Enzyme assay with fluoresceinamine labeled biopolymers
Template:BreadcrumbsSecondaryProduction
| Polysaccharide hydrolysis |
|---|
| Approach: enzyme assay |
| Context: incubation, lab |
| Spatial scale: low |
| Temporal scale: low; hours to days |
| Units: mol monomer L-1 h-1 |
| Community captured: heterotrophs |
| Co-measurements: cell abundance |
Method Overview
This enzyme assay measures polysaccharide hydrolysis by time course incubations with fluorescently labeled substrate (fluoresceinamine labeled biopolymers)[1].
The assay is performed in discrete mode on bulk, size-fract, single cell samples.
Scale of measurement
Typically performed on replicate samples collected at a sampling station at one point in time. Sampling effort is relatively high impairing resolution in space or time. Time course incubations last hours to days, further restricting temporal resolution.
Data generated
These measurements can provide hydrolysis rates per unit time.
Units & currency
Units are mol monomer L-1 h-1. The currency is carbon.
Sample size
Typical samples are < 1 L in volume.
Limitations
Rate measurements to estimate Vmax, yielding potential rates. Bottle effects are likely to occur.
Example Applications & Protocols
Classic examples
- Arnosti (1996) A new method for measuring polysaccharide hydrolysis rates in marine environments [1]
Recent applications
- Martinez-Garcia et al. (2012) Capturing Single Cell Genomes of Active Polysaccharide Degraders: An Unexpected Contribution of Verrucomicrobia [2]
References
- ↑ 1.0 1.1 Arnosti (1996). A new method for measuring polysaccharide hydrolysis rates in marine environments. Organic Geochemistry, 25(1-2), 105-115. https://doi.org/10.1016/S0146-6380(96)00112-X
- ↑ Martinez-Garcia et al. (2012). Capturing Single Cell Genomes of Active Polysaccharide Degraders: An Unexpected Contribution of Verrucomicrobia. PLoS ONE, 7(4), e35314. https://doi.org/10.1371/journal.pone.0035314