Difference between revisions of "Team:TU Darmstadt/Demonstrate"

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<figcaption width="80%> <b>Figure 1. Activity assay of NodC.</b> The NodC (40 ng) was titrated in 1X glycosyltransferase reaction buffer the presence of 100&nbsp;μM of UDP-<i>N</i>-acetylglcosamine and 10&nbsp;mM <i>N</i>-acetylglucosamine (GlcNAc) as an acceptor substrate. The reaction was performed as described before and the luminescence was measured after 1 hour of incubation with a <i>Tecan200 Infinite Pro</i> plate reader. Each point is an average of two experiments, and the error bars represent the standard deviations. RLU = relative light units.</center>
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<figcaption width="80%> <b>Figure 1. Activity assay of NodC.</b> The NodC (40 ng) was titrated in 1X glycosyltransferase reaction buffer the presence of 100&nbsp;μM of UDP-<i>N</i>-acetylglcosamine and 10&nbsp;mM <i>N</i>-acetylglucosamine (GlcNAc) as an acceptor substrate. The reaction was performed as described before and the luminescence was measured after 1 hour of incubation with a <i>Tecan200 Infinite Pro</i> plate reader. Each point is an average of two experiments, and the error bars represent the standard deviations. RLU = relative light units.
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Revision as of 21:16, 29 October 2017

MainPage

Proof of Concept

Here we want to give you a short tour about what we achieved in the project. We show our results and give a view on the resulting product, a enzyme sensing hydrogel.

Chitin Synthase NodC

The functionality of the NodC enzyme was verified by performing the UDP-Glo™ Glycosyltransferase Assay. The evaluation of the assay shows that the NodC enzyme converts the UDP-GlcNAc to free UPD and a growing oligo-GlcNAc-chain. The free UDP is converted to ATP, which acts as a substrate for a luciferase reaction and creates luminescence. So the assay and the increasing luminescence depending on increasing enzyme concentrations shows that the NodC enzyme can create chitin oligomers.