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Enzyme assay with RBB labeled polysaccharides: Difference between revisions

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Created page with "{{BreadcrumbsSecondaryProduction}} * Page authors: PRIMO * Responsible curator: Hagen Buck-Wiese ---- __TOC__ <div class="model-box"> {| class="model-ib" style="float:right; margin-left:1em; margin-bottom:1em;" ! Polysaccharide hydrolysis |- | '''Approach:''' enzyme assay |- | '''Context:''' incubation, lab |- | '''Spatial scale:''' low |- | '''Temporal scale:''' low; hours to days |- | '''Units:'''..."
 
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== Method Overview ==
== Method Overview ==


Time course incubation with RBB labeled polysaccharides<ref name="0038071792900748">Wirth & Wolf (1992). Micro-plate colourimetric assay for Endo -acting cellulase, xylanase, chitinase, 1,3-β-glucanase and amylase extracted from forest soil horizons. ''Soil Biology and Biochemistry'', 24(6), 511-519. https://doi.org/10.1016/0038-0717(92)90074-8</ref>.
This enzyme assay measures polysaccharide hydrolysis by time course incubation with RBB labeled polysaccharides<ref name="Wirth1992">Wirth & Wolf (1992). Micro-plate colourimetric assay for Endo -acting cellulase, xylanase, chitinase, 1,3-β-glucanase and amylase extracted from forest soil horizons. ''Soil Biology and Biochemistry'', 24(6), 511-519. https://doi.org/10.1016/0038-0717(92)90074-8</ref>.


Applicable variants include: bulk.
The assay is performed in discrete mode on bulk samples.


=== Scale of measurement ===
=== Scale of measurement ===


Spatial resolution is low (horizontal) and low (vertical). Temporal resolution is low, with a time integration of hours to days.
<!-- TODO: describe spatial and temporal scale of this method -->


=== Data generated ===
=== Data generated ===


Yields rates in units of relative OD units.
<!-- TODO: describe what data this method generates and what it represents -->


=== Units & currency ===
=== Units & currency ===
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=== Sample size ===
=== Sample size ===


< L.
Typical samples are < 1 L in volume.


=== Repositories & databases ===
=== Repositories & databases ===
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== Limitations ==
== Limitations ==


rate measurements at max (Vmax). bottle effects, potential rates.
These rate measurements report potential rates (Vmax). Bottle effects can occur due to the length of the required incubation period.


== Example Applications & Protocols ==
== Example Applications & Protocols ==


=== Classic examples ===
=== Classic examples ===
* Wirth & Wolf (1992) ''Micro-plate colourimetric assay for Endo -acting cellulase, xylanase, chitinase, 1,3-β-glucanase and amylase extracted from forest soil horizons'' <ref name="0038071792900748" />
* Wirth & Wolf (1992) ''Micro-plate colourimetric assay for Endo -acting cellulase, xylanase, chitinase, 1,3-β-glucanase and amylase extracted from forest soil horizons'' <ref name="Wirth1992" />


=== Recent applications ===
=== Recent applications ===
* Zohary et al. (2000) ''The role of nutrients in decomposition of a thecate dinoflagellate'' <ref name="lo20004510123">Zohary et al. (2000). The role of nutrients in decomposition of a thecate dinoflagellate. ''Limnology and Oceanography'', 45(1), 123-130. https://doi.org/10.4319/lo.2000.45.1.0123</ref>
* Zohary et al. (2000) ''The role of nutrients in decomposition of a thecate dinoflagellate'' <ref name="Zohary2000">Zohary et al. (2000). The role of nutrients in decomposition of a thecate dinoflagellate. ''Limnology and Oceanography'', 45(1), 123-130. https://doi.org/10.4319/lo.2000.45.1.0123</ref>


=== Common calculations/conversions ===
=== Common calculations/conversions ===

Latest revision as of 11:33, 11 May 2026

Template:BreadcrumbsSecondaryProduction

Polysaccharide hydrolysis
Approach: enzyme assay
Context: incubation, lab
Spatial scale: low
Temporal scale: low; hours to days
Units: relative OD units
Community captured: heterotrophs
Co-measurements: cell abundance

Method Overview

This enzyme assay measures polysaccharide hydrolysis by time course incubation with RBB labeled polysaccharides[1].

The assay is performed in discrete mode on bulk samples.

Scale of measurement

Data generated

Units & currency

Units are relative OD units. The currency is carbon.

Sample size

Typical samples are < 1 L in volume.

Repositories & databases

Limitations

These rate measurements report potential rates (Vmax). Bottle effects can occur due to the length of the required incubation period.

Example Applications & Protocols

Classic examples

  • Wirth & Wolf (1992) Micro-plate colourimetric assay for Endo -acting cellulase, xylanase, chitinase, 1,3-β-glucanase and amylase extracted from forest soil horizons [1]

Recent applications

  • Zohary et al. (2000) The role of nutrients in decomposition of a thecate dinoflagellate [2]

Common calculations/conversions

References

  1. ↑ 1.0 1.1 Wirth & Wolf (1992). Micro-plate colourimetric assay for Endo -acting cellulase, xylanase, chitinase, 1,3-β-glucanase and amylase extracted from forest soil horizons. Soil Biology and Biochemistry, 24(6), 511-519. https://doi.org/10.1016/0038-0717(92)90074-8
  2. ↑ Zohary et al. (2000). The role of nutrients in decomposition of a thecate dinoflagellate. Limnology and Oceanography, 45(1), 123-130. https://doi.org/10.4319/lo.2000.45.1.0123
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