Journal
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
Volume -, Issue 80, Pages -Publisher
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/50399
Keywords
Environmental Sciences; Issue 80; Environmental Monitoring; Ecological and Environmental Processes; Environmental Microbiology; Ecology; extracellular enzymes; freshwater microbiology; soil microbiology; microbial activity; enzyme activity
Categories
Funding
- United States Department of Agriculture [58-6408-1-595]
- National Science Foundation [1049911]
- Direct For Biological Sciences
- Division Of Environmental Biology [1049911] Funding Source: National Science Foundation
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Much of the nutrient cycling and carbon processing in natural environments occurs through the activity of extracellular enzymes released by microorganisms. Thus, measurement of the activity of these extracellular enzymes can give insights into the rates of ecosystem level processes, such as organic matter decomposition or nitrogen and phosphorus mineralization. Assays of extracellular enzyme activity in environmental samples typically involve exposing the samples to artificial colorimetric or fluorometric substrates and tracking the rate of substrate hydrolysis. Here we describe microplate based methods for these procedures that allow the analysis of large numbers of samples within a short time frame. Samples are allowed to react with artificial substrates within 96-well microplates or deep well microplate blocks, and enzyme activity is subsequently determined by absorption or fluorescence of the resulting end product using a typical microplate reader or fluorometer. Such high throughput procedures not only facilitate comparisons between spatially separate sites or ecosystems, but also substantially reduce the cost of such assays by reducing overall reagent volumes needed per sample.
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