g., microaggregates and rhizosphere samples) and/or low activity (e.g., samples of subsoil, peat, and soil solutions). The development of an improved automated assay using a 96-well microplate reader provides a convenient system for processing large numbers of different samples; it also allows the activity rates to be measured for a range of enzymes in a number of soils with an appropriate number of replicates (Vepsalainen et al., 2001). Alternatively, a multiple-substrate approach is available, which is based on the simultaneous measurement of seven enzyme activities estimated after incubation, separation of the nonhydrolyzed 4-MUF and 7-AMC substrates by HPLC, and quantification by UV-absorption at 320 nm (Stemmer, 2004).

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